胸苷激酶基因系统对膀胱癌细胞及凋亡基因的影响  

HSV-TK-mediated changes of apoptosis gene and cell killing effect on BIU87 cells

作　　者：毛晓鹏[1] 丘少鹏[1] 曹开源[2] 袁广卿[2] 陈显景[1] 徐林[2] 

机构地区：[1]中山大学附属第一医院泌尿外科,广州510080 [2]中山大学医学院检验系

出　　处：《中华实验外科杂志》2005年第2期180-182,i002,共4页Chinese Journal of Experimental Surgery

基　　金：广东省自然科学基金资助项目 (0 1 0 71 9)

摘　　要：目的　探讨单纯疱疹病毒胸苷激酶 /丙氧鸟苷 (HSV TK/GCV )系统对膀胱癌细胞BIU 87的杀伤作用及对凋亡基因的影响。方法　脂质体将TK基因成功转入BIU 87,噻唑蓝(MTT)法、TUNEL法、流式细胞仪 (FCM )分别检测GCV对其生长影响及凋亡率和细胞周期变化 ;逆转录 聚合酶链反应 (RT PCR)检测bcl 2、bax、bak、Caspase 3mRNA表达变化 ,比色法测定Cas pase 3活性变化。结果　作用 5d 1mg/LGCV能杀死 67%的细胞 ,10 0mg/LGCV达到最大杀伤率杀死 97%的细胞 ;GCV诱导细胞凋亡 ,细胞周期阻滞在S和G2期 ;bcl 2表达降低 ,bax表达升高 ,Caspase 3表达及活性升高。结论　HSV TK/GCV系统可有效杀伤膀胱癌细胞并引起细胞凋亡 ,细胞周期阻滞、bcl 2和bax表达变化、Caspase 3活化是引起凋亡发生的重要因素。Objective To study the killing effect andmechanism of HSV-TK/GCV suicide gene therapy on BIU87 cells.Methods TK gene was successfully transfected into BIU87 cells with LIPOFECTAMINE 2000.The cytotoxicity and bystander effect were estimated by MTT.Apoptosis was detected by using in situ cell apoptosis detection kit,and confirmed by flow cytometry.bcl-2,bax,bak,Caspase-3,bak mRNA changes were detected by RT-PCR,and Caspase-3 activity was estimated by chromometry.Results After 5 days,67 % cells was killed by 1 mg/L GCV and 97 % by 100 mg/L GCV.Apoptosis,S- and G2 -phase arrest were induced after GCV treatment.There were apparent changes in bcl-2,bax,Caspase-3 mRNA expression and an increase of Caspase-3 activity.Conclusion HSV-TK/GCV system can inhibit the proliferation of BIU87 cells by inducing apoptosis,which may be correlated with S-and G2-phase arrest and changes of bcl-2,bax,finally induce apoptosis by activating Caspase-3.

关 键 词：膀胱癌细胞 GCV 凋亡基因 Caspase-3 BAX表达 胸苷激酶基因 升高 细胞周期阻滞 逆转录一聚合酶链反应 细胞凋亡 

分 类 号：R737.14[医药卫生—肿瘤] R735[医药卫生—临床医学]