Skip to main content
NCBI home page
Carcinogenesis logoLink to Carcinogenesis
. 2025 Jan 1;46(1):bgae081. doi: 10.1093/carcin/bgae081

Novel inhibitory effect of Omega-3 fatty acids regulating pancreatic cancer progression

María I Garay 1,2, Tamara Mazo 3,4, Victoria Ferrero 5,6, Nelso N Barotto 7, Clarisa Lagares 8, María F Granton 9, María J Moreira-Espinoza 10, David C Cremonezzi 11, Andrea Comba 12, Mabel N Brunotto 13,14, Ezequiel J Tolosa 15, Martín E Fernandez-Zapico 16, María E Pasqualini 17,18,✉
PMCID: PMC12097994  PMID: 39742417

Abstract

Pancreatic cancer is a devastating malignancy in great need of new and more effective treatment approaches. In recent years, studies have indicated that nutritional interventions, particularly nutraceuticals, may provide novel avenues to modulate cancer progression. Here, our study characterizes the impact of ω-3 polyunsaturated fatty acids, eicosapentaenoic acid, and docosahexaenoic acid, as a nutraceutical intervention in pancreatic cancer using a genetically engineered mouse model driven by KrasG12D and Trp53R172H. This model closely resembles human pancreatic carcinogenesis, offering a disease relevant platform for translational research. Our findings showed that ω-3 polyunsaturated fatty acids intervention (using a diet supplemented with 6% cod liver oil) significantly reduced tumor volume as well as lung and liver metastasis and a trend toward improved survival rate compared with control treated mice. This antitumoral effect was accompanied by distinct changes in tumor membrane fatty acid profile and eicosanoids release. Furthermore, the eicosapentaenoic acid and docosahexaenoic acid intervention also reduced malignant histological parameters and induced apoptosis without affecting cell proliferation. Of note is the significant reduction in tumor fibrosis that was associated with decreased levels of Sonic Hedgehog, a major ligand controlling this cellular compartment in pancreatic cancer. All together our results demonstrate the impact of eicosapentaenoic acid and docosahexaenoic acid as antitumor regulators in pancreatic cancer, suggesting potential for ω-3 polyunsaturated fatty acids as a possible antitumoral dietary intervention. This research opens new avenues for integrating nutraceutical strategies in pancreatic cancer management.

Keywords: pancreatic cancer, ω-3 polyunsaturated fatty acids, nutraceutical intervention, Sonic Hedgehog

Our ω-3 fatty acids-based intervention impairs pancreatic cancer progression with a significant impact in the tumor microenvironment. Thus, supporting the translation value of this strategy for future clinical studies aimed at modulating pancreatic carcinogenesis.

Graphical Abstract

Graphical Abstract.

Graphical Abstract

Open in a new tab

Introduction

Despite considerable progress in research and medical technology, pancreatic cancer continues to have high mortality rates (1). This can be primarily attributed to the aggressive nature of the tumor, the absence of early symptoms, and its resistance to standard of care chemotherapeutic agents (2). Novel and more efficacious therapies are needed to improve the outcome of pancreatic cancer patients. In this context, some studies have indicated that dietary interventions might improve the effectiveness of therapy for pancreatic cancer (3). However, current research in this area remains limited and mostly derived from preclinical models that do not accurately reflect the intricate pathobiology of human pancreatic cancer.

Thus, to investigate these potential benefits of dietary therapeutic intervention, we utilized the KPC mouse (KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre) as a preclinical model (4). This genetically engineered model stands out for its remarkable similarity to human pancreatic cancer, making it an ideal platform for translational research. It maintains a fully functional immune response and accurately mimics the dense stromal reaction characteristic of human condition, providing a suitable model for the testing new therapeutic strategies (5,6), thus enhancing the translational potential of our findings. As interventions, we used cod liver oil high in ω-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) that represent a promising approach as preventive agents and a potential nutraceutics. These lipids have demonstrated strong anti-cancer properties (7,8) and exhibit high bioavailability to pancreatic cells, with research indicating that the pancreas can accumulate these fatty acids more efficiently than other tissues (9,10). This enhanced absorption in pancreatic tissue potentially increases their efficacy for intervention for pancreatic cancer. Here, we demonstrated a novel inhibitory of these ω-3 fatty acids blocking primary tumor growth and metastasis with a significant impact in the pancreatic tumor microenvironment and importantly, resulted in a trend to improve the survival rate. Our finding demonstrated the therapeutic value of high ω-3 diet (Hω-3) as new strategy to treat pancreatic cancer that could be integrated into standard treatment regimens, potentially enhancing their effectiveness of conventional therapies while offering the advantages of nutraceutical interventions. These benefits include potentially fewer side effects compared with traditional chemotherapies, the ability to complement existing treatments, and the possibility of long-term use for both cancer prevention and management.

Material and methods

KPC mice

We used the KPC mouse model of pancreatic cancer which incorporates, through Cre-Lox technology, the conditional activation of mutant endogenous alleles of the Kras and Trp53 genes (4). Specifically, an activating point mutation (G12D) in Kras and a dominant negative mutation in Trp53 (R172H) are conditionally activated in the mouse pancreas by breeding LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre (KPC) mice that express Cre recombinase under the expression of the pancreas-specific Pdx-1 promoter. For the genotyping, mouse ear DNA extraction was performed using the DNA easy kit (Qiagen, Hilden, Germany) and Taq Polymerase kit (Promega, Madison, WI, USA) following the manufacturer’s instructions. The primers used were the following: Kras: 5´-agctagccaccatggcttgagtaagtctgca-3´ (forward), Kras: 3´-cctttacaagcgcacgcagactgtaga-5´ (reverse); p48-Cre: 5´-agatgttcgcgattatcttc-3´ (forward), p48-Cre: 3´-agctacaccagagacgg-5´ (reverse); and Trp53: 5´-aaggggtatgagggacaagg-3´ (forward), Trp53: 3´-gaagacagaaaaggggaggg-5´ (reverse). The PCR cycling conditions were as follows: 95º for 2 minutes, 95º for 30 seconds, melting for 45 seconds (Kras: 65.8º; p48-Cre: 52.5º; p53: 57.7º), and 72º for 1 minutes for 35 cycles, followed by 72° for 10 minutes. The amplified products by PCR were run on a 1.2% agarose gel with molecular weight marker and the amplified products were visualized under UV trans-illuminator (11).

Diets and feeding protocol

Animals received two different diets sufficient in essential fatty acids throughout the length of the experiment, the Control, and Hω-3 diets; female and male mice were randomly divided into the experimental groups (n = 8–12/group). Mice were housed in groups of two to four in polycarbonate cages. Food and water were provided ad libitum. Animals were maintained at a constant temperature of 20°C and subjected to a 12-hour light/12-hour darkness cycle. Both base diets were composed of: 6% of tested oil, 16% casein, 34% sucrose, 39% corn starch, 2% fiber, 2.5% mineral salt mixture, and 0.5% vitamin mixture. The Control diet was added with 6% of Corn oil and the Hω-3 diet was added with 6% of fish oil (Supplementary Table 1). Oils identified by each group: CO (control, corn oil, Zea mays, high ω-6, and low ω-3 was purchased from ARCOR S.A., Cordoba, Argentina) and Fish oil: FO (cod liver oil, Hω-3) was purchased from Pura Química S.R.L., Córdoba, Argentina according to previous experiments (12). Corn oil contained 54% of linoleic acid (LA, 18:2 ω6), 0.6% of alpha linolenic acid (ALA, 18:3 ω3), 21% of oleic acid (OA, 18:1 ω9), and 15% of palmitic acid (PA, 16:0). Fish oil contained 14% of EPA (20:5 ω3), 32% of DHA (22:6 ω3), 1.5% of AA (20:4 ω6), 2% of LA (18:2 ω6), 21% of OA (18:1 ω9), and 12% of PA (16:0). To prevent lipid oxidation, the different oil-enriched diets were prepared weekly and kept in refrigerators (dark place and at 4°C). Dietary oils profile was analyzed by gas liquid chromatography (GLC) (13). The values obtained match those of the American Oil Chemists Society (14). After 3 months of feeding with dietary treatment, mice were euthanized with isoflurane to examine organs and tissues in suitable condition for multiple analyses. Another group of animals was allowed to die naturally to evaluate survival time (12). Animal studies were conducted in accordance with the guidelines of the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and all the procedures were approved by the Institutional Committee for the Care and Use of Laboratory Animals of the School of Medicine, University of Cordoba, Cordoba, Argentina (Project Number: V-21/2022).

Tumor volume, histopathology, fibrosis, metastasis, and apoptosis evaluation

The tumor pancreatic volume was measured with a digital caliper and calculated from the transverse (width) and longitudinal (length) diameters in millimeters of the tumor using the formula: Volume = Length × Width2/2 (12). All surgical organs (pancreas, liver, and lungs) were fixed with 10% formalin at 24°C for 24 hours. Tissues were embedded in paraffin and sliced to a thickness of 4 μm for H&E and Masson’s trichrome stains as well as immunohistochemistry as described below.

H&E staining and Masson’s trichrome staining

Histological analysis of tumor tissue sections involved two distinct staining protocols. H&E staining consisted of a 20-minute immersion in Mayer’s hematoxylin followed by 5 minutes in eosin. The Masson’s trichrome protocol began with tissue deparaffinization and rehydration through sequential alcohol gradients (100%, 95%, and 70%). After distilled water washing, tissues were processed through sequential steps: 10-minute staining in Weigert’s iron hematoxylin working solution, distilled water rinse, 15-minute staining in 1% Biebrich scarlet-acid fuchsin solution, differentiation in 5% phosphomolybdic-phosphotungstic acid solution, immediate 10-minute staining in aniline blue solution, 5-minute differentiation in 1% acetic acid solution, final distilled water rinse, rapid dehydration in 95% ethanol, and xylene clearing. Microscopic evaluation was performed using a Leica ICC50 HD microscope at 100× and 400× of magnification, examining 10 high-power fields per tissue sample (15).

Histopathological and metastases analysis

The pancreatic histopathological characteristics of control and Hω-3 groups were investigated using H&E and Masson’s trichrome stains. Architectural pancreatic tumor tissue was determined by levels of necrosis, fibrosis, vascularization, dysplastic glandular, and solid patterns. Tissue cytology was analyzed by anisokaryosis, anisocytosis, hyperchromatic, and prominent nucleoli. The prognostic determinants used were vascular permeation and perineural infiltration. All these parameters determined the cancer differentiation in the tumor tissues and graded them according to Lee et al. and Goto et al. (11,15). The histopathological analysis was performed by a double blind study on slides from 6 to 9 animals of each dietary condition. Number and sites of macro and micro metastasis were determined by a magnifying lens and optical microscopy, respectively (12,16). H&E stained liver and lung tissues from control and Hω-3 KPC mice were analyzed to determine the presence and number of metastases. Each tissue was evaluated in 10 high-power fields using a light microscope (Leica ICC50 HD microscope) (15).

Apoptosis and mitosis determination

Apoptosis in pancreatic tissue was determined by the terminal deoxyribose nucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method for the specific detection and quantification of apoptotic cells within a cell population using the Dead EndTM Colorimetric TUNEL System (Promega) in accordance with the manufacturer’s protocol. To detect mitotic figures, we conducted the Ki-67 staining since Ki-67 is a protein preferentially expressed during late G1-, S-, M-, and G2-phases of the cell cycle, while cells in the G0 (quiescent) phase are negative for this protein. The anti-Ki-67 (Dako Denmark A/S, DK-2600 Glostrup, Denmark) monoclonal antibody was used according to manufacturer protocol. Cells giving a nuclear brown positive precipitate were considered positive for TUNEL and Ki-67. Positive cells were counted in each tissue sample slide in a blinded manner from three animals for each of the dietary conditions per 10 high-power fields using a light microscope (Leica DM500, Toronto, Canada, 40× magnification). Then, they were analyzed with ImageJ 1.53c version for Windows (17). Results were expressed as a mean ± SEM of TUNEL or Ki-67 positive cells in an area of 5 × 10-3 mm2 (18).

Immunohistochemical detection of PPARγ and Shh

Paraffin-fixed Pancreatic tissue samples sections were prepared for Peroxisome proliferator-activated receptor gamma (PPARγ) and Shh expression by immunohistochemistry. We used PPARγ antibody (Abcam, Cambridge, UK, #sc-7196 1:100) and Human/Mouse Shh N‑Terminus Antibody (BioTechne, Systems, Minneapolis, MN, USA, #AF464 1:100). Horseradish peroxidase conjugated anti-rabbit was used as secondary antibody. Each tissue sample slide was evaluated in 10 high-power fields using a light microscope (100× and 400× magnification using Leica ICC50 HD microscopy). The positive cells (cells giving a nuclear brown positive precipitate) were quantified using digital micrographs obtained with an optical microscope (Leica ICC50 HD) and then analyzed with ImageJ 1.53c version for windows (17). Images were obtained from 10 randomly selected nonoverlapping fields of each section of tumor tissue at 100× and 400× magnification. Results were expressed as a mean ± SEM of PPARγ or Shh positive cells in an area of 5 × 10-3 mm2 (19). All experiments were performed in triplicate and repeated at least three times, unless otherwise stated.

Tumor cells suspensions

TC suspensions were obtained from primary tumors in necrosis free areas, blood clots, or connective tissue. Tissues were treated with 0.01% pronase and 0.24% Type1-deoxyribonuclease in Minimum Essential Media (Sigma-Aldrich, St. Louis, MO, USA), washed twice, resuspended in Ca2+ and Mg2+ free phosphate buffer saline (PBS), and stimulated with calcium ionophore A23187 (2 µM) (Sigma-Aldrich, St. Louis, MO, USA) for 15 min at 37ºC. An amount of 1 × 107 TC/ml was used for GLC and reverse phase high performance liquid chromatography (HPLC) (12). All experiments were performed in triplicate and repeated at least three times, unless otherwise stated.

Plasma membrane separation and fatty acid determination

The effect of dietary formulas on pancreatic cell membrane polyunsaturated fatty acids (PUFAs) profiles was evaluated using GLC analysis as previously described (20). Plasma membranes from tumor cell (TC) suspensions (2 × 106/ml) were obtained by homogenization in hypotonic Hepes-Manitol buffer using a Polytron homogenizer (7 seconds, setting 7), followed by 10-mM CaCl2 treatment and sequential centrifugation steps (3000 g for 15 minutes, supernatant at 48 000 g for 30 minutes). Plasma membrane fragments contained in the pellet were stored in PBS at 20°C overnight. Membrane lipids were extracted and partitioned following Folch methodology (21), obtaining methyl esters from the phospholipid fraction. Fatty acids methyl esters were analyzed employing a polyethylene glycol capillary column (30 m × 320 μm ID × 0.50 μm) in a Perkin Elmer Clarus 500 gas–liquid chromatograph with flame ionization detector (PerkinElmer Instruments, Waltham, MA, USA). Fatty acid identification relied on retention time comparison with authentic standards, while quantification used peak area calculations referenced to pure fatty acid standard curves (Nu-Chek Prep Inc, Elysian, MN, USA). Experimental procedures were performed in triplicate with minimum three repetitions, unless otherwise noted.

PUFAs metabolite analysis

The generation of 12(S)-Hydroxyeicosatetraenoic Acid [12(S)-HETE] from 12-Lipoxygenase with AA, ω-6, and 5-hydroperoxy-18-hydroxyeicosa-pentaenoic acid [5(S)-HEPE] with EPA ω-3, as substrate, were estimated as described in Kelavkar et al. (22). Determinations were made in 6–8 samples of TC suspensions (2 × 106 TC/ml) of each dietary treatment. Cells from primary tumor or culture cells treated with fatty acids, arachidonic acid (AA), EPA, or DHA 50 µM were suspended in PBS (Ca2+ and Mg2+) and stimulated with calcium ionophore A23187 to induce the release of eicosanoids. TC metabolites were extracted using a STRATA C-18 cartridge (1ml) (Phenomenex, Torrance, CA, USA) and detected by HPLC. The analysis was conducted with a C18 Phenosphere-Next column (5 µm, 4.6 × 250 mm) (Phenomenex) in a Beckman System Gold Programmable Module Model 126 (Beckman, Indianapolis, IN, USA). A linear gradient from solvent A: methanol: water: acetic acid, 50:50:0.02 (v/v/v), pH 6 to solvent B: methanol, for over 20 minutes. UV Beckman System Gold Model 166 with a programmable detector was also used. The analysis of the eicosanoids was obtained by using curves of the standards (Cayman Chemical, Ann Arbor, MI, USA) and was expressed as ng/1 × 107 TC (12,13,20). All experiments were performed in triplicate and repeated at least three times, unless otherwise stated.

Reverse transcription quantitative-PCR

Total RNA was extracted from tissue pancreatic tumor using TRIZOL reagent (Invitrogen, Waltham, MA, USA). Complementary DNA (cDNA) was generated from 5 μg of RNA by reverse transcription using M-MLV Reverse Transcriptase (Promega). Samples (2 μL) of cDNA from the synthesis reaction diluted 1:10 were used for PCR analysis. Samples were extracted from KPC mice fed with Control and Hω-3 diets. Expression analysis was performed using Real Mix B124-100 (Biodynamics, Ciudad Autonoma de Buenos Aires, Argentina), a 7500 Fast Real Time PCR Systems (Applied Biosystems, Waltham, MA, USA). Predesigned primers/probes for the expression of Shh: (forward) 5ʹ-gaagatcacaagaaactccgaacg-3ʹ, reverse: 5ʹ-tggattcatagtagacccagtcgaa-3ʹ) and Actinβ: (forward) 5´-gacggccaggtcatcactattg-3´, (reverse) 5´-aggaaggctggaaaagagc-3´ were purchased from Integrated DNA Technologies (Coralville, IA, USA). The amount of the transcript was calculated and expressed as the difference relative to the ACTB control gene (2ΔCt), where ΔCt represents the cycle threshold difference between the gene of interest and the control gene (23). All experiments were performed in triplicate and repeated at least three times, unless otherwise stated.

Statistical analysis

The data were described by mean values and SE (standard error). The comparison among groups was evaluated using analysis of variance (ANOVA) one-way, after corroborating the assumption of normality of the data, by Q-Q plot, and test by Shapiro Wilks (12). The Kaplan–Meier curve was analyzed using Gehan–Breslow–Wilcoxon test. A P-value < .05 was established for statistical significance. Data were analyzed using R Core Team (24) and InfoStat (25) software.

Results

Hω-3 dietary intervention reduced tumor volume and metastases and improved survival

In order to evaluate the Hω-3 intervention, KPC animals fed with cod oil-enriched diets as described in Fig. 1A. Macroscopy analysis showed that pancreatic tumor volume from KPC animals fed with Hω-3 enriched diets was significantly smaller than the tumors from control animals (306.42 ± 109.79 mm3 versus 2591.62 ± 668.49 mm3) (Fig. 1B and C) (P < .05). Animals fed with Hω-3 diet also showed a significant reduction in the metastases in lungs (3.00 ± 0.57 versus 6.25 ± 2.49) and in liver (0.10 ± 0.00 versus 6.20 ± 2.49) in comparison with the animals fed with control diet (Fig. 1D–G) (P < .02). Additionally, KPC mice fed with the Hω-3 enriched diet exhibited a trend toward improved survival rate (Fig. 1H) (P < .166).

Figure 1.

Experimental diagram and results showing dietary omega-3 fatty acids effects in KPC mice. The diagram comprises study design, images of pancreatic tumor specimens, tumor volume graphs, lung and liver metastases photomicrographs, metastasis quantification, and Kaplan Meier survival curve. Results demonstrate tumor suppression and reduced metastasis in treated group.

Open in a new tab

Dietary ω-3 fatty acids suppress pancreatic tumor growth and metastasis in the KPC mouse model. (A) Experimental design of in vivo model: since weaning (30 days after birth), KPC animals received two different diets sufficient in essential fatty acids: control group and the Hω-3 fatty acids content. The animals were sacrificed after 180 days of feeding with the experimental diets and the parameters of tumor pancreatic progression in mice were evaluated: tumor volume and metastases, biochemical and molecular tumor analysis, and histopathology. (B) Macroscopic representative view of pancreas from KPC mice fed with control and Hω-3 diets. (C) Bars represent pancreatic volume recorded after necropsy. The values were obtained from independent samples and represent the mean ± SEM (standard error of the mean); asterisks indicate significant differences compared with the control group (P < .05; n = 8–10). (D, E) Representative micrographs of metastases in lung and liver tissue observed in KPC mice fed with control and Hω-3 diets (H&E, 100× and 400×). (F, G) Bars represent the number of microscopic metastases in the lung and liver of KPC mice (mean ± SEM); asterisks indicate significant differences compared with the control group (P < .05; n = 6–8). (H) Kaplan–Meier curve survival (n = 8–12).

As control for efficacy of our dietary intervention, pancreatic tumor lipid membrane composition and eicosanoids release were evaluated with each dietary intervention in tissues of KPC mice (Fig. 2A and Supplementary Table 2) (P < .05). As expected, our analysis showed an increment in the percentage of EPA (1.27 ± 0.09 versus 0.00 ± 0.00) and DHA (1.94 ± 0.21 versus 1.36 ± 0.07) in TCs from KPC mice fed with Hω-3 PUFAs and a diminution of ω-6 PUFAs as AA (5.85 ± 0.26 versus 10.70 ± 2.74) respect to tumors from animals fed with control diet (Fig. 2A) (P < .05). The Hω-3 PUFAs induced an increment in 5(S)-HEPE (1.21 ± 0.13 versus 0.99 ± 0.01), an eicosanoid derived from EPA (Fig. 2B) (P < .05), and a decrease in 12(S)-HETE (0.186 ± 0.14 versus 0.302 ± 0.10), an eicosanoid derived from AA (ω-6) in KPC mice fed on Hω-3 PUFAs compared with control diet (Fig. 2C) (P < .05). Finally, we examined the expression of PPARγ, a molecule known to be induced by ω-3 fatty acids (especially EPA and its derivatives) (26). As shown in Fig. 2D and E Hω-3 dietary PUFAs significantly increased the tumor expression of PPARγ in Hω-3 fed KPC mice compared with the Control group fed diet (5.26 ± 1.01 versus 2.23 ± 0.86) (P < .05).

Figure 2.

Data showing membrane lipid alterations and molecular changes in KPC mice. Includes graphs with the quantitation of membrane fatty acids (EPA, DHA, AA), and eicosanoid measurements (5(S)-HEPE, 12(S)-HETE). PPARγ immunohistochemistry micrographs, and PPARγ-positive cell quantification. Results indicate significant membrane composition changes and increased PPARγ expression in omega-3 treated group.

Open in a new tab

Dietary ω-3 fatty acids modulate TC membrane lipid profile, eicosanoid metabolism, and PPARγ expression. (A) Bars represent the percentage of fatty acids in pancreatic TC membranes: the ω-3 EPA (C20:5) and DHA (C22:6) acids and the ω-6 AA (C20:4). Values represent mean ± SEM, asterisks indicate significant differences compared with the control group (P < .05; n = 3). (B, C) Bars represent the content (in ng) of eicosanoids in tumor membranes: 5(S)-HEPE, derived from the eicosapentaenoic acid (C20:5, ω-3 fatty acid) and 12(S)-HETE, derived from the AA (C20:4, ω-6 fatty acid). Values represent mean ± SEM (n = 3). (D) Representative micrographs at 100x and 400x magnifications showing PPARγ immunostaining in pancreatic tumor samples from KPC mice fed with Control and Hω-3 diets. (E) Bars represent the number of PPARγ-positive cells by immunohistochemistry in micrographs of pancreas sections from each dietary group. The values were obtained from independent samples divided into 10 sections each and represent the mean ± SEM; asterisks indicate significant differences compared with the control group (P < .05; n = 5)

Pancreatic tumors from Hω-3 diet group showed reduced tumor grade and increased cell death

Histopathological analysis of pancreatic tissue from KPC mice fed with Hω-3-enriched diets presented architectural changes in tissue differentiation parameters including necrosis (Fig. 3A1), vascularization (Fig. 3A2), dysplastic gland pattern, and solid pattern (Fig. 3A3) than pancreatic tumors from mice with fed with Control diets (P < .05). Indeed, tissue pancreatic cytology analysis of tumors from KPC mice fed with Hω-3-enriched diets resulted in less significantly grade of anisokaryosis, anisocytosis, hyperchromatic, and prominent nucleoli (Fig. 3A4 and A5) (P < .05) than tumors from the control diet fed animals. Also, pancreatic tissue from KPC mice fed with Hω-3 showed less perineural cancer cell infiltration and presence of vascular permeation (Fig. 3A2, A3, and A5) (Supplementary Fig. 1B and C) (P < .05). Hω-3 dietary treatment in KPC mice showed pancreatic affected pancreatic tissue and cellular parameters resulting in higher significant differentiation cancer grade respect to tumors from KPC mice fed with control diet (33.3% versus 83.0%) (Fig. 3B and Supplementary Fig. 1A) (P < .05). TUNEL analysis of tissues from KPC mice fed with an Hω-3 diet showed significant higher levels of apoptotic cells (31.77 ± 6.98 versus 10.21 ± 3.30) (Fig. 4A and B) (P < .01) without affecting cell proliferation as measured by Ki-67 signal (6.86 ± 1.69 versus 9.58 ± 2.37) (Fig. 4C and D) (P < .36).

Figure 3.

Histopathological analysis of pancreatic tumor architecture from control and omega-3 treated KPC mice. The figure illustrates features of H&E micrographs showing tissue characteristics, including necrosis, vascularization, glandular patterns, and nuclear features. It also includes graphs  with quantification of tumor differentiation grades, showing decreased percentage of poorly differentiated adenocarcinoma in treated group.

Open in a new tab

Histopathological assessment of pancreatic tumor differentiation in KPC mice fed an Hω-3 diet. (A) Representative H&E micrographs of pancreatic tissues from KPC mice after control and Hω-3 dietary treatment (100× and 400×). We analyzed architectural variables of pancreatic tumor tissues: (1) necrosis, (2) vascularization, (3) dysplastic glandular pattern and solid pattern, (4) anisokaryosis and anisocytosis, and (5) hyperchromatic and prominent nucleoli. Malignant typical lesions of high undifferentiated adenocarcinoma were observed in KPC mice of the control diet; moderately differentiated pancreatic adenocarcinoma and glands were observed in KPC animals from Hω-3 experimental group (H&E stain, 100× and 400×). (B) Bars represent cancer differentiation grade of pancreatic tumor observed in KPC mice: well-differentiated, moderately differentiated, and poorly differentiated pancreatic adenocarcinoma. Asterisks indicate significant differences compared with the control group (P < .05; n = 6–8)

Figure 4.

Apoptosis and proliferation analysis in pancreatic tissue from control and Hω-3 diets KPC mice. The figurea includes TUNEL assay micrographs and apoptosis quantification and Ki67 immunostaining micrographs with proliferation rate quantification. Data demonstrates enhanced apoptosis in tumors from animals in the Hω-3 diet group, while proliferation remains unchanged.

Open in a new tab

The effect of ω-3 fatty acid-rich diet on apoptosis and proliferation. (A) Representative micrographs of apoptotic cells detected by TUNEL in pancreatic sections (indicated by positive TUNEL staining) from KPC mice fed with control and Hω-3 diets (100x and 400x). (B) Bars represent the percentage of apoptotic cells in pancreatic tumor sections from each experimental group; the values were obtained from 10 independent sections per animal and represent the mean ± SEM; asterisks indicate significant differences compared to the Control group (P < 0.05; n=5). (C) Representative micrographs of Ki67 cells in pancreatic tumor sections from KPC mice fed with control and Hω-3 diets (100x and 400x). (D) Bars represent the percentage of proliferative cells in micrographs of pancreas sections (Ki67-positive cells) for each dietary group of mice; the values were obtained from independent samples divided into 10 sections each and represent the mean ± SEM (ns; n=5)

Omega-3-enriched diets reduced primary tumors fibrosis and Shh expression

Further analysis of this phenotype examining the fibrotic levels as area percentage of the signal of Masson stain in each histological section of pancreatic tumor showed that Hω-3 diet decreased significantly the fibrosis index in KPC animals with respect to control diet (17.11 ± 3.21 versus 40.61 ± 1.26) (Fig. 5A and B) (P < .01). Immunohistochemical analysis of potential mediators of this phenotype showed a significantly decreased expression of the ligand Shh protein in KPC animals fed with the Hω-3 diet compared with the control diet (7.70 ± 2.13 versus 26.31 ± 3.45) (Fig. 5C and D) (P < .05). Similarly, Shh expression evaluated by quantitative PCR (qPCR) was decreased in pancreatic tumors from KPC of Hω-3 dietary treatment in comparison with control diet (1.23 ± 0.40 versus 3.81 ± 0.97) (Fig. 5E) (P < .05).

Figure 5.

Microscopy images and quantitative analysis comparing pancreatic tissue samples from KPC mice fed with control versus Hω-3 diets. Masson staining shows fibrosis in light blue at two different magnifications, with corresponding quantification of fibrotic areas. Immunohistochemistry reveals Shh protein expression in brown at two magnifications, accompanied by positive cell count data. Gene expression analysis shows Shh mRNA levels measured by qPCR. Results demonstrate that Hω-3 diet reduces both pancreatic tumor fibrosis and Shh expression.

Open in a new tab

The impact of an ω-3 fatty acid-rich diet on fibrosis and SHH expression. (A) Representative micrographs of levels of fibrosis (indicated by Masson-positive signal) in pancreatic samples from KPC mice fed with control and Hω-3 diets (Masson stain, 100x and 400x). (B) Bars represent the percentage of fibrotic-positive areas (Masson-positive staining) in pancreatic tissues of KPC mice fed with control and Hω-3 diets; the values were obtained from independent samples divided into 10 sections each and represent the mean ± SEM; asterisks indicate significant differences compared to the control group (P < 0.05; n=10). (C) Representative micrographs of Shh expression detected by immunohistochemistry (indicated by positive immunostaining) in pancreatic samples from both experimental groups (100x and 400x). (D) Bars represent the number of Shh-positive cells by immunohistochemistry in micrographs of pancreas sections from each dietary group. The values were obtained from 10 independent tumor sections each and represent the mean ± SEM; asterisks indicate significant differences compared to the control group (P < 0.05; n=5). (E) Bars represent expression of Shh mRNA in pancreatic tumor tissue of KPC mice fed with control and Hω-3 diets determined using qPCR and is shown as Fold Change (mean ± SEM); asterisks indicate significant differences compared to control group (P < 0.05; n=3)

Discussion

In recent years, considerable progress has been made in the management of pancreatic cancer, such as the improvement of surgical procedures as well as adjuvant and neoadjuvant strategies (27). However, despite these advancements, current approaches are insufficient to effectively control this disease to improve patients’ outcome; thus further research is needed to develop new and more effective treatments (28,29). Recently, there is a growing interest in exploring the potential of natural and dietary products (nutraceuticals) as chemopreventive and treatment options for pancreatic cancer (30). While some clinical studies have provided insights into the potential benefits of ω-3 PUFAs interventions in pancreatic cancer, direct evidence of their anti-tumor activities in disease relevant models remain limited (31,32). In this work, we have demonstrated that ω-3 dietary fatty acids treatment can inhibit disease progression in a genetically engineered murine model closely mimicking human pancreatic cancer molecular and biological features. These include similarities in preneoplastic lesion (PanIN) development and progression, metastasis patterns, and microenvironment features such as desmoplastic stroma and other histopathological features. The model also provides a genetic background expressing mutant KRAS and TP53 at endogenous levels (as reported in human tumors) differing to other studies that used transgenic model overexpressing mutant KRAS (33). Nevertheless, we acknowledge that translating results to the clinical setting requires the testing of additional genetic backgrounds (e.g. loss of CDK2A or SMAD4) and potential differences in long-term microenvironment evolution that were not represented in this genetic model including level of TC clonal heterogeneity and different immune profile seen in patients (34,35).

The 7:1 ω-3:ω-6 ratio used in our work allows an effective cellular incorporation and increment of EPA and DHA as evidenced in the lipid profiles of the plasma membranes from pancreatic tumors of KPC animals fed with fish oil-enriched diets (Fig. 2A and Supplementary Table 2). Also, the inhibitory mechanism involves a significantly reduction of ω-6-PUFAs in the pancreatic cancer cells, particularly AA, and the reduction of its derivate pro-tumorigenic eicosanoid 12(S)-HETE (Fig. 2A and C) and an increment of the EPA derived eicosanoid, 5(S)-HEPE (Fig. 2B). Similar ratio was previously used by Roebuck et al., who demonstrated anticarcinogenic effects in chemically induced rat pancreatic preneoplastic lesions (36). Other researchers, such as Ding et al., using a lower ratio (2.5:1) in a genetic pancreatic cancer model, showed a reduction in proliferation associated with the AKT pathway. However, the study was focus on the study of the development precancerous lesions rather than pancreatic tumors incidence, and they did not analyze the incorporation levels of dietary fatty acids into the plasma membranes of these precancerous lesions through lipid profiling (33). Although several animal studies have explored ω-3 enriched diets with varying ratios (3), their methodological differences prevent direct comparisons. Nevertheless, our findings reveal notable pancreatic carcinogenesis inhibition that is accompanied by the regulation of specific molecular pathways. Our dietary intervention strategy encompassed both chemopreventive and therapeutic aspects, setting it apart from previous research that primarily examined precancerous lesions or utilized different experimental models.

This anti-tumoral effect seen in our study was accompanied by significant decrease in tumor fibrosis and levels of the profibrotic Hedgehog ligand Shh (Fig. 2D and E). Our results also show increased PPARγ levels, a transcription factor with known role in lipid metabolism, cellular differentiation, and inflammatory response in epithelial cells (37). These results are aligned with the studies from Gaiser et al. (38) and Cheng et al. (39) demonstrating tumor volume reduction through PPARγ activation by DHA and EPA. The Hω-3 dietary effect on Shh expression could be in part due to the inhibition of the ω-6 fatty acid cascade of eicosanoids generation, as supported by studies showing that inhibition of AA metabolism suppressed pancreatic cancer cell proliferation through Hedgehog pathway deregulation (40), while Wang et al. demonstrated that Hω-3 diets induce PPARγ expression and inhibit pancreatic stellate cells activation, reducing tumor fibrosis (41). Furthermore, Feng et al. showed that PPARγ can inhibit Shh expression through NFκB pathway suppression, leading to reduced fibrosis and tumor progression (42). Collectively, these findings highlight a novel molecular mechanism where ω-3 fatty acids could modulate the interplay between PPARγ and Shh pathways. This interaction appears to be crucial in mediating the anti-tumoral Hω-3 effects observed in pancreatic cancer. The upregulation of PPARγ and subsequent downregulation of Shh through ω-3 supplementation represents a promising therapeutic approach, particularly given its impact on both tumor progression and fibrosis reduction.

Conclusions

Our findings demonstrate that high concentrations of dietary ω-3 PUFAs, specifically EPA and DHA, can effectively reduce pancreatic tumor volume and metastasis, histopathological malignant characteristics, and a trend toward improved survival rate. These results contribute to the understanding of the cellular and molecular basis of pancreatic carcinogenesis impacted by dietary PUFAs and their derivatives, and offer a new alternative to treat this malignancy to improve patients’ outcome.

Supplementary Material

bgae081_suppl_Supplementary_Figure_S1
bgae081_suppl_supplementary_figure_s1.jpeg (833KB, jpeg)
bgae081_suppl_Supplementary_Table_S1
bgae081_suppl_supplementary_table_s1.docx (15.5KB, docx)
bgae081_suppl_Supplementary_Table_S2
bgae081_suppl_supplementary_table_s2.docx (17.5KB, docx)

Acknowledgements

We are indebted to Verónica Leticia Díaz and Sonia Elizabeth Alza for their help with the animal care, to Gina Mazzudulli for her help with immunohistological studies, and to Sara Lejter Manzur for her assistance with figures and graphical designs.

Contributor Information

María I Garay, Instituto de Investigaciones en Ciencias de la Salud, INICSA (CONICET - FCM UNC), 5016 Córdoba, Argentina; Instituto de Biología Celular y Cátedra de Biología Celular, Histología y Embriología, FCM-UNC, 5016 Córdoba, Argentina.

Tamara Mazo, Instituto de Investigaciones en Ciencias de la Salud, INICSA (CONICET - FCM UNC), 5016 Córdoba, Argentina; Instituto de Biología Celular y Cátedra de Biología Celular, Histología y Embriología, FCM-UNC, 5016 Córdoba, Argentina.

Victoria Ferrero, Instituto de Investigaciones en Ciencias de la Salud, INICSA (CONICET - FCM UNC), 5016 Córdoba, Argentina; Instituto de Biología Celular y Cátedra de Biología Celular, Histología y Embriología, FCM-UNC, 5016 Córdoba, Argentina.

Nelso N Barotto, Instituto de Biología Celular y Cátedra de Biología Celular, Histología y Embriología, FCM-UNC, 5016 Córdoba, Argentina.

Clarisa Lagares, Instituto de Investigaciones en Ciencias de la Salud, INICSA (CONICET - FCM UNC), 5016 Córdoba, Argentina.

María F Granton, Instituto de Biología Celular y Cátedra de Biología Celular, Histología y Embriología, FCM-UNC, 5016 Córdoba, Argentina.

María J Moreira-Espinoza, Instituto de Biología Celular y Cátedra de Biología Celular, Histología y Embriología, FCM-UNC, 5016 Córdoba, Argentina.

David C Cremonezzi, Departamento de Patología, Hospital Nacional de Clínicas, FCM-UNC, 5000 Córdoba, Argentina.

Andrea Comba, Department of Pathology, Division Neuropathology, Heersink School of Medicine, University of Alabama at Birmingham, Birmingham, AL, 35294, United States.

Mabel N Brunotto, Instituto de Investigaciones en Ciencias de la Salud, INICSA (CONICET - FCM UNC), 5016 Córdoba, Argentina; Departamento de Biología Bucal, Facultad de Odontología, UNC, 5016 Córdoba, Argentina.

Ezequiel J Tolosa, Schulze Center for Novel Therapeutics, Mayo Clinic, Rochester, MN 55905, United States.

Martín E Fernandez-Zapico, Schulze Center for Novel Therapeutics, Mayo Clinic, Rochester, MN 55905, United States.

María E Pasqualini, Instituto de Investigaciones en Ciencias de la Salud, INICSA (CONICET - FCM UNC), 5016 Córdoba, Argentina; Instituto de Biología Celular y Cátedra de Biología Celular, Histología y Embriología, FCM-UNC, 5016 Córdoba, Argentina.

Author contributions

M.I.G. conceived the study; T.M. made graphs and organized results; V.F. data analysis and curation; N.N.B. provided project conceptualization and assisted with chromatography techniques; C.L. monitored animal care; M.F.G. assisted with the immunohistological techniques; M.J.M.-E. assisted with the qPCR technique; D.C.C. performed the pathological analysis; A.C. and M.N.B. interpreted the results and performed statistical data analysis; E.J.T. assisted with animal breeding and care; M.E.F.-Z. participated in the project conceptualization and data analysis, and wrote the manuscript; M.E.P. designed the experiments, procured funding, participated in the data analysis, and wrote manuscript. All authors have read and approved the final version of the manuscript.

Ethics Statement

The Institutional Committee for the Care and Use of Laboratory Animals of the School of Medicine, University of Cordoba, Argentina approved all the procedures (Number Project: V-21/2022).

Conflict of interest

None declared.

Funding

This work was supported by Agencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación (Grant FONCYT-PICT-2021-GRFTI-00543), Consejo Nacional de Investigaciones Científicas y Técnicas - Argentina (CONICET Grant #22920180100043CO), Secretaría de Ciencia y Tecnología de la Universidad Nacional de Córdoba (SECyT-UNC Grant 2018-2020: 411/18, 455/18 472/18; SECYT-UNC Grant 2023-258-E-UNC-SECYT#ACTIP), and Mayo Clinic Cancer Center.

Data Availability

The data underlying this article are available in the article and in its online supplementary material.

References

  • 1. Siegel  RL, Giaquinto  AN, Jemal  A.  Cancer statistics, 2024. CA Cancer J Clin  2024;74:12–49. https://doi.org/ 10.3322/caac.21820 [DOI] [PubMed] [Google Scholar]
  • 2. Suda  A, Umaru  BA, Yamamoto  Y  et al.  Polyunsaturated fatty acids-induced ferroptosis suppresses pancreatic cancer growth. Sci Rep  2024;14:4409. https://doi.org/ 10.1038/s41598-024-55050-4 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. Wirkus  J, Ead  AS, Mackenzie  GG.  Impact of dietary fat composition and quantity in pancreatic carcinogenesis: recent advances and controversies. Nutr Res  2021;88:1–18. https://doi.org/ 10.1016/j.nutres.2020.12.018 [DOI] [PubMed] [Google Scholar]
  • 4. Hingorani  SR, Wang  L, Multani  AS  et al.  Trp53R172H and KrasG12D cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice. Cancer Cell  2005;7:469–83. https://doi.org/ 10.1016/j.ccr.2005.04.023 [DOI] [PubMed] [Google Scholar]
  • 5. Dawson  DW, Hertzer  K, Moro  A  et al.  High-fat, high-calorie diet promotes early pancreatic neoplasia in the conditional KrasG12D mouse model. Cancer Prev Res (Phila)  2013;6:1064–73. https://doi.org/ 10.1158/1940-6207.CAPR-13-0065 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. Ead  AS, Wirkus  J, Matsukuma  K  et al.  A high-fat diet induces changes in mesenteric adipose tissue accelerating early-stage pancreatic carcinogenesis in mice. J Nutr Biochem  2024;131:109690. https://doi.org/ 10.1016/j.jnutbio.2024.109690 [DOI] [PubMed] [Google Scholar]
  • 7. Montecillo-Aguado  M, Tirado-Rodriguez  B, Huerta-Yepez  S.  The involvement of polyunsaturated fatty acids in apoptosis mechanisms and their implications in cancer. Int J Mol Sci  2023;24:11691. https://doi.org/ 10.3390/ijms241411691 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. Stephenson  JA, Al-Taan  O, Arshad  A  et al.  The multifaceted effects of omega-3 polyunsaturated fatty acids on the hallmarks of cancer. JLipids  2013;2013:1–13. https://doi.org/ 10.1155/2013/261247 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Fukui  M, Kang  KS, Okada  K  et al.  EPA, an omega-3 fatty acid, induces apoptosis in human pancreatic cancer cells: role of ROS accumulation, caspase-8 activation, and autophagy induction. J Cell Biochem  2013;114:192–203. https://doi.org/ 10.1002/jcb.24354 [DOI] [PubMed] [Google Scholar]
  • 10. Zou  HY, Zhang  HJ, Zhao  YC  et al.  N-3 PUFA deficiency aggravates streptozotocin-induced pancreatic injury in mice but dietary supplementation with DHA/EPA protects the pancreas via suppressing inflammation, oxidative stress and apoptosis. Mar Drugs  2023;21:39. https://doi.org/ 10.3390/md21010039 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. Lee  JW, Komar  CA, Bengsch  F  et al.  Genetically engineered mouse models of pancreatic cancer: the KPC model (LSL‐Kras G12D/+;LSL‐Trp53R172H/+;Pdx‐1‐Cre), its variants, and their application in immuno‐oncology drug discovery. Curr Protoc Pharmacol  2016;73:14.39.1–14.39.20. https://doi.org/ 10.1002/cpph.2 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. Garay  MI, Comba  A, Vara Messler  M  et al.  Tumorigenic effect mediated by ROS/eicosanoids and their regulation on TP53 expression in a murine mammary gland adenocarcinoma. Prostaglandins Other Lipid Mediat  2021;155:106564. https://doi.org/ 10.1016/j.prostaglandins.2021.106564 [DOI] [PubMed] [Google Scholar]
  • 13. Comba  A, Almada  LL, Tolosa  E  et al.  Nuclear factor of activated T cells-dependent downregulation of the transcription factor glioma-associated protein 1 (GLI1) underlies the growth inhibitory properties of arachidonic acid. J Biol Chem  2016;291:1933–47. https://doi.org/ 10.1074/jbc.M115.691972 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. American Oil Chemists’ Society. Saturated, cis-monounsaturated, and cis-polyunsaturated fatty acids in marine and other oils containing long chain polyunsaturated fatty acids (PUFAs) by capillary GLC. In: Official Methods and Recommended Practices of the American Oil Chemists’ Society. 5th edn. Champaign (IL): AOCS Press, 1998 [Google Scholar]
  • 15. Goto  S, Yoshizawa  T, Ishido  K  et al.  Use of time‑density curves of dynamic contrast‑enhanced computed tomography for determination of the histological therapeutic effects of neoadjuvant chemotherapy for pancreatic ductal adenocarcinoma. Oncol Rep  2023;49:61. https://doi.org/ 10.3892/or.2023.8498 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. García  CP, Lamarque  AL, Comba  A  et al.  Synergistic anti-tumor effects of melatonin and PUFAs from walnuts in a murine mammary adenocarcinoma model. Nutrition  2015;31:570–7. https://doi.org/ 10.1016/j.nut.2014.06.001 [DOI] [PubMed] [Google Scholar]
  • 17. Rasband  WS, Schindelin  J, Arganda-Carreras  I  et al.  ImageJ. Bethesda: National Institutes of Health, 1997-2018. https://imagej.net/ij/ [Google Scholar]
  • 18. Kyrylkova  K, Kyryachenko  S, Leid  M  et al.  Detection of apoptosis by TUNEL assay. In: Kioussi  C (ed.), Odontogenesis. Methods in Molecular Biology. Totowa, NJ: Humana Press, 2012, 41–7. https://doi.org/ 10.1007/978-1-61779-860-3_5 [DOI] [PubMed] [Google Scholar]
  • 19. Scherma  ME, Madzzuduli  G, Silva  RA  et al.  The effects of ω−6 and ω−3 fatty-acids on early stages of mice DMBA submandibular glands tumorigenesis. Prostaglandins Leukot Essent Fatty Acids  2017;125:48–55. https://doi.org/ 10.1016/j.plefa.2017.08.004 [DOI] [PubMed] [Google Scholar]
  • 20. Vara-Messler  M, Pasqualini  ME, Comba  A  et al.  Increased dietary levels of α-linoleic acid inhibit mammary tumor growth and metastasis. Eur J Nutr  2017;56:509–19. https://doi.org/ 10.1007/s00394-015-1096-6 [DOI] [PubMed] [Google Scholar]
  • 21. Folch  J, Lees  M, Sloane Stanley  GH.  A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem  1957;226:497–509. https://doi.org/ 10.1016/S0021-9258(18)64849-5 [DOI] [PubMed] [Google Scholar]
  • 22. Kelavkar  UP, Hutzley  J, McHugh  K  et al.  Prostate tumor growth can be modulated by dietarily targeting the 15-lipoxygenase-1 and cyclooxygenase-2 enzymes. Neoplasia  2009;11:692–9. https://doi.org/ 10.1593/neo.09334 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. Lo Ré  AE, Fernández-Barrera  MG, Almada  LL  et al.  Novel AKT1-GLI3-VMP1 pathway mediates KRAS oncogene-induced autophagy in cancer cells. J Biol Chem  2012;287:25325–34. https://doi.org/ 10.1074/jbc.M112.370809 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. R Core Team. R: a language and environment for statistical computing . In: R Foundation for Statistical Computing. Vienna: R Foundation, 2024 [Google Scholar]
  • 25. Di Rienzo  JA, Casanoves  F, Balzarini  MG  et al.  InfoStat versión 2012. Córdoba: Grupo InfoStat, FCA, Universidad Nacional de Córdoba, 2012. [Google Scholar]
  • 26. Yang  X, Li  X, Hu  M  et al.  EPA and DHA differentially improve insulin resistance by reducing adipose tissue inflammation—targeting GPR120/PPARγ pathway. J Nutr Biochem  2024;130:109648. https://doi.org/ 10.1016/j.jnutbio.2024.109648 [DOI] [PubMed] [Google Scholar]
  • 27. Mizrahi  JD, Surana  R, Valle  JW  et al.  Pancreatic cancer. Lancet  2020;395:2008–20. https://doi.org/ 10.1016/S0140-6736(20)30974-0 [DOI] [PubMed] [Google Scholar]
  • 28. Barrios  P, Chawla  A.  Gastrointestinal malignancies: pancreatic cancer clinical trials in neoadjuvant chemotherapy. In: Bentrem  D, Benson  AB (eds.) Gastrointestinal Malignancies. Cancer Treatment and Research. Cham: Springer Nature Switzerland, 2024, 119–29. https://doi.org/ 10.1007/978-3-031-61238-1_6 [DOI] [PubMed] [Google Scholar]
  • 29. Vitello  D, Talamonti  MS.  Modern treatment strategies for borderline resectable pancreatic cancer. In: Bentrem  D, Benson  AB (eds.), Gastrointestinal Malignancies. Cancer Treatment and Research. Cham: Springer Nature Switzerland, 2024, 67–88. https://doi.org/ 10.1007/978-3-031-61238-1_4 [DOI] [PubMed] [Google Scholar]
  • 30. Kim  A, Ha  J, Kim  J  et al.  Natural products for pancreatic cancer treatment: from traditional medicine to modern drug discovery. Nutrients  2021;13:3801. https://doi.org/ 10.3390/nu13113801 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31. Abe  K, Uwagawa  T, Haruki  K  et al.  Effects of ω-3 fatty acid supplementation in patients with bile duct or pancreatic cancer undergoing chemotherapy. Anticancer Res  2018;38:2369–75. https://doi.org/ 10.21873/anticanres.12485 [DOI] [PubMed] [Google Scholar]
  • 32. Isherwood  J, Arshad  A, Chung  WY  et al.  Myeloid derived suppressor cells are reduced and T regulatory cells stabilised in patients with advanced pancreatic cancer treated with gemcitabine and intravenous omega 3. Ann Transl Med  2020;8:172–2. https://doi.org/ 10.21037/atm.2020.02.02 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. Ding  Y, Mullapudi  B, Torres  C  et al.  Omega-3 fatty acids prevent early pancreatic carcinogenesis via repression of the AKT pathway. Nutrients  2018;10:1289. https://doi.org/ 10.3390/nu10091289 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. Ying  H, Kimmelman  AC, Bardeesy  N  et al.  Genetics and biology of pancreatic ductal adenocarcinoma. Genes Dev  2024;38:11–48. https://doi.org/ 10.1101/gad.351863.124 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35. Vudatha  V, Herremans  KM, Freudenberger  DC  et al.  In vivo models of pancreatic ductal adenocarcinoma. In: Emdad  L, Atfi  A, Gogna  R  et al. (eds.), Advances in Cancer Research. Amsterdam: Elsevier, 2023, 75–112. https://doi.org/ 10.1016/bs.acr.2023.02.002 [DOI] [PubMed] [Google Scholar]
  • 36. O’Connor  TP, Roebuck  BD, Peterson  FJ  et al.  Effect of dietary omega-3 and omega-6 fatty acids on development of azaserine-induced preneoplastic lesions in rat pancreas. J Natl Cancer Inst  1989;81:858–63. https://doi.org/ 10.1093/jnci/81.11.858 [DOI] [PubMed] [Google Scholar]
  • 37. Das  P, Dutta  A, Panchali  T  et al.  Advances in therapeutic applications of fish oil: a review. MeasFood  2024;13:100142. https://doi.org/ 10.1016/j.meafoo.2024.100142 [DOI] [Google Scholar]
  • 38. Gaiser  RA, Pessia  A, Ateeb  Z  et al.  Integrated targeted metabolomic and lipidomic analysis: a novel approach to classifying early cystic precursors to invasive pancreatic cancer. Sci Rep  2019;9:10208. https://doi.org/ 10.1038/s41598-019-46634-6 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39. Cheng  S, Wang  G, Wang  Y  et al.  Fatty acid oxidation inhibitor etomoxir suppresses tumor progression and induces cell cycle arrest via PPARγ-mediated pathway in bladder cancer. Clin Sci  2019;133:1745–58. https://doi.org/ 10.1042/CS20190587 [DOI] [PubMed] [Google Scholar]
  • 40. Song  L, Chen  X, Wang  P  et al.  Effects of baicalein on pancreatic cancer stem cells via modulation of sonic Hedgehog pathway. Acta Biochim Biophys Sin  2018;50:586–96. https://doi.org/ 10.1093/abbs/gmy045 [DOI] [PubMed] [Google Scholar]
  • 41. Wang  Z, Dong  S, Zhou  W.  Pancreatic stellate cells: key players in pancreatic health and diseases (Review). Mol Med Rep  2024;30:109. https://doi.org/ 10.3892/mmr.2024.13233 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42. Feng  P, Xu  Y, Tong  B  et al.  Saikosaponin a attenuates hyperlipidemic pancreatitis in rats via the PPAR‑γ/NF‑κB signaling pathway. Exp Ther Med  2020;19:1203–12. https://doi.org/ 10.3892/etm.2019.8324 [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

bgae081_suppl_Supplementary_Figure_S1
bgae081_suppl_supplementary_figure_s1.jpeg (833KB, jpeg)
bgae081_suppl_Supplementary_Table_S1
bgae081_suppl_supplementary_table_s1.docx (15.5KB, docx)
bgae081_suppl_Supplementary_Table_S2
bgae081_suppl_supplementary_table_s2.docx (17.5KB, docx)

Data Availability Statement

The data underlying this article are available in the article and in its online supplementary material.

Articles from Carcinogenesis are provided here courtesy of Oxford University Press

RESOURCES