摘要
目的探讨利用慢病毒介导小干扰核糖核酸(siRNA)沉默人体端粒酶反转录酶(h TERT)的可行性及其对宫颈癌Caski细胞增殖和凋亡的影响。方法用前期构建的慢病毒表达载体anti-h TERT-LV和NC-LV转染宫颈癌Caski细胞。实验共分3组,分别为空白对照组(NC组)、阴性对照组(NC-LV组)及干扰组(anti-h TERT-LV组)。采用实时荧光定量PCR检测h TERT mRNA表达量,采用流式细胞仪检测细胞周期及细胞凋亡情况。结果 anti-h TERT-LV组h TERT mRNA相对表达量为(0.28±0.02)明显低于NC组(1.0)及NC-LV组(0.87±0.05)(P均<0.05)。anti-h TERT-LV组G1期细胞比例(71.52±0.79)%明显高于NC组(54.69±2.84)%和NC-LV组(55.70±1.31)%(P均<0.05);anti-h TERT-LV组S期细胞比例(18.69±1.95)%明显低于NC组(36.06±1.56)%和NC-LV组(34.16±1.09)%(P均<0.05)。anti-h TERT-LV组细胞凋亡率(36.45±5.39)%明显高于NC-LV组(8.75±3.75)%和NC组(9.55±2.57)%(P均<0.05)。结论慢病毒介导siRNA可以沉默h TERT基因,高效特异地抑制宫颈癌Caski细胞h TERT基因的表达,促进细胞凋亡,抑制肿瘤细胞的生长和增殖。
Objective To explore the feasibility of lentivirus-mediated small interfering RNA（ siRNA） silencing the human telomerase reverse transcriptase（hTERT） and its influence on the proliferation and apoptosis of cervical cancer Caski cells.Methods The cervical cancer Caski cells were transfected by lentivirus expression vectors anti-hTERT-LV and NC-LV which had been constructed in advance.There were three groups in the experiment,blank group（NC group）,negative control group（NC-LV group） and interference group（anti-hTERT-LV group）.The expression level of hTERT mRNA was determined by real-time fluorescent quantitative polymerase chain reaction,while the cell cycle and apoptosis were detected by flow cytometry respectively.Results The relative expression level of hTERT mRNA in the anti-hTERT-LV group was significantly lower than those in the NC group and NC-LV group（0.28 ±0.02,1.0,0.87 ±0.05,all P〈0.05）.The proportion of the G1-stage cells in the anti-hTERT-LV group was significantly higher than those in the NC group and NC-LV group（（71.52 ±0.79）%, （54.69 ±2.84）%,（55.70 ±1.31）%,all P〈0.05）.The proportion of the S-stage cells in the anti-hTERT-LV group was significantly lower than those in the NC group and NC-LV group（（18.69 ±1.95）%,（36.06 ±1.56）%,（34.16 ±1.09）%,all P〈0.05）.The rate of cell apoptosis in the anti-hTERT-LV group was significantly higher than those in the NC group and NC-LV group（（36.45 ±5.39）%, （8.75 ±3.75）%,（9.55 ±2.57）%,all P〈0.05）.Conclusion The lentivirus-mediated siRNA can induce silencing hTERT gene, effectively suppress the expression of hTERT gene in cervical cancer Caski cells,induce the cell apoptosis,and inhibit the growth and proliferation of the tumor cells.
出处
《广西医学》
CAS
2015年第5期599-602,共4页
Guangxi Medical Journal
基金
广西自然科学基金(2013GXNSFAAO19254)
