Expression Silence of DNA Repair Gene hMGMT Induced by RNA Interference
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...
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| Published in | Chinese journal of cancer research Vol. 19; no. 1; pp. 52 - 55 |
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| Main Author | |
| 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
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1000-9604 1993-0631 |
| DOI | 10.1007/s11670-007-0052-2 |
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| Abstract | 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. |
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| AbstractList | 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. R730.1; 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 pUC19 to get pU6-MGMTi, co-transfected with pEGFP-C1 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. |
| Author | 李秀英 赖延东 |
| AuthorAffiliation | Institute for Genome Medicine Research, School of Pharmaceutical, Jinan University, Guangzhou 510632 Institute for Chemical Carcinogenesis, Guangzhou Medical College, Guangzhou 510182 |
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| Notes | Q756 RNA interference siRNA expression cassettes 11-2591/R 16HBE O^6-methylguanine-DNA methyl transferase （MGMT） RNA interference; O^6-methylguanine-DNA methyl transferase （MGMT）; 16HBE; siRNA expression cassettes ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
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| Publisher | Institute for Genome Medicine Research, School of Pharmaceutical, Jinan University, Guangzhou 510632%Institute for Chemical Carcinogenesis, Guangzhou Medical College, Guangzhou 510182 |
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| SubjectTerms | DNA修复基因 hMGMT RNA干扰 表达沉默 |
| Title | Expression Silence of DNA Repair Gene hMGMT Induced by RNA Interference |
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