miR-556-3p通过调控PTEN/AKT信号通路影响肝细胞癌的转移活性  

MiR-556-3p affects the metastatic activity of HepG2 cells by regulating PTEN/AKT signaling pathway in vitro

作　　者：仲金龙 施琳[1] Zhong Jinlong;Shi Lin(Department of Pathology,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010059,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古医科大学附属医院病理科,呼和浩特市010059

出　　处：《实用肝脏病杂志》2023年第6期781-784,共4页Journal of Practical Hepatology

基　　金：内蒙古自治区卫生健康委员会科技计划项目(编号:202201204)。

摘　　要：目的通过探索miR-556-3p在肝癌细胞发生发展过程中的作用机制,以期为肝细胞癌的预防和治疗提供一个新思路。方法取肝细胞癌组织和癌旁肝组织,采用PCR法检测miR-556-3p水平,采用免疫组化法检测磷酸酯酶与张力蛋白同源物(PTEN)表达,采用荧光素酶法检测细胞作用靶点,应用miR-556-3p mimic、miR-556-3p NC、PTEN cDNA和PTEN siRNA转染HepG2细胞,采用Western blotting法检测PTEN/AKT信号通路相关蛋白表达。采用CCK-8法检测细胞增殖能力,采用Transwell法检测细胞侵袭力,使用流式细胞术检测细胞凋亡。应用生物信息学法预测miR-556-3p对HepG2细胞PTEN/AKT通路的调控作用。结果肝癌组织miR-556-3p水平显著低于癌旁组织(P<0.05),HepG2细胞miR-556-3p水平显著低于正常肝细胞(P<0.05);荧光素酶分析显示,PTEN是miR-556-3p的直接靶点;miR-556-3p处理的HepG2细胞增殖、侵袭和凋亡率显著低于miR NC处理组(P<0.05);HepG2细胞PTEN过表达逆转了miR-556-3p对细胞生长抑制和凋亡诱导作用;miR-556-3p通过靶向PTEN抑制了HepG2细胞PTEN/AKT的激活。结论肝癌组织miR-556-3p水平降低,而miR-556-3p可抑制肝癌细胞增殖、侵袭并诱导细胞凋亡。肝癌细胞PTEN受miR-556-3p调控,果提示miR-556-3p与肝癌的恶性发展密切相关。Objective This study aimed to explore the mechanism of miR-556-3p in the development of hepatocellular carcinogenesis.Methods The miR-556-3p level in cancerous and its adjacent non-cancerous tissues from patients with hepatocellular carcinoma was detected by real-time fluorescence quantitative PCR,and the phosphatase and tensin homolog(PTEN)expression was revealed by immunohistochemistry.The expression patterns of PTEN/protein kinase B(Akt)signaling pathway-related proteins were detected by Western blotting after transfection of HepG2 cells with miR-556-3p mimic,miR-556-3p NC,PTEN cDNA and PTEN siRNA.The proliferation ability of HepG2 cells was studied by CCK-8 method,the invasion ability of cells was determined by Transwell method,and the apoptosis was detected by flow cytometry.The regulatory roles of miR-556-3p on PTEN/AKT pathway in HepG2 cells were evaluated by bioinformatics.Results The miR-556-3p loads in cancerous tissues was significantly lower than that in para-neoplastic tissues(P<0.05),and the miR-556-3p loads in HepG2 cells was also significantly lower than that in LO_(2) cells(P<0.05);the luciferase analysis showed that PTEN was a direct target of miR-556-3p;the proliferation rate,invasion rate and apoptosis rate in miR-556-3p-transfected HepG2 cells decreased significantly than in miR NC-transfected cells(P<0.05);the PTEN overexpression reversed the growth inhibition and apoptosis induction of miR-556-3p in HepG2 cells;the miR-556-3p inhibited PTEN/AKT activation by targeting PTEN.Conclusions The miR-556-3p loads decrease in hepatocellular carcinoma tissues,and the miR-556-3p inhibits the proliferation and invasion,and induce apoptosis in HepG2 cells.The PTEN is regulated by miR-556-3p in HepG2 cells.These findings above suggest that miR-556-3p is closely related to the transformation of liver cells,which needs further verification.

关 键 词：肝细胞癌 HepG2细胞 miR-556-3p 磷酸酯酶与张力蛋白同源物/磷酸化蛋白激酶B 增殖 凋亡 转移 体外 

分 类 号：R735.7[医药卫生—肿瘤]