Knockdown of HBx by RNAi inhibits proliferation and enhances chemotherapy-induced apoptosis in hepatocellular carcinoma cells.
The present study was designed to research on RNA interference hepatitis B virus x gene approach to hepatocellular carcinoma (HCC) therapy. Previously, we constructed and identified shRNA eukaryotic expression vectors (pshRNA-X220) specific to HBx gene, pshRNA-MOCK (control); and established HCC cell lines with stable expression shRNA eukaryotic vector targeting HBx gene-21543 cell lines (MHCC97-H of expressing shRNA against HBx), HK3 cell lines (MHCC97-H by transfected with pshRNA-MOCK). We examined the expression of HBx gene after RNA interference by semi-quantitative RT-PCR and assessed the effect of HBx knocked down on cell growth by proliferation assay using kit-8 (CCK8). As well as, we analyzed cell cycle distribution by flowcytometry and examined cell apoptosis using TUNEL assay. The HBx mRNA expression level is reduced, and cells growth was significantly stopped in 21543 cell lines. Cells with HBx knockdown were more sensitive to 5-fluorouracil/cisplatin. RNA interfering HBx induced an obvious time and dose-dependent inhibitory in comparison with the control cells. Meanwhile, RNA interferenced targeting HBx, in combination with chemotherapy can effectively induce apoptosis in hepatocellular carcinoma cells and restricts cell proliferation.
He
Yan
Y
The Liver Disease Research Institute, Second Xiangya Hospital, Central South University, No 139 Renmin Road, 410011, Changsha, Hunan Province, China. hy3753@yahoo.com.cn
Sun
Hui-qing
HQ
He
Xing-e
XE
Wang
Wen-long
WL
Lei
Jian-hua
JH
D000964:Antimetabolites, Antineoplastic / Q000627:therapeutic use; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D017209:Apoptosis; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology* / Q000628:therapy; D002453:Cell Cycle; D049109:Cell Proliferation; D002945:Cisplatin / Q000627:therapeutic use; D005260:Female; D005434:Flow Cytometry; D005472:Fluorouracil / Q000627:therapeutic use; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000628:therapy; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D054334:Viral Regulatory and Accessory Proteins
D000964:Antimetabolites, Antineoplastic; D000970:Antineoplastic Agents; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; D002945:Cisplatin; D005472:Fluorouracil