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PCA3启动CD-TK自杀基因对前列腺癌细胞特异性杀伤效应初步研究
引用本文:刘雷黄星华,李坚伟陈统权李冠奕,周建华. PCA3启动CD-TK自杀基因对前列腺癌细胞特异性杀伤效应初步研究[J]. 国际泌尿系统杂志, 2018, 38(2): 177-180. DOI: 10.3760/cma.j.issn.1673-4416.2017.02.001
作者姓名:刘雷黄星华  李坚伟陈统权李冠奕  周建华
基金项目: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)
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。

关 键 词:前列腺肿瘤  基因   转基因   自杀  质粒  

Preliminary study on the specific killing effect of PCA3 promoted CD-TK suicide gene on prostate cancer cells
Abstract: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. 
Keywords:Prostatic Neoplasms  Genes   Transgenic   Suicide  Plasmids  
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