摘要
目的 研究在前体药物作用下PCA3启动子联合CD-TK基因的真核表达载体对体外前列腺癌细胞特异性杀伤效应。方法 将已构建好pHBAd-PCA3-CD-TK真核重组质粒和对照组质粒分别转染到前列腺癌细胞、对照组细胞，并以更昔洛韦（Ganciclovir，GCV）和5-氟胞嘧啶（5-Fluorocytosine，5-FC） 作为前体药物联合使用。MTT法检测并比较体外细胞毒实验后各组细胞活力。结果 成功将pHBAd-PCA3-CD-TK转染至目标细胞，采用Real-Time PCR验证其仅能在前列腺癌细胞中表达；MTT法检测可观察到pHBAd-PCA3-CD-TK组靶细胞活力明显低于对照组,差异具有统计学意义（P〈0.05）。结论 pHBAd-PCA3-CD-TK真核重组质粒可在前列腺癌细胞中特异性表达并激活前体药物，体外实验中特异性杀伤前列腺癌细胞，为后续基于PCA3启动子对前列腺癌精准基因治疗打下初步研究基础。
Objective To study the specific killing effect of the eukaryotic expression vector containing PCA3 promoter and CD-TK gene which was constructed in pre-study on prostate cancer cells with the simultaneous action of prodrug in vitro. Methods The constructed eukaryotic recombinant plasmids of pHBAd-PCA3-CD-TK and control group were transfected into prostate cancer cells and and the cells in the control group, then Ganciclovir（GCV）125 μg/mL and 5-Fluorocytosine（5-FC）160 μg/mL were delivered together as the prodrug. MTT assay was used to detect and compare the viability of cells in vitro cytotoxicity experiment.Results The pHBAd-PCA3-CD-TK was successfully transfected into target cells and verified which only expressed in prostate cancer cells by Real-Time PCR. MTT assay showed that the cell viability of pHBAd-PCA3-CD-TK group was significantly lower in control group, the difference of experimental group and control groups was statistically significant. Conclusions The pHBAd-PCA3-CD-TK can specifically express and activate prodrugs in prostate cancer cells, which can specifically kill prostate cancer cells in vitro experiments.This study lays a preliminary foundation for follow-up research on precise gene therapy of prostate cancer based on PCA3 promoter.
作者
刘雷
黄星华
李坚伟
陈统权
李冠奕
周建华
Liu Lei, Huang Xinghua, Li Jianwei, et al.(Department of Urology, People ~ Hospital of Longgang Dis- trict , Shenzhen 518172, Chin)
出处
《国际泌尿系统杂志》
2018年第2期177-180,共4页
International Journal of Urology and Nephrology
基金
广东省医学科研基金资助项目（A2015017）
深圳市科技计划项目（JCYJ20160422115256071）
龙岗区科技计划资助项目（20160607100021590）
关键词
前列腺肿瘤
基因
转基因
自杀
质粒
Prostatic Neoplasms
Genes, Transgenic, Suicide
Plasmids
