Ad-HSV-TK/GCV自杀基因系统对人卵巢癌细胞增殖、凋亡的影响

Experimental study of AD-HSV-TK /GCV suicide gene system on ovarian cancer cells

目的探讨腺病毒介导的单纯疱疹病毒腺苷激酶(HSV-TK)/丙氧鸟苷(GCV)自杀基因系统对人卵巢癌细胞SKOV-3增殖、凋亡的影响。方法以人卵巢癌细胞SKOV-3为实验组,人成纤维细胞株MRC-5为对照组,两组分别取对数生长期,调整细胞浓度为1×107/mL。采用Ad-hTERT-HSV-TK分别转染两组细胞48 h,然后分别给予不同浓度的GCV(0.1、1、5、10、20、50、100、200μg/mL)作用48 h。采用MTT法检测各组OD值,计算并比较两组细胞增殖率、凋亡率。结果 5、10、20、50、100、200μg/mL GCV作用下实验组细胞增殖率较对照组降低(P<0.05),实验组GCV 10、100μg/mL作用下细胞凋亡率高于对照组(P<0.05)。结论腺病毒介导HSV-TK基因转入人卵巢癌SKOV-3细胞并获稳定表达,Ad-HSV-TK/GCV自杀基因系统能明显抑制卵巢癌细胞增殖,促进卵巢癌细胞凋亡。

Objective To investigate the killing effect of adenoviral mediated herpes simplex virus adenosine kinase （HSV-TK）/ganciclovir （GCV） suicide gene system on human ovarian carcinoma cell line （SKOV-3） in vitro.Methods Human ovarian cancer cells SKOV-3 were taken as the experimental group,the human fibroblast MAC-5 as the control group,and two groups were both in logarithmic growth period,the cell concentration was 1 × 107/mL.The two groups of cells were transfected by Ad-hTERT-HSV-TK for 48 h,and then they were separately treated with different concentrations of GCV （0.1 μg,1 μg,5 μg,10 μg,20 μg,50 μg,100 μg,200 μg,500 μg and 1 000 μg/mL） for48 h.The OD value was detected by MTT,and the cell inhibitory rate and apoptosis rate between the two groups were calculated and compared.Results At different concentrations of GCV （5 μg,10 μg,20 μg,50 μg,100 μg and 200 μg/mL）,the inhibitory rate in the experimental group was lower than that of the control group （P ＜ 0.05）,the apoptosis rate was higher than that of the control group at the different concentrations of GCV （10 μg and 100 μg/mL） （P ＜ 0.05）.Conclusions The adenoviral mediated HSV-TK gene is transfected into human ovarian carcinoma cell line SKOV-3 and stably expressed.HSV-TK/GCV suicide gene system can significantly reduce the proliferation and enhance apoptosis of ovarian cancer cells.