负载人细胞色素P4502B6基因的复制缺陷型腺病毒感染前列腺癌PC-3细胞的研究  

Effects of replication-deficient adenovirus carrying cytochrome P450 gene on human prostate cancer cell line PC-3

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作  者:卞军[1] 刘存东[1] 杨建昆[1] 周其赵[1] 薛康颐[1] 郭文彬[1] 

机构地区:[1]南方医科大学第三附属医院泌尿外科,广州510630

出  处:《中华实验外科杂志》2013年第10期2146-2147,共2页Chinese Journal of Experimental Surgery

基  金:广东省医学科研基金资助项目(A2010360)

摘  要:目的 探讨负载人细胞色素P450(CYP2B6)基因的复制缺陷型腺病毒感染PC-3前列腺癌细胞的效果及CYP2B6表达.方法 将负载CYP2B6基因的复制缺陷型腺病毒以200 VP/cell的感染复数(MOI),感染人前列腺癌PC-3细胞;采用实时荧光定量聚合酶链反应(Real-time PCR)检测PC-3细胞内CYP2B6的表达丰度,并以Western blot检测CYP2B6蛋白表达水平.结果 与阴性对照腺病毒比较,负载CYP2B6基因的腺病毒感染PC-3细胞后,可显著提高PC-3细胞CYP2B6mRNA表达水平(8.12±2.41比0.92±0.19,P<0.05)及CYP2 B6蛋白表达水平(0.82±0.23比0.12±0.06,P<0.05);而未感染腺病毒的空白对照组与阴性对照组PC-3细胞表达CYP2B6 mRNA(0.78±0.27比0.92±0.19)和CYP2B6蛋白(0.14±0.03比0.12 ±0.06)的水平比较差异均无统计学意义(P>0.05).结论 以复制缺陷型腺病毒为载体,可向前列腺癌PC-3细胞中有效导入CYP2B6基因,并在PC-3细胞中表达.Objective To indentify effects of the replication-deficient recombinant adenovirus carrying cytochrome P450 (CYP2B6) gene on human prostate cancer cell line PC-3.Methods The human prostate cancer cell line PC-3 cells were infected with the replication-deficient recombinant adenovirus carrying CYP2B6 gene by the MOI of 200 VP/cell.The expression level of CYP2B6 mRNA and protein was detected by using real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting respectively.Results Compared to the negative control,after infection of the replication-deficient recombinant adenovirus carrying CYP2B6 gene,PC-3 cells expressed higher level of CYP2B6 mRNA (8.12 ± 2.41 vs.0.92 ±0.19,P <0.05) and CYP2B6 protein (0.82 ±0.23 vs.0.12 ±0.06,P <0.05).No significant difference was observed between the ZERO group (PC-3 cells not infected with adenovirus) and negative control in CYP2B6 mRNA expression level (0.78 ±0.27 vs.0.92 ±0.19,P >0.05) and protein expression level (0.14 ± 0.03 vs.0.12 ± 0.06,P > 0.05).Conclusion The replication-deficient recombinant adenovirus carrying CYP2B6 gene can infect human prostate cancer cell line PC-3 effectively,and these PC-3 cells can express CYP2B6 mRNA and CYP2B6 protein effectively.

关 键 词:前列腺癌 细胞色素P450基因 复制缺陷型腺病毒 

分 类 号:R737.25[医药卫生—肿瘤]

 

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