整合素β4通过PI3K/AKT/mTOR促进HCC增殖和迁移  

Integrin β4 Promoting HCC Proliferation and Migration through the PI3K/AKT/mTOR Signaling Pathway

作　　者：唐小龙[1] 曹念蝶 宋雪翼 邵倩倩 TANG Xiaolong;CAO Niandie;SONG Xueyi;SHAO Qianqian(Medical College of Anhui University of Technology,Huainan Anhui 232001,China)

机构地区：[1]安徽理工大学医学院,安徽淮南232001

出　　处：《安徽理工大学学报（自然科学版）》2023年第4期94-102,共9页Journal of Anhui University of Science and Technology:Natural Science

基　　金：国家自然科学基金资助项目(82071862,81872017)。

摘　　要：目的 比较整合素β4(ITGB4)在不同肝癌细胞系和正常肝细胞中的表达差异,探讨ITGB4在肝细胞癌(HCC)中发挥的生物学效应以及调控其生物学效应的机制。方法 选取3种不同的肝癌细胞系SK-HEP-1、Huh7和HepG2以及正常肝细胞系L02,利用Western blot检测ITGB4在不同细胞系中的表达情况,并利用免疫荧光染色方法检测ITGB4在Huh7和HepG2之间的表达情况。选取低表达ITGB4的HepG2细胞和高表达ITGB4的Huh7细胞分别进行过表达ITGB4和RNAi技术敲低ITGB4。通过EdU-594、克隆形成实验探讨ITGB4对HCC增殖能力的影响;利用划痕愈合实验和Transwell等实验探讨ITGB4对HCC迁移和侵袭能力的影响。通过Western blot探讨ITGB4调控的信号通路的激活与抑制对HCC增殖、迁移等生物学效应的影响。结果 HCC中ITGB4表达升高(P<0.01),上调ITGB4激活HCC细胞中PI3K/AKT/mTOR信号通路,并促进HCC细胞的增殖和迁移(P<0.01),沉默ITGB4则显著下调PI3K/AKT/mTOR信号通路活化水平,加入PI3K/AKT/mTOR信号通路的抑制剂PKI-587后逆转了ITGB4介导的HCC增殖、迁移和侵袭等效应(P<0.01)。结论 与正常肝细胞相比,ITGB4在肝癌细胞系中高表达,并且ITGB4通过PI3K/AKT/mTOR信号通路促进HCC的发生发展。Objective To compare the expression of integrin β4(ITGB4) in hepatocellular carcinoma(HCC) and normal hepatocytes and explore the biological effects of ITGB4 in HCC and the mechanism of regulating its biological effects.Methods Three different hepatocellular carcinoma cell lines of SK-HEP-1,Huh7,HepG2 and normal liver cell L02 were selected,and the expression of ITGB4 among different cell lines was detected by Western blot.The expression of ITGB4 between Huh7 and HepG2 was detected with the immunofluorescence staining.HepG2 cells with low ITGB4 expression and Huh7 cells with high ITGB4 expression compared with normal liver cells were selected for ITGB4 overexpression and RNAi technology to knock down ITGB4 respectively.The effects of ITGB4 on the proliferation of HCC were investigated with EdU-594 in clonal formation experiments.The effects of ITGB4 on the migration and invasion of HCC were investigated with scratch healing assay and Transwell assay.The effects of activation and inhibition of ITGB4-regulated signaling pathway on the biological effects of HCC proliferation and migration were investigated with Western blot analysis.Result ITGB4 expression was increased in HCC(P<0.01),which up-regulated ITGB4 activation of PI3K/AKT/mTOR signaling pathway in HCC cells and promoted the proliferation and migration of HCC cells(P<0.01).ITGB4 silence significantly decreased the activation level of PI3K/AKT/mTOR signaling pathway.The addition of PKI-587,an inhibitor of PI3K/AKT/mTOR signaling pathway,reversed ITGB4-mediated proliferation,migration and invasion of HCC(P<0.01).Conclusions Compared with normal hepatocytes,ITGB4 is highly expressed in liver cancer cell lines promoting the development of HCC through the PI3K/AKT/mTOR signaling pathway.

关 键 词：整合素β4 肝细胞癌 PI3K/AKT/MTOR 增殖 迁移 

分 类 号：R735.7[医药卫生—肿瘤]