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. 2024 Dec 29;77(1):28. doi: 10.1007/s10616-024-00692-5

miR-141-3p inhibited BPA-induced proliferation and migration of lung cancer cells through PTGER4

Feng Ling 1,#, Wenbo Xie 2,#, Xiang Kui 3, Yuyin Cai 1, Meng He 1, Jianqiang Ma 1,✉
PMCID: PMC11683044  PMID: 39741890

Abstract

The chemical substance bisphenol A (BPA) is widely used in household products, and its effect on human health has frequently been the focus of research. The aim of this study was to explore the potential molecular regulatory mechanism of BPA on the proliferation and migration of lung cancer cells. In this study, the H1299 and A549 lung cancer cell lines were selected as the study objects. The cells were treated with different concentrations of BPA (0, 0.1, 1, or 10 μM), and cell viability, proliferation, and migration were evaluated by CCK-8, EdU, clonogenic, and scratch test assays. Western blotting and RT‒qPCR were used to detect the expression of related proteins and genes. Our findings indicated that BPA markedly enhanced both the proliferation and migration capacities of lung cancer cells. In BPA-treated lung cancer cells, the level of miR-141-3p was decreased, PTGER4 expression was significantly increased, and PTGER4 knockdown reduced BPA-induced lung cancer cell proliferation and migration. In addition, miR-141-3p can target and negatively regulate the expression of PTGER4 and further inhibit PI3K/AKT signaling pathway activation and MMPs expression. Moreover, PTGER4 overexpression weakened the inhibitory effect of the miR-141-3p mimic on the proliferation and migration of lung cancer cells. In conclusion, miR-141-3p can inhibit the proliferation and migration of BPA-induced lung cancer cells by downregulating PTGER4, providing a new potential target for the treatment and prevention of lung cancer.

Supplementary Information

The online version contains supplementary material available at 10.1007/s10616-024-00692-5.

Keywords: Lung cancer, Bisphenol A, PTGER4, MiR-141-3p, PI3K/AKT

Introduction

Lung cancer ranks as the foremost cause of cancer-related death globally and is considered the most aggressive type of malignant tumor (Harðardottir et al. 2022). Over the last three decades, there has been a steady rise in the rates of lung cancer, establishing it as the quickest escalating form of cancer and posing a substantial threat to human health and well-being Wang et al. 2024). There are several treatment options available, including surgery, radiotherapy, and systemic therapy (Li et al. 2022; Vinod 2020). Regrettably, the detection of lung cancer is frequently delayed, which adversely affects the efficacy of treatment (Shankar et al. 2019). The onset of lung cancer is influenced by various factors, such as smoking and genetics (Bade et al. 2020; Liu et al. 2021). Recent advancements in prevention, screening, treatment, and symptom management present new possibilities to reduce the burden of lung cancer (Gandhi et al. 2018). Nevertheless, owing to existing medical limitations, there is still room for improvement in the treatment and prognosis of lung cancer patients.

Bisphenol A (BPA) is a common endocrine-disrupting chemical (EDC) widely used in the manufacture of polycarbonate plastics and epoxy resins (Song et al. 2021a). The inadequate polymerization or hydrolysis of these polymers at high temperatures, under acidic conditions, or due to enzymatic activity results in the release of BPA from plastic materials (Almeida et al. 2018). Numerous studies have indicated a correlation between exposure to BPA and the onset and progression of cancers such as breast cancer, ovarian cancer, and prostate cancer (Lacouture et al. 2022; Ma et al. 2020; Samtani et al. 2018). A case‒control study also revealed that BPA levels in the urine of lung cancer patients are notably increased and closely associated with ER stress gene variations (Li et al. 2020). BPA, according to in vitro experiments, may stimulate the proliferation and metastasis of lung cancer by activating GPER/EGFR/ERK1/2 signaling pathways (Zhang et al. 2014). However, the molecular mechanism of BPA-induced lung cancer cell proliferation and migration is still not fully understood, and further studies are needed to explore this area.

PTGER4, a G protein-coupled receptor, serves as a mediator of the effects of prostaglandin E2. Found on chromosome 5p13.1, it codes for the EP4 subunit, which produces receptors for PGE2 (Gabbani et al. 2016). As a key player in various vital biological processes, PTGER4 is essential for the proliferation of cancer cells, renewal of stem cells, and development of blood vessels within tumors (Baba et al. 2010; Cui et al. 2021). Additionally, it regulates cell movement and immunity (Tongtawee et al. 2018; Zhong et al. 2021). Numerous studies have shown a significant correlation between PTGER4 expression and the progression of different types of cancers, especially colorectal cancer (Yu et al. 2023). PTGER4 expression is also more prevalent in lung cancer cells than in other types of cancer cells(De Paz Linares et al. 2021; Majumder et al. 2018). Non-small cell lung cancer (NSCLC) is particularly common (Nandi et al. 2017). It is currently unknown how PTGER4 affects lung cancer cell proliferation and migration caused by BPA.

In this study, we aimed to explore the molecular mechanism by which BPA induces lung cancer cell proliferation and migration. We found that miR-141-3p can inhibit BPA-induced lung cancer cell proliferation and migration through PTGER4. This discovery provides a new perspective for understanding the pathogenesis and development of lung cancer and provides potential targets for developing more effective lung cancer treatment strategies.

Materials and methods

Cell culture

The human lung cancer cell lines A549 (SCSP-503) and H1299 (TCHu160) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in F-12 K medium (#21,127,022, Gibco) or RPMI 1640 medium (11,875,093, Gibco) supplemented with 10% FBS or 1% penicillin‒streptomycin. The cells were cultured in an incubator at 37 °C with 5% CO2.

The A549 and H1299 cell lines were randomly grouped as follows: control group: cells without treatment; BPA group: the cells were cultured with different concentrations of BPA (0, 0.1, 1, or 10 µM) for 24 h (Lee et al. 2023; Song et al. 2021b); BPA + si-PTGER4 group: si-PTGER4 was transfected into cells and cultured with 10 µM BPA for 24 h; BPA + miR-141-3p mimic group: cells were transfected with the miR-141-3p mimic prior to being exposed to 10 µM BPA for 24 h; BPA + miR-141-3p mimic + OE-PTGER4 group: the miR-141-3p mimic and OE-PTGER4 were cotransfected into cells prior to being exposed to 10 µM BPA for 24 h. Transfection was performed via a Lipofectamine™ 2000 kit (11,668,027, Thermo). miR-141-3p was provided by Fenghui Biotechnology Co., Ltd., while si-PTGER4 and OE-PTGER4 were supplied by Shanghai Sangon (China).

CCK-8 assay

The viability of H1299 and A549 cells was evaluated via a CCK-8 assay. A total of 100 μL per well of each type of cell was seeded into 96-well plates, with varying concentrations of BPA added to each well (0, 0.1, 1, or 10 µM). After incubation for 48 h at 37 °C with 5% CO2 and 90% humidity, CCK-8 solution was added to each well. The viability of the cells was assessed via a Biotek microplate reader at a wavelength of 450 nm.

EdU assay

H1299 and A549 cells in logarithmic growth stages were inoculated in 24-well plates at a density of 1 × 104/well. After 24 h of culture, EdU working solution was added to the cells of each group for further incubation for 12 h, after which the cells were fixed with 4% paraformaldehyde for 30 min, and staining was performed as per the instructions provided in the EdU kit. (Beyotime, Shanghai, China). Finally, the stained cells were examined under a fluorescence microscope.

Colony formation experiment

After the H1299 and A549 cells in each group were digested, 500 cells/well were inoculated into 24-well plates. The culture media were changed every three to four days. Seventeen days postseeding, the cells were fixed with methanol and then stained for 15 min with crystal violet. Colony formation was assessed by capturing images with a microscope.

Scratch test

H1299 and A549 cells in each group were collected during the logarithmic growth phase, digested, and regularly counted. The cells were inoculated in 6-well plates at a density of 4 × 105/well. Once the cells reached a confluent monolayer, a sterile 10 μL pipette tip was used to create a scratch along a premarked line in the 6-well plate, removing any detached cells. Three to five regions were selected in each well for imaging to record the width of the scratch, and the cells were then incubated for 24 h. Finally, images were taken to monitor changes in the scratch width at the same locations, and these images were analyzed for comparative assessment.

RT‒qPCR

Total RNA was extracted according to the manufacturer's instructions. To synthesize cDNA, a SweScript RT I First-Strand miRNA cDNA Synthesis Kit (G3331-50, Servicebio, China) or an mRNA cDNA Synthesis Kit (18,091,050, Thermo Scientific, USA) was used. RT‒qPCR was performed with SYBR Green qPCR Master Mix (G3320-01, Servicebio, China). GAPDH and U6 were used as internal reference genes. The relative expression levels were calculated via the 2−ΔΔCt method. The specific sequences of primers used in the analysis can be found in Table 1.

Table 1.

Sequences of the primers

Genes Sequence (F: Forward primer, R: Reversed primer)
PTGER4 F: 5′-CCGGCGGTGATGTTCATCTT
R: 5′-CCCACATACCAGCGTGTAGAA
miR-141-3p F: 5′-GCGCGTAACACTGTCTGGTAA
R: 5′-AGTGCAGGGTCCGAGGTATT
miR-132-3p

F: 5′-GCGCGTAACAGTCTACAGCCA

R: 5′-AGTGCAGGGTCCGAGGTATT
U6

F: 5′-CTCGCTTCGGCAGCACA

R: 5′-AACGCTTCACGAATTTGCGT
GAPDH F: 5′-GGAGCGAGATCCCTCCAAAAT
R: 5′-GGCTGTTGTCATACTTCTCATGG
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Western blot

Total protein was isolated from each group using RIPA lysis buffer, and the protein concentration was determined with a BCA kit. PVDF membranes were used to separate the protein samples after SDS‒PAGE. The membranes were then blocked for 1 h at room temperature in 5% skim milk. The membranes were subsequently incubated with primary antibodies overnight at 4 °C. Anti-PTGER4 (1:1000, PA5-35,259, Invitrogen), anti-p-PI3K p85/p55 (Tyr458 and Tyr199) (1:1000, #PA5-17,387, Thermo Fisher, USA), anti-PI3K (1:1000, #710,400, Thermo Fisher, USA), anti-Akt (1:000, #9272, CST, USA), anti-p-Akt (Ser473) (1:1000, #9271, CST, USA), anti-mTOR (1:1000, #2972, CST, USA), anti-p-mTOR (Ser2448) (1:1000, #2971, CST, USA), anti-MMP1 (1:1000, ab134184, Abcam, UK), anti-MMP2 (1:1000, ab181286, Abcam, UK), and anti-MMP9 (1:1000, ab76003, Abcam, UK) were used as primary antibodies. Then, the specific HRP-linked secondary antibody (1:1000, #7074, CST, USA) was added for 1 h at room temperature. Finally, with an anti-GAPDH antibody (1:2000) as an internal reference protein, the proteins were visualized via the chemiluminescence method.

Dual-luciferase gene reporter assay

The database starbase.sysu.edu.cn was used to predict the binding sites between miR-141-3p and PTGER4. To examine this interaction, wild-type (WT) and mutant (MUT) PTGER4 plasmids were constructed and named PTGER4-WT and PTGER4-MUT, respectively. PTGER4-WT and PTGER4-MUT plasmids were cotransfected with the miR-NC or miR-141-3p mimic into 293 T cells. Following transfection for 48 h, the Dual-Luciferase Reporter Assay System (Promega, WI, USA) was used to measure the luciferase activity in the cells of the various experimental groups.

Statistical analysis

All the experiments were repeated three times. The data are expressed as the means ± standard deviations. Statistical analysis was performed using GraphPad Prism 8.0 software. Student's t test and one-way analysis of variance were used to assess differences between groups. Statistical significance was defined as a P value less than 0.05.

Results

BPA induced the proliferation and migration of lung cancer cells

A549 and H1299 cells were exposed to different concentrations of BPA, and the viability, proliferation, and migration of the cells were examined. The viability, proliferation rates, number of clones and migration ability of H1299 and A549 cells increased with increasing BPA concentration compared with those of the control groups treated with 0 μM BPA, and the effect of 10 µM BPA was the most obvious (Fig. 1A-D). The above experiments showed that BPA could induce the proliferation and migration of lung cancer cells in a dose-dependent manner. In subsequent experiments, the cells were treated with 10 µM BPA.

Fig. 1.

Fig. 1

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BPA induced the proliferation and migration of lung cancer cells

A: A CCK-8 assay was used to detect cell viability; B: An EdU assay was used to detect cell proliferation; C: Colony formation assay; D: Cell migration was detected by a scratch test. Compared with the control group, **P < 0.01, ***P < 0.001.

High expression of PTGER4 in BPA-induced lung cancer cells

Previous studies have shown that PTGER4 is overexpressed in various cancer cells, including lung cancer cells, and is closely related to the occurrence of non-small cell lung cancer (De Paz Linares et al. 2021). Next, PTGER4 expression in H1299 and A549 cells stimulated with BPA was detected. Compared with the control, BPA increased the expression levels of PTGER4 mRNA and protein in cells (Fig. 2A-B). These findings indicated that PTGER4 was highly expressed in BPA-induced lung cancer cells.

Fig. 2.

Fig. 2

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High expression of PTGER4 in BPA-induced lung cancer cells

A: PTGER4 mRNA expression was detected by RT‒qPCR; B: Western blot detection of PTGER4 protein expression. Compared with the control group, **P < 0.01, ***P < 0.001.

Downregulation of PTGER4 inhibited BPA-induced proliferation and migration of lung cancer cells

Next, we explored the effects of PTGER4 on BPA-induced lung cancer cell proliferation and migration. After the cells were transfected with si-PTGER4, western blot analysis was performed to assess the transfection efficiency. The results revealed that PTGER4 expression levels were significantly reduced after PTGER4 was knocked down (Fig. 3A). Next, the effects on cell viability, proliferation, migration, and colony formation were assessed. Compared with control cells, lung cancer cells treated with BPA presented increased viability, proliferation, migration, and colony formation. However, transfection with si-PTGER4 inhibited the ability of BPA to promote the proliferation and migration of lung cancer cells (Fig. 3B-E). These results indicated that downregulating PTGER4 inhibited the proliferation and migration of BPA-induced lung cancer cells.

Fig. 3.

Fig. 3

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Downregulation of PTGER4 inhibits BPA-induced proliferation and migration of lung cancer cells

A: si-PTGER4 transfection efficiency was detected by western blot. B: Cell viability was determined via the CCK-8 assay. C: An EdU assay was used to detect cell proliferation. D: Colony formation was detected via the colony formation assay. E: Cell migration was detected via the scratch test. Compared with the control group, ***P < 0.001; compared with the BAP group, ###P < 0.001.

miR-141-3p targeted PTGER4 expression

The miRNA genes associated with lung cancer were screened via the GSE194300 dataset (Fig. 4A). Through R language analysis of upstream PTGER4 genes and intersection, the results revealed that miR-132-3p and miR-141-3p were underexpressed in the lung cancer datasets and were upstream PTGER4 target genes (Fig. 4B). The expression levels of miR-141-3p and miR-132-3p in the cells were then assessed. Compared with those in the control group, miR-141-3p and miR-132-3p were significantly downregulated in BPA-induced cells, and the degree of downregulation of miR-141-3p was greater (Fig. 4C-D). Next, the website starbase.sysu.edu.cn/ was used to predict that there was a targeted binding site between miR-141-3p and PTGER4 (Fig. 4E). A dual-luciferase reporter assay was used to verify the targeting relationship between these genes. According to the findings of this investigation, the luciferase activity of PTGER4-WT was reduced by the miR-141-3p mimic, whereas no change was observed in PTGER4-MUT (Fig. 4F). Additionally, the cells were transfected with miR-141-3p mimics and inhibitors. The group that received the mimics presented increased miR-141-3p levels, whereas the group that received the inhibitors presented reduced miR-141-3p levels (Fig. 4G). A subsequent evaluation of PTGER4 mRNA and protein levels within the cells revealed a reduction in PTGER4 expression in the miR-141-3p mimic group, along with an increase in PTGER4 expression following the inhibition of miR-141-3p (Fig. 4H-I). These results indicate that miR-141-3p can negatively regulate the expression of PTGER4.

Fig. 4.

Fig. 4

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miR-141-3p targeted PTGER4 expression

A: Analysis of miRNAs in lung cancer samples using bioinformatics; B: Venn diagram; C-D: RT‒qPCR detection of miR-141-3p and miR-132-3p expression; E: Prediction of miR-141-3p and PTGER4 binding sites in a database; F: Dual-luciferase gene reporter assay; G: RT‒qPCR detection of miR-141-3p expression; H: RT‒qPCR detection of PTGER4 mRNA expression; I: PTGER4 expression was detected via western blot. Compared with the control group, *P < 0.05, **P < 0.01, and ***P < 0.001.

miR-141-3p inhibited BPA-induced proliferation and migration of lung cancer cells through PTGER4

Next, we explored how miR-141-3p regulates the proliferation and migration of lung cancer cells through PTGER4. After OE-PTGER4 was introduced into the cells, PTGER4 protein expression levels were evaluated. The results indicated that the expression level of PTGER4 in cells was increased after PTGER4 was overexpressed (Fig. 5A), indicating successful transfection. Next, we examined the effects of the overexpression of miR-141-3p and PTGER4 on the proliferation and migration of lung cancer cells. The results revealed that the overexpression of miR-141-3p inhibited the proliferation and migration of lung cancer cells, whereas the overexpression of PTGER4 promoted the proliferation and migration of lung cancer cells (Figure S1A-D). Next, the effects of miR-141-3p and PTGER4 on the proliferation and migration of BPA-induced lung cancer cells were further examined. The results revealed that after transfection with the miR-141-3p mimic, there was a decrease in cell viability, proliferation rates, migration abilities, and the number of clones. Nevertheless, PTGER4 overexpression weakened the effects of the miR-141-3p mimic (Fig. 5B-E). These findings indicate that miR-141-3p hinders the proliferation and migration of BPA-induced lung cancer cells through PTGER4.

Fig. 5.

Fig. 5

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miR-141-3p inhibits BPA-induced proliferation and migration of lung cancer cells through PTGER4

A: Western blot detection of OE-PTGER4 transfection efficiency; B: Detection of cell viability via the CCK-8 assay; C: EdU detection of cell proliferation; D: Colony formation assay; E: Cell migration was detected by a scratch test. Compared with the control group, ***P < 0.001; compared with the BPA group, ###P < 0.001; compared with the BPA + miR mimic group, &&P < 0.01, &&&P < 0.001.

miR-141-3p inhibits the PI3K/AKT signaling pathway and MMPs expression by downregulating PTGER4

The PI3K/AKT signaling pathway plays a crucial role in the occurrence and metastasis of lung cancer; it is highly expressed in non-small cell lung cancer (Zou et al. 2024) and plays a key role in cell survival and abnormal activation during cancer development. For example, ILTPs inhibit the growth and proliferation of lung cancer A549 cells by inhibiting the activation of the PI3K/AKT signaling pathway (Chen et al. 2022; Wu et al. 2024). Furthermore, mTOR acts as a key downstream effector in this signaling pathway, contributing significantly to lung cancer progression. Therefore, we further investigated the effects of miR-141-3p on the PI3K/AKT signaling pathway in BPA-induced H1299 and A549 lung cancer cells. A western blot analysis was used to detect the expression levels of proteins related to the PI3K/AKT signaling pathway. Compared with those in the control group, the levels of p-PI3K, p-AKT and p-mTOR were increased after BPA treatment, which promoted the activation of the PI3K/AKT pathway. Furthermore, the overexpression of miR-141-3p inhibited the activation of the PI3K/AKT pathway, and the overexpression of PTGER4 alleviated the inhibitory effect of the miR-141-3p mimic (Fig. 6A). In addition, matrix metalloproteinases (MMPs) are important enzymes that lead to tumor cell migration, invasion and metastasis. High levels of MMPs are strongly associated with tumor growth and invasion, and the MMPs family members MMP-1, MMP-2, and MMP-9 are important indicators of lung cancer development (Kowalczyk et al. 2023; Shen et al. 2020). Thus, we examined the MMPs levels in the BPA-induced lung cancer cell lines H1299 and A549. BPA promoted the expression of MMP-1, MMP-2, and MMP-9, and the effects of BPA were weakened after transfection with the miR-141-3p mimic, whereas further overexpression of PTGER4 weakened the effects of the miR-141-3p mimic and increased the levels of MMP-1, MMP-2, and MMP-9 (Fig. 6B). The above results indicated that miR-141-3p inhibited the activation of the PI3K/AKT signaling pathway and the expression of MMPs by downregulating PTGER4.

Fig. 6.

Fig. 6

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miR-141-3p inhibits the PI3K/AKT signaling pathway and MMPs expression by downregulating PTGER4

A: Western blot analysis was performed to detect the expression of PI3K/AKT signaling pathway-related proteins. B: Western blotting was used to detect the protein expression of MMP-1, MMP-2 and MMP-9. Compared with the control group, ***P < 0.001; compared with the BPA group, ##P < 0.01, ###P < 0.001; compared with the BPA + miR mimic group, &P < 0.05, &&P < 0.01, &&&P < 0.001.

Discussion

BPA can bind to estrogen receptors in the body, affecting the production, transportation, and degradation of endogenous estrogen (Han et al. 2023). Increasingly, research indicates that BPA regulates gene expression and affects various cellular signaling pathways by activating specific receptors, such as PPARγ, Nrf2, and AhR (Cimmino et al. 2020; MacKay 2018). These effects can potentially lead to numerous health problems, including cancer development, reproductive toxicity, abnormal inflammation or immune responses, and abnormalities in brain or nervous system development (Murata 2018). Our research revealed that BPA enhanced lung cancer cell proliferation and migration in a concentration-dependent manner. Moreover, BPA activates the PI3K/AKT/mTOR pathway in the liver to initiate lipid biosynthesis (Han et al. 2018; Yecies et al. 2011). The chronic inflammation, oxidative stress, DNA damage, and fibrosis responses induced by BPA in multiple organs are closely associated with these mechanisms (Mansour et al. 2020). Numerous studies have highlighted the detrimental impact of BPA on lung tissues, highlighting the frequent involvement of inflammation, oxidative stress, and apoptosis in lung injuries caused by BPA (Hrenak 2020; Shi et al. 2023). In this study, BPA activated the PI3K/AKT pathway in H1299 and A549 cells, suggesting that the PI3K/AKT pathway is related to the progression of lung cancer.

MicroRNAs (miRNAs) are highly evolutionarily conserved endogenous RNAs that are approximately 22 nucleotides in length (Iqbal et al. 2019; Uddin 2018). miR-141-3p is a tumor suppressor gene. Previous studies have shown that the levels of miR-141-3p in NSCLC cells and tissues are significantly reduced (Li et al. 2019). Furthermore, miR-141-3p is thought to regulate the proliferation of NSCLC cells by controlling PHLPP1 and PHLPP2 (Huang et al. 2022). In this study, bioinformatics analysis revealed that miR-141-3p and miR-132-3p were significantly underexpressed in lung cancer tissues. Further experiments demonstrated that the downregulation effect of miR-141-3p was most obvious in BPA-induced lung cancer cells. Furthermore, miRNAs play a role in regulating multiple cancer-related biological processes by binding to the 3'-UTRs of target genes (Zhan et al. 2024). In this study, we found that miR-141-3p can target and negatively regulate the expression of PTGER4 and regulate the proliferation and migration of BPA-induced lung cancer cells by targeting downstream PTGER4 genes.

PTGER4 is involved in the regulation of many diseases, such as ulcerative colitis and lung cancer (Yu et al. 2023). Using RT‒qPCR and Western blotting techniques, our study confirmed increased PTGER4 expression in lung cancer cells stimulated with BPA. The important role of PTGER4's encoded receptor in prostaglandin signaling is highlighted in regard to conveying signals from the COX-2 enzyme and PGE2. This series of signals leads to an increase in factors that promote the proliferation, amplification, angiogenesis, migration, and invasion of colorectal cancer (CRC) cells. As a result, this receptor plays a significant role in the advancement and growth of colon tumors (Karpisheh et al. 2019). In addition, the ERK, PI3K/Akt and cAMP/PKA/CREB signaling pathways are activated, and the inhibition of cAMP-dependent signal transduction is a key mechanism in controlling tumor cell growth and metastasis, all of which are modulated by the expression level of the PTGER4 receptor (Karpisheh et al. 2020). Although the important role of PTGER4 in cancer has been extensively studied, the specific mechanism by which PTGER4 affects the proliferation and migration of lung cancer cells induced by BPA needs to be further explored. This study demonstrated that the downregulation of PTGER4 inhibited the proliferation and migration of BPA-induced lung cancer cells, and further experiments demonstrated that miR-141-3p inhibited the activation of the PI3K/AKT signaling pathway and MMPs expression through the downregulation of PTGER4, thus hindering the progression of BPA-induced lung cancer.

The PI3K/Akt pathway is a core signaling pathway that regulates cell functions such as survival, differentiation, and proliferation. In this pathway, the AKT protein is activated by PI3K through a series of regulators, and the subsequently activated AKT promotes the phosphorylation of downstream mTOR (Liu et al. 2018). Recent studies suggest that the PI3K/AKT pathway negatively influences apoptosis and autophagy in human cancers, especially in lung cancer, by phosphorylating mTOR (Gong et al. 2022). Our data revealed that BPA treatment significantly increased the protein levels of p-PI3K, p-AKT, and p-mTOR in lung cancer cells. Notably, the miR-141-3p mimic was able to mitigate this promoting effect of BPA. However, PTGER4 overexpression mitigated the effects of the miR-141-3p mimic and promoted the expression of PI3K/AKT pathway-related proteins. These findings indicate that BPA activates the PI3K/AKT pathway in lung cancer cells.

However, there are several limitations to the current study. First, we investigated the effects of the miR-141-3p/PTGER4 molecular axis on the proliferation and migration of lung cancer cells only at the cellular level. Owing to the complexity of the in vivo environment, further validation of this molecular mechanism in animal models is needed in the future. Second, the effect of miR-141-3p on the progression of lung cancer through the regulation of the PI3K/AKT signaling pathway needs to be further explored in the future. In conclusion, this study reveals the important role of miR-141-3p in the development of lung cancer. miR-141-3p inhibited the proliferation and migration of lung cancer cells induced by BPA by downregulating the expression of PTGER4. This discovery not only reveals a new mechanism for the development of lung cancer but also provides a potential target for the development of novel lung cancer treatment strategies.

Supplementary Information

Below is the link to the electronic supplementary material. Figure S1. Effects of miR-141-3p and PTGER4 on the proliferation and migration of lung cancer cells A: Cell viability was measured by a CCK-8 assay. B: An EdU assay was used to detect cell proliferation. C: Colony formation assay. D: Cell migration was detected by a scratch test. Compared with the control group, **P<0.01, ***P< 0.001.

Supplementary file1 (TIF 34590 KB) (33.8MB, tif)

Acknowledgements

Not applicable.

Author contributions

Conceptualization: Feng Ling, Wenbo Xie; methodology: Xiang Kui; software: Meng He; validation: Feng Ling, Jianqiang Ma; formal analysis: Wenbo Xie, Yuyin Cai; investigation: Feng Ling; resources: Jianqiang Ma; data curation: Xiang Kui; writing-original draft preparation: Feng Ling, Wenbo Xie; writing-review and editing: Jianqiang Ma; visualization: Yuyin Cai, Meng He; supervision: Wenbo Xie; funding acquisition: Jianqiang Ma. All authors have read and agreed to the published version of the manuscript.

Funding

This research was supported by the Yunnan Provincial Department of Science and Technology—Kunming Medical University Joint Special Basic Research Program (202201AY070001-120).

Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.

Declarations

Conflict of interest

The authors declare no competing interests.

Ethics approval and consent to participate

Not applicable.

Consent for publication

All the authors agreed on the publication of this manuscript.

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Feng Ling and Wenbo Xie contributed equally to this work.

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Associated Data

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

Supplementary Materials

Supplementary file1 (TIF 34590 KB) (33.8MB, tif)

Data Availability Statement

The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.

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