摘要
目的探讨爪蟾驱动样蛋白2靶向蛋白(Targeting protein for Xenopus kinesin-like protein 2,TPX2)在肝细胞癌中的表达及作用,并初步探讨其促进肝癌进展的分子机制。方法应用实时定量RT-PCR法及Western blot法检测肝细胞系L02、肝癌细胞系HepG2、MHCC-97H、Huh7、SMMC-7721中TPX2的表达情况;应用TPX2-siRNA转染MHCC-97H细胞,通过Western-blot技术检测转染后TPX2的表达变化,并采用BrdU细胞增殖分析、Caspase3/7活性分析检测细胞增殖及凋亡变化,明确下调TPX2表达对肝癌细胞增殖凋亡的影响;应用c-MYC-siRNA转染MHCC-97H细胞,通过Western-blot技术分析c-MYC的表达与TPX2分子及AKT信号通路的关系。结果肝癌细胞系MHCC-97H中TPX2表达水平显著高于其它细胞系L02、HepG2、MHCC-97H、Huh7、SMMC-7721(P<0.05);下调TPX2表达将显著降低MHCC-97H细胞的增殖,并促进肝癌细胞凋亡;下调c-MYC表达能够显著下调TPX2表达并减少AKT的磷酸化水平,抑制AKT信号通路活化。结论TPX2可能在肝细胞癌的增殖凋亡中起了重要作用,很有可能是肝癌靶向治疗的一个新的靶点。
Objective To investigate the expression of TPX2 and its effect in HCC,explore the role of TPX2 in HCC development and analysis the possible mechanisms underlying this process.Methods The expression of TPX2 was detected by quantitative RT-PCR and western blot in liver cell line L02,liver cancer cell lines HepG2,MHCC-97H,Huh7,SMMC-7721.TPX2-siRNAs were transfected into MHCC-97H cells.The expression of TPX2 were identified by Western blot,cell proliferation and apoptosis were analyzed by BrdU incorporation assay and Caspase 3/7 activity assay.C-MYC-siRNAs were transfected into MHCC-97H cells.Western blot was used to investigate the mechanism of TPX2 in regulating the AKT Signaling Pathway and its upstream proteins c-MYC.Results The cell line MHCC-97H showed significantly higher TPX2 expression level than the cell lines L02,HepG2,MHCC-97H,Huh7 and SMMC-7721(P<0.05).Downregulation of TPX2 repressed MHCC-97H cell proliferation and facilitated cell apoptosis.Knockdown of c-MYC down-regulated the expression of TPX2 and inhibited the phosphorylation of AKT.Conclusion TPX2 may play a key role in the HCC’s proliferation and apoptosis and may be considered as a new target for the treatment of HCC.
作者
刘清泉
张旭峰
梁嘉毅
谢斌辉
何晓
LIU Qingquan;ZHANG Xufeng;LIANG Jiayi(Department of Hepatobiliary Surgery,The First Affiliated Hospital,of Gannan Medical University,Ganzhou,Jiangxi 341000,China)
出处
《江西医药》
CAS
2021年第9期1313-1316,1320,共5页
Jiangxi Medical Journal
基金
江西省卫生健康委科技计划项目,编号:20185343
江西省自然科学基金青年项目,编号:20192BAB215030
国家自然科学基金项目,编号:82060505。
