目的:检测肝癌细胞中转录因子KLF6的表达水平,确定KLF6对肿瘤转移相关基因Basigin-2的转录调控机制,并探讨其对Basigin-2介导的肝癌细胞侵袭转移的调控作用。方法:培养肝癌细胞及正常肝细胞,提取总RNA,real-timeRT-PCR方法确定KLF6和Basigin-2的表达水平。荧光素酶报告基因系统检测KLF6对Basigin-2的转录调控作用。过表达或干涉KLF6基因,观察Basigin-2表达水平的变化,并用Transwell方法检测KLF6对肝癌细胞体外侵袭力的影响。结果:相对于正常肝细胞,肝癌细胞中Basigin-2在mRNA水平表达上调,而KLF6表达水平下调。KLF6可以与Basigin-2启动子区结合,抑制Basigin-2的转录活性。肝癌细胞中过表达KLF6可以在mRNA水平上抑制Basigin-2的转录表达,而干涉KLF6则表现为Basigin-2的表达上调。过表达KLF6可以抑制肝癌细胞的体外侵袭力,相反干涉KLF6促进了肝癌细胞的侵袭转移。结论:转录因子KLF6抑制Basigin-2的转录激活,KLF6可以抑制Basigin-2介导的肝癌细胞的侵袭转移。
Objective:To explore the expression of transcription factor KLF6,determine the regulation of KLF6 on tumor metastasis related gene Basigin - 2 and observe roles of KLF6 on hepatocellular carcinoma cells invasion and metastasis. Methods:Hepatocellular carcinoma cells and normal hepatocytes were cultured. In purified total RNA,ex-pression levels of KLF6 and Basigin - 2 were detected using real - time RT - PCR. The transcriptional regulation of KLF6 on Basigin - 2 were inspected with luciferase reporter assay system. Through overexpressing or interfering KLF6,the expression regulation of KLF6 on Basigin - 2 were observed,and the role of KLF6 on hepatocellular carci-noma cells invasion was explored using in vitro transwell invasion assay. Results:Compared to normal hepatocyte con-trol,the expression levels of Basigin - 2 were obviously up - regulated in mRNA level,however those of KLF6 were down - regulated. Transcription factor KLF6 could bind to genome DNA located on the promoter region of Basigin - 2, and repress Basigin - 2 transcription. Overexpressing KLF6 in HCC cells could inhibit the expression of Basigin - 2, while knocking down KLF6 could up - regulate the expression of Basigin - 2. Overexpressed KLF6 also could inhibit hepatocellular carcinoma cells invasion and metastasis. Conclusion:Transcription factor KLF6 repressed the transcrip-tional regulation of Basigin - 2. KLF6 could inhibit Basigin - 2 mediated hepatocellular carcinoma cells invasion and metastasis.