Expression Silence of DNA Repair Gene hMGMT Induced by RNA Interference

• Published in: Chinese journal of cancer research Vol. 19; no. 1; pp. 52 - 55
• Main Author: 李秀英 赖延东
• Format: Journal Article
• Language: English
• Published: Institute for Genome Medicine Research, School of Pharmaceutical, Jinan University, Guangzhou 510632%Institute for Chemical Carcinogenesis, Guangzhou Medical College, Guangzhou 510182 01.03.2007
• Subjects: DNA修复基因; hMGMT; RNA干扰; 表达沉默; 16HBE; RNA interference; O6-methylguanine-DNA methyl transferase (MGMT); siRNA expression cassettes
• ISSN: 1000-9604; 1993-0631
• DOI: 10.1007/s11670-007-0052-2

Objective： MGMT protein expression has been associated with tumor resistance to alkylating agents. The objective of this paper is to construct the RNA interference vector which can specifically induce the expression silence of human DNA repair gene hMGMT. Methods： The hMGMT specific siRNA expression cassette was made by two steps PCR, linked with pUCI 9 to get pU6-MGMTi, co-transfected with pEGFP-CI into 16HBE and screened by G418. The MGMT mRNA and protein levels were detected by RT-PCR and Western Blot respectively. Results： hMGMT specific RNA interfere vector pU6-MGMTi was constructed successfully. In transfected 16HBE cells MGMT mRNA level could hardly be detected and the protein level was only 10% of control. Conclusion： MGMT specific RNAi expression cassette can effectively inhibit MGMT expression. MGMT silence cell line was built by co-transfection technology, which offered condition for studying the gene function of MGMT.