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miR-125a-5p regulates migration and invasion of gastric cancer cells by down[1]regulating BAG4 expressio
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Abstract:
Objective: To explore the molecular mechanism of miR-125a-5p suppressing the migration and invasion of gastric cancer cells by regulating expression of Bcl-2-associated athanogene 4 (BAG4) gene. Method: A total of 82 pairs of gastric cancer tissues and corresponding para-cancer tissues were obtained from gastric cancer patients who received curative surgery at the First Hospital of Lanzhou University during January 2014 to December 2015. Human gastric cancer cell lines (MGC803, BGC823, SGC7901, HGC27) and human gastric epithelium cell line (GES-1) were also collected for this study. Real-time fluorescent quantitative PCR (qPCR) method was used to detect the expression level of miR-125a-5p in gastric cancer tissues, para-cancer tissues and gastric cancer cell lines. miR-125a-5p mimics, miR-125a-5p inhibitor, si-BAG4 (siRNA-BAG4) and negative control plasmids were transiently transfected into gastric cancer cells, respectively. The effect of miR-125a-5p/BAG4 signaling axis on migratory and invasive ability of gastric cancer cells was determined by Wound healing assay and Transwell invasion assay, respectively. WB was used to detect BAG4 protein expression in gastric cancer cells. The targeted regulatory relationship between miR-125a-5p and BAG4 was determined by Luciferase reporter gene assay. Result: miR-125a-5p was down-regulated in gastric cancer tissues and gastric cancer cell lines. The expression level of miR-125a-5p was not correlated with gender (P=0.953), age (P=0.772), tumor location (P=0.867), histological grade (P=0.745) and tumor size (P=0.088) of gastric cancer patients, but significantly correlated with T stage (P=0.003), N stage (P=0.001), M stage (P=0.027) and TNM stage (P=0.035) in gastric cancer patients, and the differences were statistically significant. Low expression of miR-125a-5p was an independent risk factor for overall survival of gastric cancer patients. miR-125a-5p significantly inhibited the migration and invasion of gastric cancer cells (all P<0.01). Knockdown BAG4 could reverse the inhibitory effect of miR-125a-5p inhibitor on migration and invasion of gastric cancer cells. Luciferase reporter gene assay validated that miR[1]125a-5p could bind with BAG4 3'UTR (Untranslated Regions) to suppress its expression. Conclusion: miR-125a-5p inhibits the migration and invasion of gastric cancer cells by down-regulating the expression level of BAG4.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 82060527), Intra-hospital Fund Project of the First Hospital of Lanzhou University（ldyyyn2019-02）, and Science and Technology Development Guiding Plan Project of Lanzhou, China (No. 2020-ZD-66)
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