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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 investigate the killing effect of HSV TK / GCV system on bladder cancer cell line BIU 87 and its effect on apoptosis gene. Methods The gene of TK was successfully transferred into BIU 87 and the effects of GCV on apoptosis and cell cycle were detected by MTT assay, TUNEL assay and flow cytometry (FCM). The expression of reverse transcriptase polymerase The changes of bcl 2, bax, bak and Caspase 3 mRNA were detected by reverse transcription polymerase chain reaction (RT PCR). The changes of Caspase 3 activity were measured by colorimetric assay. RESULTS: After treated with 1 mg / LGCV for 5 days, 67% of cells were killed and 100% of LGCV killed 97% of cells. GCV induced apoptosis and cell cycle arrest in S and G2 phases. The expression of bcl- bax expression increased, Caspase 3 expression and activity increased. Conclusion HSV TK / GCV system can effectively kill bladder cancer cells and cause apoptosis, cell cycle arrest, bcl 2 and bax expression changes, and Caspase 3 activation is an important factor in apoptosis.