转录共激活子TAZ对肝癌细胞株MHCC97H增殖能力的影响

摘要

目的 观察转录共激活子TAZ对肝癌细胞株MHCC97H的增殖能力的影响,以期对阐明原发性肝细胞癌(HCC)的发生发展机制提供1个新的研究思路.方法 应用细胞转染、RNA干扰(RNAi)以及过表达技术,在肝癌细胞株MHCC97H中干预TAZ的表达.在转染后提取细胞RNA和总蛋白,应用qRT-PCR和Western blot等方法检测TAZ、Nek7的表达.选用细胞计数kit-8(CCK-8)法检测细胞活性和增殖能力.结果 与NC组相比,转染了TAZ特异性靶向干扰片段之后,TAZ的mRNA与蛋白表达水平均出现下调,差异具有统计学意义(P＜0.0001);Nek7表达也出现下降,差异具有统计学意义(P＜0.0001);功能实验(CKK8)发现MHCC97H细胞活性和增殖能力也降低,差异具有统计学意义(P＜0.001).另一方面,过表达了TAZ之后,Nek7表达水平也相应上调了,差异具有统计学意义(P＜0.0001);MHCC97H细胞活性和增殖能力也提高,差异具有统计学意义(P＜0.001).结论 转录共激活子TAZ能通过调控Nek7的表达对肝癌细胞MHCC97H的增殖能力产生影响,该发现对于阐明TAZ在肝癌发生发展过程中的作用提供一定的实验基础.%Objective To study the effect of transcriptional co-activator TAZ(transcriptional coactivator with PDZ binding motif)on the proliferation of hepatocellular carcinoma cell line MHCC97H,to provide a new research idea for clarifying the occurrence and development mechanism of hepatocellular carcinoma(HCC).Methods Change the expression of TAZ in hepatocellular carcinoma cell line MHCC97H by using cell transfection,RNA interference(RNAi)and over expression.After transfection,the RNA and total proteins were extracted,and then the expression of TAZ and Nek7 were detected by qRT-PCR and western blot.Cell counting kit-8(CCK-8)assay was performed to detect the cell viability and proliferation.Results Compared with the non-control(NC)group,the TAZ's mRNA and protein expression were down regulated after transfected the TAZ specific targeting interfere fragments,the difference was statistically significant(P＜0.0001)correspondingly,the expression of Nek7 were decreased;meanwhile the functional tests(CKK8)showed that the viability and proliferation of MHCC97H cells were also decreased.On the other hand,the expression of Nek7 was increased after up-regulating the TAZ's expression;as a result,the viability and proliferation ability of MHCC97H cells was also increased.Conclusion The transcriptional co-activator TAZ can affect the proliferation of HCC cell line MHCC97H by regulating the expression of Nek7,which can provide some experimental basis for elucidating the role of TAZ in the progression of hepatocellular carcinoma.