CHRM3 相关的 microRNAs 可能在胆汁酸诱导的 H508 结肠癌细胞增殖中发挥作用。
CHRM3-Associated miRNAs May Play a Role in Bile Acid-Induced Proliferation of H508 Colon Cancer Cells.
机构信息
Department of Medical Biology, Beykent University Faculty of Medicine, İstanbul, Turkey.
Department of Gastroenterology, Ege University Faculty of Medicine, İzmir, Turkey.
出版信息
Turk J Gastroenterol. 2023 Mar;34(3):298-307. doi: 10.5152/tjg.2022.22605.
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.
背景
胆汁酸对结肠上皮的增殖作用是通过与毒蕈碱-3 受体相互作用来实现的，这一点已经得到了很好的定义。最近，microRNA 作为一种重要的基因表达调控因子出现，并被认为与许多恶性肿瘤的发病机制有关。然而，CHRM3 与 microRNA 的相互作用及其对结肠癌发生的潜在影响仍有待阐明。
方法
在本研究中，通过 WST-1 法分析了用牛磺胆酸钠处理 6 天后的细胞增殖情况。通过计算机分析确定了可能靶向 CHRM3 的 microRNA。用定量聚合酶链反应分析 H508 和 SNU-C4 结肠癌细胞中这些 microRNA 的表达谱；用 Western blot 分析 CHRM3 基因的 mRNA 水平表达变化；用 Annexin V 测定法分析细胞凋亡实验。使用 miRPath v3.0 进行基因本体论和京都基因与基因组百科全书途径富集分析。
结果
发现 H508 细胞中 CHRM3 基因的表达水平与 SNU-C4 细胞相比为 6.133 ± 0.698 倍（P =.004）。H508 细胞用牛磺胆酸钠处理后，CHRM3 基因的表达水平变化了 1.34 ± 0.4156 倍（P =.0448）。用牛磺胆酸钠处理后，两种结肠癌细胞均未观察到凋亡变化。hsa-miR-129-5p、hsa-miR-30c-5p、hsa-miR-224-5p、hsa-miR-30b-5p、hsa-miR-522-3p 和 hsa-miR-1246 的表达变化不同。最后，hsa-miR-1246 和 hsa-miR-522-3p 可以通过与结肠癌相关基因相互作用，在肿瘤发生中发挥关键作用。
结论
这些发现反映了 CHRM3 依赖性致癌途径可能负责结肠癌。我们建议，每个个体结肠癌组织的 microRNA 表达谱是一个独特的数字特征。
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