Abstract
Introduction:
Interleukin (IL)-28, a type III interferon related to the IL-10 family, shares multiple structural and functional characteristics with IL-22, another member of the IL-10 cytokine family. Previous research identified IL-22 as a downstream effector of IL-23 in colorectal tumorigenesis. However, data on IL-28 expression in colorectal cancer (CRC) and its relation to clinicopathological features, prognosis, and IL-23 expression remain limited.
Methods:
IL-23 and IL-28 immunohistochemical reactivity was evaluated in 75 specimens of colorectal mucinous adenocarcinoma (MA) and 75 specimens of non-mucinous adenocarcinoma (NMA) using the high-density manual tissue microarray method. Clinicopathological features and survival data were statistically analyzed.
Results:
MA exhibited higher IL-28 and IL-23 expression than NMA; however, this was statistically significant only for IL-23. IL-23 and IL-28 positivity rates showed a highly significant interrelation in MA only. Within the NMA group, no significant association was observed between IL-23 or IL-28 expression and clinicopathological features or survival. In the MA group, high IL-23 expression was significantly associated with positive lymphovascular invasion, advanced pathological tumor (pT) stage, late TNM stage, and decreased disease-free survival and overall survival. High IL-28 expression was significantly associated with advanced pT stage, lymph node spread, advanced TNM stage, and decreased disease-free survival and overall survival.
Conclusion:
For the first time, the expression of IL-23 and IL-28 has shown a highly significant interrelation in MA patients, suggesting a possible interplay between them. High IL-23 and IL-28 expressions may have adverse prognostic effects on survival in MA. Molecular studies are necessary to further investigate the interaction between IL-23 and IL-28 in CRC.
Key Words: IL-23- IL-28, Mucinous, Non-mucinous, Colorectal adenocarcinoma
Introduction
Colorectal adenocarcinoma is the third most common malignancy worldwide, with limited treatment options [1, 2]. Mucinous adenocarcinoma (MA) is a distinct subtype of colorectal adenocarcinoma in which at least half of the neoplasm consists of extracellular mucinous pools (colloid carcinoma) or signet ring cells with intracellular mucin (signet ring carcinoma). It accounts for 5-20% of CRC cases [3]. MA differs from non-mucinous adenocarcinoma (NMA) in clinical, morphological, and molecular features and has been long associated with a poor therapeutic response and an unfavorable prognosis [4-6]. Previous research has demonstrated that inflammatory mediators regulate multiple critical aspects of carcinogenesis, including tumor cell proliferation, survival, and dissemination [2].
IL-23, a member of the interleukin-12 superfamily [7], consists of a unique p19 subunit and a common p40 subunit, which is shared with its relative IL-12. It is primarily secreted by activated dendritic cells and macrophages, playing a role in mucosal immunity [8, 9]. IL-23 has been implicated in the progression of chronic inflammation through the activation of T helper 17 (Th17) cells [10] and has been associated with several autoimmune disorders, including inflammatory bowel disease (IBD) [8], multiple sclerosis [11], and rheumatoid arthritis [12]. Recent studies suggest that IL-23-mediated effects contribute to tumor dissemination in various tissues. For example, IL-23 promoted liver cancer cell dissemination through the upregulation of matrix metalloproteinase 9 (MMP-9) [13]. Moreover, an anti-IL-23p19 monoclonal antibody significantly inhibited the spread of lung carcinoma cells in a mouse model, indicating a pro-metastatic role of IL-23 [14].
IL-28, known as IFN-λ, belongs to the recently identified type III IFN family [15, 16]. It is primarily secreted by epithelial cells following viral infection or by plasmacytoid dendritic cells. Other sources include antigen-presenting cells, Th17 cells, and neutrophils [17, 18]. IFN-λ has been suggested to exert dual effects in malignancy, with its anti-cancer properties demonstrated in specific tumor models, such as melanoma, lung cancer, and hepatocellular carcinoma [19, 20]. Conversely, IFN-λ has been shown to promote the dissemination of bladder carcinoma cells by upregulating matrix metalloproteinase 9 (MMP-9) [21]. Furthermore, IFN-λ induces angiogenesis, epithelial-mesenchymal transition, and cancer metastasis through a STAT3-dependent mechanism [22]. IFN-λ signals through its receptor, composed of IFNLR1 and IL-10R2. IL-10R2 is a receptor subunit shared by members of the IL-10 family [23, 24]. Therefore, IFN-λ is considered part of the IL-10 superfamily [25-27].
In CRC studies, IL-22 has been identified as a downstream effector of IL-23 in the development of colon carcinogenesis [28-31]. IL-22, a member of the IL-10 cytokine superfamily, signals through the IL-10R2 chain, which is overexpressed in CRC tissues and utilized by tumor cells to activate STAT3, promoting survival and proliferation. Among the ligands for IL-10R2, in addition to IL-10 and IL-22, IFN-λ shares this receptor chain [32]. Data regarding the role of IFN-λ in CRC are limited and warrant further investigation [2]. Moreover, the relationship between IL-23 and IL-28 in CRC has not yet been described. The present study aimed to investigate the relationship between IL-23 and IL-28 in CRC and their association with the clinicopathological features and prognosis of colorectal adenocarcinoma by evaluating their immunohistochemical expression in colorectal MA and NMA using the manual tissue microarray (TMA) method.
Materials and Methods
Cases
This retrospective study was conducted on 150 resected CRC specimens obtained from the Gastroenterology Center in Mansoura, Egypt, between 2007 and 2011. These included 75 specimens of MA (56 mucoid adenocarcinomas and 19 signet ring cell carcinomas) and 75 specimens of NMA (47 ordinary adenocarcinomas and 28 adenocarcinomas with mucoid differentiation < 50%). Clinico-pathological data were reviewed from patients’ files and included age, sex, tumor site, size, gross appearance, histological type, microscopic tumor borders, lymphovascular emboli, perineural invasion, pathological T stage, lymph node (LN) spread, distant metastasis (M), and TNM stage. All slides were re-evaluated. During case selection, cases with insufficient clinicopathological information, pre-operative therapy, or those composed entirely of hypocellular mucin lakes were excluded.
Tissue microarray design
Three TMA paraffin blocks were constructed using the mechanical pencil tip method and its modification, as previously described by Shebl et al. [33] and Foda [34]. For each specimen, three cores, each 0.8 mm in diameter, were obtained. Fifty cores of non-neoplastic colorectal mucosa were included as controls. Sections of 4 μm thickness were prepared for hematoxylin and eosin staining and immunostaining.
Immunostaining
Sections were deparaffinized and then incubated with 0.3% hydrogen peroxide in methanol for 30 minutes. This was followed by heating in an EDTA buffer solution (pH = 8.0) for 30 minutes using a microwave. Subsequently, the indirect immunoperoxidase method was performed using anti-IL-23 antibody (ab45420, rabbit polyclonal, Abcam; 1:100 dilution) and anti-IL-28 antibody (BZ16031, rabbit polyclonal, Bioworld Technology, Inc.; 1:100 dilution). Primary antibodies were applied for 30 minutes at room temperature. The immunoperoxidase technique was then carried out using the ImmunoPure Ultra-Sensitive ABC Peroxidase (catalog no. 32052; Thermo Scientific, UK), with diaminobenzidine as the chromogen.
Assessment of Immunostaining
The immunoreactivity of IL-23 and IL-28 was semi-quantitatively evaluated for each specimen. For both IL-23 and IL-28, the intensity and extent of positive tumor cells were scored for each core. Cytoplasmic and/or nuclear immunoreactivity was considered positive. The authors independently assessed immunostaining. Immunoreactivity intensity was scored as follows: 0 = no staining, 1 = faint staining, 2 = moderate, and 3 = strong. The proportion of positive neoplastic cells was graded as follows: 0 = 0% positive tumor cells, 1 = 1-10%, 2 = 11-50%, 3 = 51-80%, and 4 = > 80%. The intensity score was multiplied by the proportion score to calculate a final immunohistochemical score (IHS) ranging from 0 to 12. IHS scores were interpreted as follows: 9-12 = strong immunoreactivity, 5-8 = moderate, 1-4 = weak, and 0 = negative [35]. The mean score for each core of each specimen was calculated. For data interpretation, cases were classified into a low expression group (score = 0-4) and a high expression group (score = 5-12).
Statistical methodology
Data were analyzed using the SPSS statistical package version 25 (SPSS Inc., Chicago, IL). Numerical data were expressed as mean and standard deviation or median and range, as appropriate. Qualitative data were expressed as frequency and percentage. The chi-square test (Fisher’s exact test) was used to assess significant differences between MA and NMA cases in clinicopathological and histological features, as well as the relationship of IL-23 and IL-28 expression with these features within each group. The Kaplan-Meier method was used to estimate survival probabilities, and the log-rank test was applied to compare survival probabilities between the two groups. All tests were two-tailed, and a p-value ≤ 0.05 was considered significant. Last seen or death was considered the endpoint for cases. Disease-free survival (DFS) was defined as the period between diagnosis and the occurrence of disease recurrence (or the date last seen disease-free). Overall survival (OS) was defined as the period between diagnosis and the date last seen or death.
Results
The current study included 150 specimens of colonic adenocarcinoma. Ages ranged from 20 to 80 years (mean = 52.7 years). The study comprised 93 males and 57 females. The clinicopathological and histopathological features of 75 MA specimens and 75 NMA specimens are presented in Table 1. MA was significantly associated with older age (P = 0.017), advanced pT stage (P = 0.008), advanced pN stage (P = 0.008), and decreased tumoral neutrophils (P < 0.001) compared to NMA. All cases were stained for IL-23 and IL-28. Immunohistochemical expression of IL-23 and IL-28 was detected as cytoplasmic and/or nuclear brown staining in tumor cells (Figures 1 and 2). MA exhibited higher expression of IL-23 (68%) and IL-28 (65.3%) compared to NMA (50.7% and 52%, respectively); however, this difference was statistically significant only for IL-23 (P = 0.030) (Table 2). A highly significant association between IL-23 and IL-28 positivity rates was observed in MA cases (P < 0.001), but not in NMA cases (P = 0.16). Approximately 82.4% of MA cases with high IL-23 expression showed high IL-28 expression (Table 3). The relationship between IL-23 and IL-28 expression and clinicopathological and histological features was examined within each group. In the NMA group, no significant association was found between IL-23 or IL-28 expression and any clinicopathological characteristics. In contrast, in the MA group, high IL-23 expression was significantly associated with positive lymphovascular emboli (P = 0.05), advanced pT stage (P = 0.01), and advanced TNM stage (P = 0.03). High IL-28 expression was significantly associated with advanced pT stage (P = 0.001), nodal spread (P = 0.008), and advanced TNM stage (P = 0.02) (Tables 3 and 4). The remaining parameters showed no significant relationship with IL-23 or IL-28 expression within either group (data not shown).
Table 1.
Clinicopathological Features of 150 Cases of Mucinous & Non-Mucinous Colorectal Carcinomas
| Non- mucinous (a) | mucinous (b) | chi-square | P Value | |||
|---|---|---|---|---|---|---|
| No. | (%) | No. | (%) | |||
| Age | ||||||
| <40 | 10 | 13.3% | 22 | 29.3% | 5.720 | 0.017* |
| ≥40 | 65 | 86.7% | 53 | 70.7% | ||
| Gender | ||||||
| Male | 48 | 64.0% | 45 | 60.0% | 0.255 | 0.614 |
| Female | 27 | 36.0% | 30 | 40.0% | ||
| Tumor size | ||||||
| <6 | 46 | 61.3% | 36 | 48.0% | 2.690 | 0.101 |
| ≥6 | 29 | 38.7% | 39 | 52.0% | ||
| Depth of invasion | ||||||
| T1/T2 | 18 | 24% | 6 | 8% | 7.143 | 0.008* |
| T3/T4 | 57 | 76% | 69 | 92% | ||
| LN | ||||||
| N0 | 38 | 50.7% | 27 | 36.0% | 9.729 | 0.008* |
| N1 | 25 | 33.3% | 19 | 25.3% | ||
| N2 | 12 | 16.0% | 29 | 38.7% | ||
| TNMstage | ||||||
| I/II | 37 | 49.3% | 27 | 36% | 2.725 | 0.099 |
| III/IV | 38 | 50.7% | 48 | 64% | ||
| Location | ||||||
| Right | 18 | 24% | 23 | 30.7% | 0.887 | 0.829 |
| Left | 6 | 8% | 6 | 8% | ||
| Recto-sigmoid | 45 | 60% | 41 | 54.7% | ||
| Transverse | 6 | 8% | 5 | 6.7% | ||
| Gross picture | ||||||
| Fungating | 34 | 45.3% | 27 | 36% | 2.218 | 0.33 |
| Ulcerating | 19 | 25.3% | 27 | 36% | ||
| Annular | 22 | 29.3% | 21 | 28% | ||
| Lymphovascular emboli | ||||||
| Negative | 30 | 40% | 23 | 30.7% | 1.430 | 0.232 |
| Positive | 45 | 60% | 52 | 69.3% | ||
| Perineural invasion | ||||||
| Negative | 54 | 72% | 48 | 64% | 1.103 | 0.294 |
| Positive | 21 | 28% | 27 | 36% | ||
| Tumor margins | ||||||
| Budding | 68 | 90.7% | 69 | 92% | 0.084 | 0.772 |
| Pushing | 7 | 9.3% | 6 | 8% | ||
LN, lymph node; TNM, tumor-node-metastases; *P value ≤0.05 is significant; **P <0.001 is highly significant; a, Non-mucinous group includes 47 cases of ordinary adenocarcinoma & 28 cases of adenocarcinoma with mucinous component <50 %; b, Mucinous group includes 56 cases of mucoid adenocarcinoma & 19 cases of signet ring carcinoma
Figure 1.
IL-23 Expression in CRC (×200). (A) Weak staining in NMA. (B) Moderate staining in NMA (C) Strong staining in NMA. (D) Weak staining in MA. (E) Moderate staining in MA (F) Strong staining in MA.
Figure 2.
IL-28 Expression in CRC (×200). (A) Weak staining in NMA. (B) Moderate staining in NMA (C) Strong staining in NMA. (D) Weak staining in MA. (E) Moderate staining in MA (F) Strong staining in MA.
Table 2.
IL-23 & IL-28 Expressions in NMA & MA
| NMA No. (%) | MA No. (%) | Chi-square (χ2) | P value | |
|---|---|---|---|---|
| Il-23 expression (150 cases) | ||||
| Low | 37 (49.3%) | 24 (32%) | 4.669 | 0.030* |
| High | 38 (50.7%) | 51 (68%) | ||
| IL-28 expression (150 cases) | ||||
| Low | 36 (48%) | 26 (34.7%) | 2.749 | 0.097 |
| High | 39 (52%) | 49 (65.3%) |
NMA, non-mucinous adenocarcinoma; MA, mucinous adenocarcinoma; *P value ≤0.05 is significant
Table 3.
Relation of IL-23 Expression to Selected Clinicopathological & Histological Parameters within NMA & MA Groups
| NMA | MA | |||
|---|---|---|---|---|
| IL-23 Low expression (N=37) No. (%) | IL-23 High expression (N=38) No. (%) | IL-23 Low expression (N=24) No. (%) | IL-23 High expression (N=51) No. (%) | |
| Lymphovascular emboli | ||||
| Negative | 14 (37.8 %) | 16 (42.1 %) | 11 (45.8 %) | 12 (23.5 %) |
| Positive | 23 (62.2 %) | 22 (57.9 %) | 13 (54.2 %) | 39 (76.5 %) |
| P value | 0.82 | 0.05* | ||
| Chi-square (χ2) | 0.14 | 3.81 | ||
| Depth of invasion | ||||
| T1/T2 | 11 (29.7 %) | 7 (18.4 %) | 5 (20.8 %) | 1 (2.0 %) |
| T3/T4 | 26 (70.3 %) | 31 (81.6 %) | 19 (79.2 %) | 50 (98.0 %) |
| P value | 0.28 | FE 0.01* | ||
| Chi-square (χ2) | 131 | 7.89 | ||
| LN | ||||
| N0 | 19 (51.4 %) | 19 (50.0 %) | 13 (54.2 %) | 14 (27.5 %) |
| N1 | 11 (29.7 %) | 14 (36.8 %) | 5 (20.8 %) | 14 (27.5 %) |
| N2 | 7 (18.9 %) | 5 (13.2 %) | 6 (25.0 %) | 23 (45.0 %) |
| P value | 0.74 | 0.07 | ||
| Chi-square (χ2) | 0.68 | 5.22 | ||
| TNM stage | ||||
| I/II | 19 (51.4 %) | 18 (47.4 %) | 13 (54.2 %) | 14 (27.5 %) |
| III/IV | 18 (48.6 %) | 20 (52.6 %) | 11 (45.8 %) | 37 (72.5 %) |
| P value | 0.81 | 0.03* | ||
| Chi-square (χ2) | 0.12 | 5.05 | ||
| IL-28 expression | ||||
| Low | 21 (56.8 %) | 15 (39.5 %) | 17 (70.8 %) | 9 (17.6 %) |
| High | 16 (43.2 %) | 23 (60.5 %) | 7 (29.2 %) | 42 (82.4 %) |
| P value | 0.16 | <0.001** | ||
| Chi-square (χ2) | 2.24 | 20.38 | ||
NMA, non-mucinous adenocarcinoma; MA, mucinous adenocarcinoma; *P≤0.05 is significant **P <0.001 is highly significant; FE, Fisher’s Exact test
Table 4.
Relation of IL-28 Expression to Selected Clinicopathological & Histological Parameters within NMA & MA Groups
| NMA | MA | |||
|---|---|---|---|---|
| IL-28 Low expression (N=36) No. (%) | IL-28 High expression (N=39) No. (%) | IL-28 Low expression (N=26) No. (%) | IL-28 High expression (N=49) No. (%) | |
| Lymphovascular emboli | ||||
| Negative | 15 (41.7 %) | 15 (38.5 %) | 11 (42.3 %) | 12 (24.5 %) |
| Positive | 21 (58.3 %) | 24 (61.5 %) | 15 (57.7 %) | 37 (75.5 %) |
| P value | 0.82 | 0.12 | ||
| Chi-square (χ2) | 0.08 | 2.5 | ||
| Depth of invasion | ||||
| T1/T2 | 8 (22.2 %) | 10 (25.6 %) | 6 (23.1 %) | 0 (0.0 %) |
| T3/T4 | 28 (77.8 %) | 29 (74.4 %) | 20 (76.9 %) | 49 (100 %) |
| P value | 0.79 | FE 0.001* | ||
| Chi-square (χ2) | 0.12 | 12.29 | ||
| LN | ||||
| N0 | 17 (47.2 %) | 21 (53.8 %) | 14 (53.8 %) | 13 (26.5 %) |
| N1 | 13 (36.1 %) | 12 (30.8 %) | 8 (30.8 %) | 11 (22.4 %) |
| N2 | 6 (16.7 %) | 6 (15.4 %) | 4 (15.4 %) | 25 (51.0 %) |
| P value | 0.86 | 0.008* | ||
| Chi-square (χ2) | 0.34 | 9.56 | ||
| TNM stage | ||||
| I/II | 16 (44.4 %) | 21 (53.8 %) | 14 (53.8 %) | 13 (26.5 %) |
| III/IV | 20 (55.6 %) | 18 (46.2 %) | 12 (46.2 %) | 36 (73.5 %) |
| P value | 0.49 | 0.02* | ||
| Chi-square (χ2) | 0.66 | 5.5 | ||
NMA, non-mucinous adenocarcinoma; MA, mucinous adenocarcinoma; *P≤0.05 is significant; FE, Fisher’s Exact test
To investigate the effect of IL-23 and IL-28 expression on prognosis, we analyzed differences in DFS and OS between high and low IL-23 and IL-28 expression within each group. In the non-mucinous group, the Kaplan-Meier test revealed no significant impact of IL-23 or IL-28 expression on DFS or OS (p = 0.39, log-rank [Mantel-Cox] = 0.73; p = 0.38, log-rank [Mantel-Cox] = 0.76; p = 0.88, log-rank [Mantel-Cox] = 0.023; and p = 0.78, log-rank [Mantel-Cox] = 0.024, respectively). In contrast, within the mucinous group, cases with high IL-23 and IL-28 expression showed significantly shorter DFS than those with low expression (p = 0.018, log-rank [Mantel-Cox] = 5.57 and p = 0.009, log-rank [Mantel-Cox] = 6.91, respectively). Additionally, cases with high IL-23 and IL-28 expression exhibited significantly shorter OS compared to those with low expression (p = 0.009, log-rank [Mantel-Cox] = 6.89 and p = 0.021, log-rank [Mantel-Cox] = 5.36, respectively) (Figures 3-6).
Figure 3.
The Overall Survival Function in Patients from NMA & MA Groups in Relation to IL-23 Expression in Tumor Cells
Figure 6.
The Disease-Free Survival Function in Patients from NMA and MA Groups in Relation to IL-28 Expression in Tumor Cells
Discussion
Preclinical studies using mice and human cell lines in models of sporadic and inflammation-related CRC have demonstrated a critical role of IL-23 and its downstream interactor IL-22 in CRC development and dissemination [28-31]. One study examining IL-10 signaling genes in matched neoplastic and non-neoplastic tissue pairs from CRC patients reported upregulation of IL-10R2 and STAT3 in CRC tissue. IL-22 signals through the upregulated IL-10R2 to activate STAT3, thereby contributing to the development of colorectal tumors. In addition to IL-10 and IL-22, IL-28 shares this receptor subunit [32]. Furthermore, IL-28 exhibits a high degree of structural similarity to IL-22 [36]. Given the close relationship between IL-22 and IL-28 and their shared use of the IL-10R2 subunit, we speculated that IL-23 and IL-28 may be interrelated in colorectal tumorigenesis. Due to the distinct molecular characteristics of MA and NMA, the present study hypothesized that this interrelation may differ between the two types.
The results of the present investigation demonstrated, for the first time, that high expression of IL-23 and IL-28 is more frequent in MA than in NMA; however, this difference was statistically significant only for IL-23 expression. Moreover, the current findings revealed a highly significant association between high expression of IL-23 and IL-28 within the MA group. These observations may be explained by the higher prevalence of KRAS mutations in MA [37] and the role of oncogenic KRAS in activating NF-κB signaling, which, in turn, enhances the transcription of various cytokines and chemokines essential for the initiation and progression of inflammation-induced cancer [38-40]. It has been observed that most CRC cases with a high frequency of KRAS mutations are associated with chronic inflammatory diseases [41] and that MA is more frequently diagnosed in patients with IBD [42]. This hypothesis is supported by a study conducted by Petanidis et al. [43], which demonstrated a significant correlation between high IL-23 expression and KRAS mutation in CRC patients.
When evaluating IL-23 expression with clinicopathological parameters and prognosis within each group, none of the examined parameters showed a significant association with IL-23 expression in the NMA group. In contrast, within the MA group, a significant association was observed between high IL-23 expression and lymphovascular invasion, advanced depth of invasion, late TNM stage, and shorter DFS and OS. This observation is consistent with previously reported progressive increases in IL-23 expression from non-neoplastic to neoplastic tissues [28, 44] and the established pro-oncogenic role of IL-23, which is mediated by the activation of STAT3 and leads to the upregulation of MMPs, VEGF, and Bcl-x, thereby promoting tumor survival and metastasis [45]. Similarly, Hu et al. [46] reported a significant association between high IL-23 expression in CRC and increased depth of invasion, advanced TNM stage, and shorter DFS and OS. Furthermore, the current findings align with those of Petanidis et al. [43], who found a significant relationship between IL-23 expression and KRAS-linked, stage-specific overexpression in CRC .
Conversely, Helbling et al. [47] found no significant association between IL-23 expression in CRC and clinicopathological characteristics or outcomes. They classified cases into negative and positive groups based on a threshold value for the average percentage of positive neoplastic cells [47]. In contrast, the present study incorporated both the intensity and percentage of stained tumor cells in assessing IL-23 expression. Variations in IL-23 staining intensity may help distinguish between different stages of CRC and their outcomes.
Regarding the relationship between IL-28 expression and the clinicopathological aspects and prognosis within each group, the results of the current study showed no significant association between IL-28 expression and any of the examined parameters within the NMA group. In contrast, within the MA group, our results revealed a significant association between high IL-28 expression and advanced pT stage, nodal spread, late TNM stage, and decreased DFS and OS. This finding is consistent with that of Sakahara et al. (2019), who demonstrated that activating mutations of KRAS contribute to tumorigenesis and drug resistance in CRC through upregulation of IFN/STAT signaling [48]. Additionally, it has been shown that the tumor suppressor gene BMP5 exerts its inhibitory effect in CRC by suppressing IL-28A expression, thereby blocking the JAK-STAT pathway [49]. Moreover, the present results align with those of Wang et al. (2023), who conducted a comprehensive analysis of IFN-λ genes across different malignancies, including colorectal adenocarcinoma, and concluded that endogenous IFN-λ expression is associated with poor prognosis, with IFN-λ2 and IFN-λ3 serving as independent prognostic factors. They further demonstrated that IFN-λ cooperates with STAT1, STAT2, and STAT3 to activate the JAK-STAT signaling pathway by upregulating the expression of IL24, IL10, IL26, and IL10RB, thereby promoting tumor progression [50].
In contrast to our findings, anti-proliferative effects of IFN-λ have been reported in colorectal adenocarcinoma models [51]. Moreover, Cheng et al. [52] demonstrated that IL-28B significantly inhibits CRC progression by suppressing the polarization of M2 macrophages .
The demonstrated pro- and anti-tumorigenic effects of IFN-λ in various CRC studies can be attributed to its emerging dual role in carcinogenesis. Several factors have been identified as regulators of IFN-λ function in cancer. One such factor is the level and duration of IFN-λ signaling in the tumor microenvironment. While low levels and short duration induce acute inflammation and tumor cell eradication, high levels and prolonged signaling result in chronic inflammation and inflammation-associated cancer [53]. Another factor involves the differential effects of IFN-λ on various immune cell types; IFN-λ can modulate the activity of both immune-stimulating and immune-suppressive cells [54]. Additionally, the type of interferon-stimulated genes (ISGs) activated may influence the effect of IFN-λ on cancer immunity. Some ISGs encode molecules that enhance immune responses, whereas others encode molecules that contribute to immune checkpoint blockade and immune suppression [53].
In conclusion, expressions of both IL-23 and IL-28 showed a highly significant interrelation in MA patients, suggesting a potential interplay between them. High expressions of IL-23 and IL-28 may have adverse prognostic implications for survival in MA. Further molecular studies are necessary to elucidate the interaction between IL-23 and IL-28 in colorectal tumorigenesis.
Figure 4.
The Overall Survival Function in Patients from NMA & MA Groups in Relation to IL-28 Expression in Tumor Cells
Figure 5.
The Disease-Free Survival Function in Patient from NMA and MA Groups in Relation to IL-23 Expression in Tumor Cells
Acknowledgements
Funding statement
This research did not receive any specific grant from funding agencies in the public, commercial or not-for-profit sectors.
Data Availability Statement
The data that support the findings of this study are included in this published article & available on reasonable request from the corresponding author
Ethical Approval Statement
This research was conducted in accordance with the ethical guidelines of the Research Ethics Committee of the Faculty of Medicine for Girls, Al-Azhar University, and in adherence to the 1964 Declaration of Helsinki and its subsequent amendments. Ethical approval was obtained from the Research Ethics Committee of the Faculty of Medicine for Girls, Cairo, Al-Azhar University, under approval number (IRB: 2023112161), dated 12-11-2024.
Disclosure Statement
The authors declare that they have no conflict of interes.
Abbreviations
Interleukin (IL), Colorectal Cancer (CRC), Mucinous Adenocarcinoma (MA), Non-Mucinous Adenocarcinoma (NMA), pathological tumor (pT), pathological node (pN), Tumor-Node-Metastasis (TNM), T helper 17 (Th17), Inflammatory Bowel Disease (IBD), Matrix Metalloproteinase 9 (MMP 9), Interferon (IFN), Tissue microarray (TMA), Lymph Node (LN), Ethylenediaminetetraacetic acid (EDTA), Immunohistochemical Score (IHS), Disease-free survival (DFS), Overall survival (OS), Signal Transducer & Activator of Transcription (STAT), Kirsten Rat Sarcoma (KRAS), Nuclear Factor Kappa Beta (NF-kB), Bone Morphogenetic Protein 5 (BMP5), Janus kinase (JAk), Interferon stimulated genes (ISGs).
Author Contribution Statement
Study design: Abd Al-rahman Mohammad Foda; Data interpretation: Abd Al-rahman Mohammad Foda, Azza kamal Taha & Rania Refat Abdel Maqsoud; Drafting of the manuscript: Azza kamal Taha, Doaa E.A. Salama, Amira Nasr Ismail Elsokary & Rania Refat Abdel Maqsoud; Final revision of the manuscript: Abd Al-rahman Mohammad Foda, Doaa E.A. Salama, Amira Nasr Ismail Elsokary, Azza kamal Taha & Rania Refat Abdel Maqsoud.
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Associated Data
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Data Availability Statement
The data that support the findings of this study are included in this published article & available on reasonable request from the corresponding author