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Published on: March 14, 2019
CHRM3-Associated miRNAs May Play a Role in Bile Acid-Induced Proliferation of H508 Colon Cancer Cells
Çağdaş Aktan1, Fatih Tekin2, Nevin Oruç2
1Department of Medical Biology, Beykent University Faculty of Medicine, İstanbul, Turkey.
Background:
It was well defined that proliferative effects of bile acids on colon epithelium are through interaction with muscarinic-3 receptors. Recently, microRNA emerged as an important regulator of gene expression and has been implicated in pathogenesis of many malignancies. However, the interaction of CHRM3 and microRNAs and their potential effects on colon carcinogenesis remains to be elucidated.
Methods:
In the current study, analysis of cell proliferation for 6 days after treatment with sodium taurolithocholate was analyzed by using WST-1 method. microRNAs which possibly target CHRM3 were identified by in silico analyses. Expression profiling of these microRNAs, expression changes of CHRM3 gene at mRNA level for H508 and SNU-C4 colon cancer cells were analyzed by quantitative polymerase chain reaction; the protein level of CHRM3 was analyzed using Western blot; apoptotic experiments were analyzed using the Annexin V assay. The Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed using the miRPath v3.0.
Results:
It was found that the expression level of CHRM3 gene was 6.133 ± 0.698-fold in H508 cells compared with SNU-C4 cells (P =.004). Treatment of H508 cells with sodium taurolithocholate caused 1.34 ± 0.4156-fold change in the expression level of CHRM3 gene (P =.0448). No apoptotic changes were observed in both colon cancer cells after treatment with sodium taurolithocholate. Different expression changes were detected of hsa-miR-129-5p, hsa-miR-30c-5p, hsa-miR-224-5p, hsa-miR-30b-5p, hsa-miR-522-3p, and hsa-miR-1246. Finally, hsa-miR-1246 and hsa-miR-522-3p could play a critical role in tumor development via bile acid-related genes in colon cancer.
Conclusion:
These findings reflected that CHRM3-dependent oncogenetic pathways might be in charge of colon cancer. We suggest that the microRNA expression profile of each individual colon cancer tissue is a unique digital signature.
Insights
Bile acids promote colon cancer via muscarinic-3 receptor (CHRM3) interactions. Specific microRNAs, hsa-miR-1246 and hsa-miR-522-3p, are key regulators in bile acid-related colon cancer development.
Area of Science:
- Gastroenterology and Hepatology
- Oncology
- Molecular Biology
Background:
- Bile acids are known to stimulate colon epithelial proliferation through muscarinic-3 receptors (CHRM3).
- MicroRNAs (miRNAs) are critical gene regulators implicated in various cancers, but their role in CHRM3-mediated colon carcinogenesis is unclear.
Purpose of the Study:
- To investigate the interaction between CHRM3 and microRNAs in the context of colon cancer.
- To explore the potential effects of these interactions on colon carcinogenesis.
Main Methods:
- Cell proliferation was assessed using the WST-1 assay after sodium taurolithocholate treatment.
- In silico analysis identified potential CHRM3-targeting miRNAs. Expression profiling of miRNAs, CHRM3 mRNA, and protein levels were analyzed via qPCR and Western blot.
- Apoptosis was evaluated using the Annexin V assay, and pathway enrichment analysis was performed using miRPath v3.0.
Main Results:
- CHRM3 gene expression was significantly higher in H508 cells compared to SNU-C4 cells.
- Sodium taurolithocholate treatment increased CHRM3 gene expression in H508 cells, but no apoptotic changes were observed.
- Specific miRNAs, including hsa-miR-1246 and hsa-miR-522-3p, showed differential expression and were identified as potentially critical in colon tumor development via bile acid-related pathways.
Conclusions:
- CHRM3-dependent pathways are implicated in colon cancer development.
- The unique microRNA expression profile of colon cancer tissue may serve as a diagnostic signature.
