Maxizyme对肝癌突变抑癌基因p53的抑制作用
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李 岩, 林菊生, 孔心涓
Maxizyme;突变型p53;肝细胞癌,李岩,林菊生,孔心涓,通讯作者,:,Inhibitoryeffectofmaxizymeonmutan
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李岩, 林菊生, 孔心涓, 华中科技大学同济医学院附属同济医院肝病研究所 湖北省武汉市 430030
李岩, 女, 1981-12-25生, 河南省驻马店人, 汉族, 华中科技大学同济医学院硕士生, 研究方向分子肝脏病学.
国家自然科学基金资助项目, No. 30171061
通讯作者: 林菊生, 430030, 湖北省武汉市, 华中科技大学同济医学院附属同济医院肝病研究所. linjusheng2001@163.com
电话: 027-83662578
收稿日期: 2005-04-04 接受日期: 2005-04-09
Inhibitory effect of maxizyme on mutant-type p53 in hepatocellular carcinoma
Yan Li, Ju-Sheng Lin, Xin-Juan Kong
Yan Li,Ju-Sheng Lin, Xin-Juan Kong, Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Supported by National Natural Science Foundation of China, No.30171061
Correspondence to: Ju-Sheng Lin, Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province,China. linjusheng2001@163.com
Received: 2005-04-04 Accepted: 2005-04-09
Abstract
AIM: To investigate the inhibitory effect of maxizyme on the mutant-type p53 (mtp53) gene at codon 249 in exon 7 (AGG→AGT) in cell-free system, hepatocellular carcinoma (HCC) cell line MHCC97, and nude mice bearing human HCC, and to explore a new method for gene therapy of HCC.
METHODS: Anti-mtp53 and control mutant maxizyme were designed and then cloned into the vector pBSKU6 and pEGFPC1, respectively. The 32p-labeled mtp53 transcript was the target mRNA. Cold maxizyme transcript was incubated with 32p-labeled target RNA in cell-free system. The products were quantified by measuring the radioautographed count per minute (cpm) in 1 mL solution. The MHCC97 cells were the target cells which contained a mutation at the third-base position of codon 249 of the p53 gene(AGG→AGT). PEGFPMz (recombinant eukaryotic vector) was transfected into MHCC97 cells by LipofectamineTM2000. The expression of mtp53 was analyzed by Northern Blot and Western Blot ......
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