摘要
目的探讨HSVtk/GCV系统在体外诱导LoVo细胞凋亡。方法用逆转录病毒转染LoVo细胞获得阳性克隆,以GCV(10μmol·L^(-1))作用,经光镜、电镜和荧光显微镜观察各组细胞凋亡情况。HSVtk+/LoVo细胞经GCV(10μmol·L^(-1))作用0h～48h不等,荧光显微镜观察计数计算其凋亡率,并用流式细胞仪检测细胞周期。结果通过光镜、电镜和荧光显微镜观察到实验组HSVtk+/LoVo细胞凋亡情况,HSVtk+/LoVo细胞分别经GCV(10μmol·L^(-1))作用0,12,24,36,48h,凋亡率随时间的增加而升高。作用24h后凋亡率低于0.3,48h升至0.925。经流式细胞仪检测GCV作用后的肿瘤细胞,细胞周期变化明显D_0/G_1期细胞从0.31上升到0.42,S期细胞从0.62下降到0.39。结论 HSVtk/GCV系统可诱导LoVo细胞凋亡,且存在时间效应,GCV可抑制细胞DNA的合成,阻止细胞由G_1期向S期转化。
AIM To study the apoptosis of LoVo cells induced by HSVtk/GCV system in vitro. METHODS LoVo cells were transferred with retroviral vector, and followed by GCV in vitro. Apoptotic cells were observed under light microscope, electron microscopy, and fluoroscope. After HSVtk+/LoVo cells were treated with GCV for 0 to 48 hours, the percentage of apoptosis was counted with fluoroscope and cell cycle was analyzed using flow cytometry. RESULTS Apoptosis of LoVo cells were discovered under light microscope, transmission electron microscopy and fluoroscope. While HSVtk+/LoVo cells were treated by GCV for 0, 12, 24, 36 and 48 hours, the percentage of apoptosis rose gradually. In hour 24 it was less than 0.3, but rose to 0.925 in hour 48. GCV showed a remarkable cell cycle specificity. The percent of LoVo cells In G_0/G_1 phase increased from 0.31 to 0.42, and the percent of LoVo cells in S phase lowered from 0.62 to 0.39. CONCLUSION HSVtk/GCV system can induce apoptosis of LoVo cells of human colorectal carcinoma, and the percentage of apoptosis has a time effect. LoVo cells are blocked from G-1 phase to S phase.
出处
《世界华人消化杂志》
CAS
2001年第2期194-197,共4页
World Chinese Journal of Digestology
关键词
结肠直肠肿瘤
病理学
肿瘤细胞
胸苷激酶
疱疹病毒科
更西罗韦
colorectal neoplasms/pathology
tumor cells, cultured
thymidine kinase
herpesviridae
ganciclovir
apoptosis
