miR-24-3p靶向KLF6基因调控IL-6/STAT3信号通路影响食管癌细胞的活力和凋亡

miR-24-3p targeting KLF6 gene affects viability and apoptosis of esophageal cancer cells via IL-6/STAT3 signaling pathway

目的:探讨微小RNA-24-3p(miR-24-3p)对食管癌细胞活力和凋亡的影响及机制。方法:以人正常食管上皮细胞HEEC为对照,采用RT-qPCR检测食管癌细胞TE11、Eca109和EC9706中miR-24-3p和KLF6 mRNA的表达,Western blot检测KLF6蛋白的表达。用anti-miR-24-3p和KLF6 siRNA转染EC9706细胞,MTT检测细胞活力,流式细胞术检测细胞凋亡率,Western blot检测检测细胞中与增殖、凋亡相关的蛋白以及IL-6/STAT3信号通路相关蛋白的表达,ELISA法检测IL-6的表达。双萤光素酶报告基因实验验证miR-24-3p与KLF6靶向调控的关系。结果:食管癌癌细胞TE11、Eca109和EC9706中miR-24-3p表达上调(P<0.05),KLF6的mRNA和蛋白表达下调(P<0.05)。敲减EC9706细胞miR-24-3p表达可抑制其细胞活力,诱导其凋亡,并抑制细胞CDK4、cyclin D1、CDC25A、p-STAT3、IL-6及Bcl-2的表达,促进caspase-3和Bax的表达。结论:miR-24-3p可靶向KLF6基因调控IL-6/STAT3信号通路影响食管癌细胞的生长和凋亡。

AIM: To investigate the effect of microRNA-24-3 p(miR-24-3 p) on the viability and apoptosis of esophageal cancer cells. METHODS: The expression of miR-24-3 p and KLF6 mRNA in the esophageal cancer cells TE11, Eca109 and EC9706 were detected by RT-qPCR. The protein expression of KLF6 was determined by Western blot. EC9706 cells were transfected with anti-miR-24-3 p and KLF6 siRNA. The cell viability was measured by MTT assay, the apoptotic rate was analyzed by flow cytometry, and the proliferation, apoptosis and IL-6/STAT3 signaling pathways related proteins were determined by Western blot. The level of IL-6 was measured by ELISA. The dual luciferase reporter gene assay was used to verify the relationship between miR-24-3 p and KLF6. RESULTS: The levels of miR-24-3 p were up-regulated in the esophageal cancer cells TE11, Eca109 and EC9706(P<0.05), and the expression of KLF6 at mRNA and protein levels was down-regulated(P<0.05). Knock-down of miR-24-3 p expression inhibited the cell viability, induced apoptosis, and inhibited the protein levels of CDK4, cyclin D1, CDC25 A, p-STAT3, Bcl-2 and IL-6, and promoted the protein expression of caspase-3 and Bax in EC9706 cells. CONCLUSION: miR-24-3 p targets KLF6 gene to affect the viability and apoptosis of esophageal cancer cells by regulating IL-6/STAT3 signaling pathway.