细小病毒H-1感染对胃癌细胞凋亡相关基因表达的影响  

Effects of Parvovirus H-1 Infection on the Expressions of Apoptosis-related Genes in Gastric Cancer Cells

作　　者：冯缨[1] 冉志华[1] 刘炯[1] 萧树东[1] 

机构地区：[1]上海第二医科大学附属仁济医院上海市消化疾病研究所,200001

出　　处：《胃肠病学》2005年第1期10-14,共5页Chinese Journal of Gastroenterology

基　　金：德国大众基金(grantI/75890);国家教委青年骨干教师项目资助

摘　　要：背景:细小病毒H鄄1对肿瘤细胞和转化细胞具有选择性杀伤和抑制生长的作用,是很好的基因治疗用载体。但肿瘤细胞对细小病毒H鄄1杀伤作用敏感或耐受的分子机制目前尚不十分清楚。目的:从mRNA水平观察细小病毒H鄄1感染对胃癌细胞凋亡相关基因表达的影响,探讨细小病毒H鄄1对胃癌细胞细胞毒作用的相关机制。方法:选取对细小病毒H鄄1敏感的人胃癌细胞株HGC鄄27和不敏感的人胃癌细胞株BGC鄄823,于细小病毒H鄄1感染48h后提取细胞总RNA。应用不同荧光染料标记的dUTP逆转录制备荧光探针,与含有8000点人类体细胞基因序列的基因表达谱芯片杂交,再经计算机荧光扫描以分析敏感细胞株HGC鄄27凋亡相关基因表达谱的改变。采用逆转录聚合酶链反应(RT鄄PCR)对敏感细胞株HGC鄄27部分凋亡相关基因,如SARP1、BCL鄄10、CL鄄20、NCDRP和RAIDD等的表达作进一步检证,并比较其与不敏感细胞株BGC鄄823相应凋亡相关基因的表达差异。结果:基因表达谱芯片检测结果显示,HGC鄄27细胞感染细小病毒H鄄148h后,所检测的64对凋亡相关基因中有15对基因表达水平下调至原水平的50%以下,仅有1对基因表达水平上调至2倍以上。RT鄄PCR扩增结果显示,感染细小病毒H鄄148h后,敏感细胞株HGC鄄27的SARP1、BCL鄄10、CL鄄20和NCDRP基因表达明显下调。Parvovirus H-1 can selectively kill or inhibit the growth of tumor and transformed cells, and may serve as an ideal vector for gene therapy, but the molecular mechanism of the response of tumor cells to parvovirus H-1 infection (sensitive or insensitive) remains unclear. Aims: To observe the effects of parvovirus H-1 infection on the expression of apoptosis-related gene in gastric cancer cells at mRNA level, and to explore the mechanism of parvovirus H-1 induced cytotoxic effects in infected gastric cancer cells. Methods: Parvovirus H-1 -sensitive human gastric cancer cell line HGC-27 and insensitive human gastric cancer cell line BGC-823 were employed in this study. The cellular RNA was extracted 48 hours after virus inoculation. The differentially labeled fluorescent dUTP reverse transcription generated probes were used to hybridize with cDNA microarray containing 8 000 spots of human somatic gene sequences. The fluorescence of genechip was scanned by a specialized computer, the changes of apoptosis-related genes expression profiles were analyzed in the sensitive cell line HGC-27. The expressions of some apoptosis-related genes in the sensitive cell line HGC-27, such as SARP1, BCL-10, CL-20, NCDRP and RAIDD were further verified by reverse transcriptase polymerase chain reaction (RT-PCR), and compared with those in the insensitive cell line BGC-823. Results: The results of cDNA microarray indicated that 15 out of 64 apoptosis-related genes were down-regulated (less then 0.5 fold), whilst only one gene was up-regulated (more then 2 folds) in sensitive cell line HGC-27 48 hours after parvovirus H-1 infection. The results of RT-PCR ampli-fication showed that the expressions of SARP1, BCL-10, CL-20 and NCDRP genes were markedly decreased, whilst the expression of RAIDD gene had no apparent change in H-1 virus infected sensitive cell line HGC-27; however, in the insensitive cell line BGC-823, the expressions of BCL-10 and NCDRP genes were increased at the same time point, the expressions of CL-20 and RAIDD genes h

关 键 词：胃肿瘤 细小病毒H-1 细胞凋亡 基因表达 逆转录聚合酶链反应 

分 类 号：R735.2[医药卫生—肿瘤]