转基因细胞模型L02/HBx的构建及HBx对细胞周期的影响
HBx基因;细胞周期;细胞模型;细胞凋亡;流式细胞术;MTT法;逆转录-聚合酶链反应;免疫印迹;肝细胞癌,程斌,林松挺,杨玉珍,黎培员,程斌,通讯作者:,Establishmentofgene-transfectedcellstra
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程斌, 林松挺, 杨玉珍, 黎培员, 华中科技大学同济医学院附属同济医院消化内科 湖北省武汉市 430030
程斌, 华中科技大学同济医学院附属同济医院消化内科副教授, 主要从事乙肝相关性肝癌发生机制的基础研究及内镜超声的临床研究.
国家自然科学基金资助项目, No. 30570821
通讯作者: 程斌，430030，湖北省武汉市解放大道1095号，华中科技大学同济医学院附属同济医院消化内科.
b.cheng@tjh.tjmu.edu.cn
电话: 027-83663612 传真: 027-83646605
收稿日期: 2007-01-10 接受日期: 2007-03-17
Establishment of gene-transfected cell strain L02/HBx and effect of HBx on the cell cycles
Bin Cheng, Song-Ting Lin, Yu-Zhen Yang, Pei-Yuan Li
Bin Cheng, Song-Ting Lin, Yu-Zhen Yang, Pei-Yuan Li,Department of Gastroenterology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Supported byNational Natural Science Foundation of China, No. 30570821
Correspondence to:Dr. Bin Cheng, Department of Gastroenterology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China. b.cheng@tjh.tjmu.edu.cn
Received:2007-01-10 Accepted:2007-03-17
Abstract
AIM: To establish gene-transfected cell strain L02/HBx and study its cell cycle changes.
METHODS: Effectene transfection and G418 selection were used to obtain the positive clones of L02/HBx cells. Then HBx mRNA and protein expression were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Finally, MTT assay and flow cytometry were adopted to measure the proliferation, apoptosis and cell cycles of L02/HBx cells.
RESULTS: RT-PCR and Western blot analysis showed that the positive clones had HBx expression at mRNA and protein level. MTT assay demonstrated that the proliferation of L02/HBx cells had been accelerated. Flow cytometry found that the apoptosis rates of L02/HBx cells were at a lower level (0.09% ± 0.13% vs 3.74% ± 1.29%, P < 0.05), and the proportion of L02/HBx cells fell G1 phase (61 ......
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