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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
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）
