miRNA26b靶向IL6／STAT3信号通路诱导肝癌细胞自噬的实验研究#br#

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摘要/Abstract

摘要： 目的探讨miRNA26b（miR26b）在肝癌细胞自噬调控中的作用及可能分子机制。
方法采用荧光定量PCR（QPCR）检测人正常肝细胞系HL7702和人肝癌细胞系HepG2、Hep3B、Bel7402及SSMC7721中miRNA26b的表达情况。双荧光素酶基因报告实验评价miR26b对IL6的靶向调控作用。向HepG2细胞转染miR26b模拟物 mimics（转染组）和阴性对照NC（阴性对照组），Western blotting检测两组IL6／STAT3信号通路以及自噬相关蛋白的表达。向HepG2细胞转染IL6 siRNA1和IL6 siRNA2，Western blotting 检测干扰IL6后STAT3激活及自噬相关蛋白表达。
结果HepG2、Hep3B、Bel7402细胞系中miR26b表达水平分别为034±0063、069±0092、057±0086，显著低于正常肝细胞系HL7702的115±0189，差异具有统计学意义（P＜005）。miR26b可抑制野生型IL6 3’UTR报告基因载体的荧光素酶活性，而对突变型IL6 3’UTRMUT的荧光素酶活性无影响。转染miR26b mimics后IL6、pSTAT3相对表达量显著降低（P＜005），诱导自噬相关蛋白Beclin1相对表达量和LC3Ⅱ／LC3Ⅰ比值显著升高（P＜005）。沉默IL6导致pSTAT3相对表达量显著降低（P＜005），诱导自噬相关蛋白Beclin1相对表达量和LC3Ⅱ／LC3Ⅰ比值显著升高（P＜005）。
结论miRNA26b靶向抑制IL6／STAT3信号通路的激活，进而诱导肝癌细胞自噬发生。

关键词: 肝细胞癌, miRNA26b, IL6／STAT3, 自噬

Abstract: ObjectiveTo investigate the role and possible molecular mechanism of miRNA26b （miR26b） in the regulation of autophagy in hepatocellular carcinoma cells.
MethodsThe expression of miRNA26b in human normal liver cell line HL7702 and human hepatoma cell lines HepG2， Hep3B， Bel7402 and SSMC7721 was detected by fluorescence quantitative PCR （QPCR）. Dual luciferase reporter assay was used to evaluate the regulatory effect of miR26b on IL6. MiR26b mimics （transfected group） and negative control NC （negative control group） were transfected into HepG2 cells， and Western blotting was used to detect the expression of IL6／STAT3 signaling pathway and autophagy related protein in two groups. IL6 siRNA1 and IL6 siRNA2 were transfected into HepG2 cells and Western blotting was used to detect STAT3 activation and autophagy related protein expression after interfering IL6.
ResultsThe expression levels of miR26b in HepG2， Hep3B and Bel7402 cell lines were 034±0063， 069±0092， 057±0086， which was significantly lower than that of normal liver cell line HL7702 （115±0189）， and the difference was statistically significant （P＜005）. MiR26b inhibited the luciferase activity of wildtype IL6 3’UTR reporter gene vector， but had no effect on the luciferase activity of mutant IL6 3’ UTRMUT. After transfection of miR26b mimics， the relative expression of IL6 and pSTAT3 decreased significantly （P＜005）， and the relative expression of autophagy related protein Beclin1 and the ratio of LC3 II／LC3 I increased significantly （P＜005）. After the silence IL6 expression， the relative expression of pSTAT3 decreased significantly （P＜005）， and the relative expression of autophagy related protein Beclin1 and the ratio of LC3 II／LC3 I increased significantly （P＜005）.
ConclusionMiRNA26b can inhibit the activation of IL6／STAT3 signaling pathway and induce autophagy in hepatoma cells.

Key words: Hepatocellular carcinoma, miRNA26b, IL6／STAT3, Autophagy

夏婷, 胡衡, 朱群安熊勰. miRNA26b靶向IL6／STAT3信号通路诱导肝癌细胞自噬的实验研究#br#

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[J]. 临床肿瘤学杂志, 2018, 23(7): 593-598.

XIA Ting, HU Heng, ZHU Qunan, XIONG Xie.. Experimental study of miRNA26b targeting IL6／STAT3 signaling pathway induces autophagy in hepatocellular carcinoma cells#br#

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[J]. Chinese Clinical Oncology, 2018, 23(7): 593-598.

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