Liver Cancer
Copyright ©2008 The WJG Press and Baishideng. All rights reserved.
World J Gastroenterol. Aug 7, 2008; 14(29): 4633-4642
Published online Aug 7, 2008. doi: 10.3748/wjg.14.4633
Lentivirus mediated shRNA interference targeting MAT2B induces growth-inhibition and apoptosis in hepatocellular carcinoma
Qun Wang, Quan-Yan Liu, Zhi-Su Liu, Qun Qian, Quan Sun, Ding-Yu Pan
Qun Wang, Quan-Yan Liu, Zhi-Su Liu, Qun Qian, Quan Sun, Ding-Yu Pan, Department of General Surgery, Zhongnan Hospital, Wuhan University, Wuhan 430071, Hubei Province, China
Author contributions: Wang Q and Liu ZS contributed equally to this work; Wang Q, Liu QY and Liu ZS designed research; Wang Q, Liu QY performed research; Qian Q and Sun Q contributed new reagents/analytic tools; Wang Q, Liu QY, Liu ZS and Pan DY analyzed data; Wang Q wrote the paper.
Correspondence to: Zhi-Su Liu, Department of General Surgery, Zhongnan Hospital of Wuhan University No. 160 Dong hu Road, Wuhan 430071, Hubei Province, China. swander@126.com
Telephone: +86-27-67813007
Fax: +86-27-87330795
Received: March 13, 2008
Revised: May 27, 2008
Accepted: June 4, 2008
Published online: August 7, 2008
Abstract

AIM: To investigate the effects of lentivirus vector mediated short hairpin RNA interference targeting methionine adenosyltransferase 2β gene (LV-shMAT2B) on hepatocellular carcinoma (HCC) cells.

METHODS: We constructed four plasmids of RNA interference targeting the MAT2B gene. After LV-shMAT2B was transfected with L-02 cells and two kinds of HCC cells, cell viability and proliferation were measured with MTT and [3H]thymidine assays respectively. Flow cytometry was used to assess cell apoptosis. The level of S-adenosyl methionine (SAMe) in HepG2 cells was evaluated. The expressions of cyclin D1, cyclin D2, bcl-xL and bcl-xS were detected with western blot.

RESULTS: We constructed LV-shMAT2B successfully. LV-shMAT2B was safe for human normal liver cells. LV-shMAT2B caused dramatic reduction in proliferation compared with controls in HCC cells Bel-7402 (P = 0.054) and HepG2 (P = 0.031). Flow cytometry analysis showed that cell apoptosis caused by LV-shMAT2B was greater in HCC cells Bel-7402 and HepG2 than in control induced by scrambled siRNA (P = 0.047), but apoptosis rates in L-02 induced by LV-shMAT2B and scrambled siRNA respectively had no significant difference. Moreover, LV-shMAT2B significantly suppressed expression of MAT2B leading to growth-inhibition effect on HCC cells by down-regulating cyclin D1. Apoptosis induced by LV-shMAT2B was involved in down-regulating bcl-xL and up- regulating bcl-xS.

CONCLUSION: LV-shMAT2B can induce cell apoptosis and growth-inhibition in HCC cells. MAT2B may be a therapy target in HCC in the future.

Keywords: Lentivirus; Methionine adenosyltransferase 2β gene; Growth inhibition; Apoptosis; Hepatocellular carcinoma