BTg-3对肝癌细胞增殖、侵袭、迁移及Akt/GSK3β/β-catenin信号通路的影响

doi:10.11904/j.issn.1002-3070.2024.06.009

摘要/Abstract

摘要： 目的探讨B细胞转位基因3(B cell translocation gene 3,BTg-3)对肝癌细胞增殖、侵袭、迁移及蛋白激酶B/糖原合成酶激酶3β/β连环蛋白(Akt/GSK3β/β-catenin)信号通路的影响。方法 Western blot测定肝癌细胞BEL-7402、SMMC-7721和HepG2以及正常肝细胞LO2细胞中BTg-3蛋白表达水平。将HepG2细胞分为空白对照组(N组)、空载体组(V组)、BTg-3过表达组(BTg-3组)和BTg-3过表达+LICL组(BTg-3+LICL组),N组HepG2细胞不转染,V组HepG2细胞转染空载体pcDNA3.1,BTg-3组HepG2细胞转染过表达BTg-3载体pcDNA3.1-BTg-3,BTg-3过表达+LICL组转染过表达BTg-3载体pcDNA3.1-BTg-3同时给予10 nmol/LGSK-3β抑制剂氯化锂(LICL)处理。MTT法测定各组细胞增殖能力,细胞克隆形成实验测定各组细胞存活能力,Transwell小室实验测定各组细胞侵袭和迁移能力,Western blot测定各组细胞Akt、磷酸化Akt(p-Akt)、p-GSK3β、β-catenin、细胞周期素D1(Cyclin D1)、神经型钙粘蛋白(N-cadherin)和上皮型钙粘蛋白(E-cadherin)蛋白表达水平。结果人肝癌细胞BEL-7402、SMMC-7721、HepG2中BTg-3蛋白表达水平均低于正常肝细胞LO2(P＜0.001)。与N组和V组比较,BTg-3组和BTg-3+LICL组HepG2细胞增殖能力和克隆形成率降低(P＜0.05),侵袭细胞数、迁移细胞数降低(P＜0.05),p-Akt、p-GSK3β、β-catenin、Cyclin D1、N-cadherin蛋白表达水平降低(P＜0.05),BTg-3、E-cadherin蛋白表达水平升高(P＜0.05);与BTg-3组比较,BTg-3+LICL组HepG2细胞增殖能力和克隆形成率升高(P＜0.05),侵袭细胞数、迁移细胞数升高(P＜0.05),p-Akt、p-GSK3β、β-catenin、Cyclin D1、N-cadherin蛋白表达水平升高(P＜0.05),BTg-3、E-cadherin蛋白表达水平降低(P＜0.05);N组和V组HepG2细胞各指标差异无统计学意义(P＞0.05)。结论 BTg-3在肝癌细胞中低表达,过表达BTg-3可能通过Akt/GSK3β/β-catenin信号通路抑制肝癌细胞增殖、侵袭和迁移。

关键词: B细胞转位基因3, 肝癌, 上皮-间充质转化

Abstract: Objective The aim of this study was to investigate the effect of B cell translocation gene 3(BTg-3)on proliferation,invasion,migration and protein kinase B/glycogen synthase kinase-3β/β-catenin(Akt/GSK-3β/β-catenin)signaling pathway of liver cancer cells. Methods Western blot was used to measure the expression of BTg-3 protein in liver cancer BEL-7402 cells,SMMC-7721 cells,HepG2 cells and normal liver cancer LO2 cells.HepG2 cells were divided into the blank control group(N group),empty vector group(V group),BTg-3 overexpression group(BTg-3 group)and BTg-3 overexpression + LICL group(BTg-3+LICL group).HepG2 cells in the N group were not transfected and HepG2 cells of the V group were transfected with empty vector pcDNA3.1.HepG2 cells in the BTg-3 group were transfected overexpressing BTg-3 vector pcDNA3.1-BTg-3,while HepG2 cells in the BTg-3 overexpression + LiCl group were transfected with the BTg-3 vector pcDNA3.1-BTg-3 and treated with 10 nmol/l GSK-3β inhibitor-lithium chloride(LiCl).MTT assay was used to determine the proliferative ability of HepG2 cells in each group.The cell colony formation assay was used to determine the survival ability of HepG2 cells in each group.Transwell assay was used to determine the invasive and migratory ability of HepG2 cells in each group.Western blot was used to determine the expression of Akt,phosphorylated Akt(p-Akt),p-GSK-3β,β-catenin,Cyclin D1,N-cadherin and epithelial cadherin(E-cadherin)in HepG2 cells. Results The expression of BTg-3 protein in human liver cancer BEL-7402 cells,SMMC-7721 cells and HepG2 cells were lower than that in normal liver LO2 cells(P＜0.001).Compared with the N group and V group,the A values and clone formation rates were decreased in the BTg-3 group and BTg-3+LICL group(P＜0.05),as well as a decrease in the number of invasive cells and migratory cells(P＜0.05).The expression of p-Akt,p-GSK-3β,β-catenin,Cyclin D1 and N-cadherin proteins decreased(P＜0.05),and the expression of BTg-3 and E-cadherin proteins increased(P＜0.05)in the BTg-3 group and BTg-3+LICL group.Compared with the BTg-3 group,the A values and clone formation rate in the BTg-3+LICL group increased(P＜0.05),as well as an increase in the number of invasive cells and migratory cells(P＜0.05).The expression of p-Akt,p-GSK-3β,β-catenin,Cyclin D1 and N-cadherin proteins increased(P＜0.05),while the expression of BTg-3 and E-cadherin proteins in HepG2 cells decreased in the BTg-3+LICL group(P＜0.05).There were no significant differences in the indexes of HepG2 cells between the N group and V group(P＞0.05). Conclusion BTg-3 is lowly expressed in liver cancer cells,and overexpression BTg-3 may inhibit proliferation,invasion and migration of liver cancer cells through the Akt/GSK-3β/β-catenin signaling pathway.

Key words: B cell translocation gene 3, Liver cancer, Epithelial-mesenchymal transition

中图分类号:

R735.7

杨小斌, 王军平. BTg-3对肝癌细胞增殖、侵袭、迁移及Akt/GSK3β/β-catenin信号通路的影响[J]. 实用肿瘤学杂志, 2024, 38(6): 410-416.

YANG Xiaobin, WANG Junping. Effects of BTg-3 on proliferation,invasion,migration and Akt/GSK-3β/β-catenin signaling pathway of liver cancer cells[J]. Journal of Practical Oncology, 2024, 38(6): 410-416.

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