Li Ziwei, Xu Yong, Wang Qin, Xie Changli, Liu Yincui, Tu Zhiguang
Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Pingshan People's Hospital, Guangdong 518118, China.
Saudi J Biol Sci. 2017 Jan;24(1):95-102. doi: 10.1016/j.sjbs.2016.09.003. Epub 2016 Sep 21.
To investigate the effect of over-expression of tissue factor pathway inhibitor-2 (TFPI-2) on the differentiation of hepatocellular carcinoma (HCC) cells (Hep3B and HepG2). The TFPI-2 recombinant adenovirus (pAd-TFPI-2) was constructed using the pAdeasy-1 vector system. Transfected by pAd-TFPI-2, the cell proliferation of HCC cells was evaluated by CCK-8 assay, flow cytometry was used to detect cell apoptosis and CD133 expression. Real-time PCR and Western blot were used to detect the expression levels of markers of hepatocellular cancer stem cells (CSC) and hepatocytes. The over-expression of TFPI-2 significantly suppressed cell proliferation, induced apoptosis, and dramatically decreased the percentage of CD133 cells, which was considered as CSC in HCC. Real-time PCR and Western blot showed that the expression of markers of CSC in Hep3B cells and HepG2 cells infected with pAd-TFPI-2 was markedly lower than those of the control group ( < 0.05), while the expression of markers of hepatocytes was significantly increased ( < 0.05). Hence, TFPI-2 could induce the differentiation of hepatocellular carcinoma cells into hepatocytes, and is expected to serve as a novel way for the treatment of HCC.
探讨组织因子途径抑制物-2（TFPI-2）过表达对肝癌细胞（Hep3B和HepG2）分化的影响。采用pAdeasy-1载体系统构建TFPI-2重组腺病毒（pAd-TFPI-2）。用pAd-TFPI-2转染后，通过CCK-8法评估肝癌细胞的增殖，采用流式细胞术检测细胞凋亡和CD133表达。采用实时荧光定量PCR和蛋白质免疫印迹法检测肝癌干细胞（CSC）和肝细胞标志物的表达水平。TFPI-2过表达显著抑制细胞增殖，诱导细胞凋亡，并显著降低CD133阳性细胞（被认为是肝癌中的CSC）的比例。实时荧光定量PCR和蛋白质免疫印迹法显示，pAd-TFPI-2感染的Hep3B细胞和HepG2细胞中CSC标志物的表达明显低于对照组（<0.05），而肝细胞标志物的表达显著增加（<0.05）。因此，TFPI-2可诱导肝癌细胞向肝细胞分化，有望成为治疗肝癌的新途径。
