重组细小病毒H-1(rhH1△/p21)对胃癌细胞HGC27生长抑制作用的研究  

Inhibitory effects of recombinant human parvovirus H-1 vector expressing p21(rhH1△/ p21) on the growth of human gastric cancer cell line HGC27

作　　者：冉志华[1] 冯缨[1] 邱德凯[1] 萧树东[1] 

机构地区：[1]上海交通大学医学院附属仁济医院消化科上海市消化疾病研究所,上海200001

出　　处：《肿瘤》2007年第4期251-255,共5页Tumor

基　　金：德国大众基金会资助项目(编号:grantI/75890).;国家教育部高等学校骨干教师资助计划

摘　　要：目的:了解重组细小病毒H-1(rhH1△/p21)对胃癌细胞株HGC27的作用,为进一步研究p21wif1的生物学作用以及肿瘤的基因治疗奠定基础。方法:采用RT-PCR技术,构建表达p21蛋白的重组细小病毒H-1(rhH1△/p21),并将其转染胃癌细胞HGC27,观察细胞形态学改变,用Western blot检测转移基因蛋白在胃癌细胞中的表达,用MTT法检测其对HGC27细胞生长的抑制作用,用流式细胞仪分析其对细胞周期的影响。结果:成功构建重组细小病毒H-1(rhH1△/p21),所得病毒滴度达到3.5×107PFU/mL,在胃癌HGC27细胞中成功过表达转移基因蛋白p21,使胃癌细胞株细胞周期比例改变,G1期含量明显增加,并抑制其增殖。结论:重组细小病毒H-1(rhH1△/p21)可诱导胃癌细胞株HGC27细胞G1期阻滞,抑制细胞增殖,该基因疗法可有效抑制体外胃癌细胞的生长。。Objective：To investigate the effects of recombinant parvovirus H-1 vector expressing p21 （rhH1 △/p21 ）on human gastric cancer cell line HGC27, and to further reveal the biological function of p21^wifl to provide the basis for cancer gene therapy. Methods： The recombinant parvovirus H-lvector expressing p21 （ rhH1 △/p21 ）was constructed by reverse transcriptase-polymerase chain reaction （RT-PCR）, and was transfected into HGC27 cell line. The morphological changes of HGC27 cell were observed. The transgene protein expressions in the gastric cancer cells were detected by Western blot. The inhibitory effects of rhH1 △/p21 on the growth of HGC27 cells were measured by MTr assay. The cell cycle distribution was determined by flowcytometry. Results： The rhH1 △/p21 was successfully constructed, with a titer of 3.5 × 10^7 PFU/mL. The transgene protein p21 was over-expressed in the HC, C27 cells. The cell cycle distribution was changed. The proportion of cells in G1 phase significantly increased. The cell growth was significantly inhibited. Conclusion： rhH1△/p21 induces G1 arrest and inhibits the proliferation of gastric cancer HGC27 cells. It indicates that rhH1 △/ p21 gene therapy can effectively inhibit the growth of gastric cancer cells in vitro.

关 键 词：胃肿瘤 DNA 重组 细小病毒科感染 癌基因蛋白p21 基因表达 细胞周期 

分 类 号：R73-36[医药卫生—肿瘤]