Pre-let-7d转染卵巢癌细胞转染率的研究

目的研究Pre-let-7 d质粒在不同时间,以不同浓度转染人卵巢癌细胞的转染率。方法将构建的Pre-let-7 d质粒HmiR和连接随机序列的质粒CmiR转染人卵巢癌SKOV3、HO8910细胞,用荧光显微镜与流式细胞仪计数两种方法计算转染率。结果转染HmiR、CmiR后,荧光显微镜观察和流式细胞术计数结果显示SKOV3、HO8910细胞的转染率,以3μg质粒/孔转染SKOV3、HO8910人卵巢癌细胞比2μg、4μg质粒/孔的转染率高,24 h、48 h、72 h三个不同时间点用相同质量的质粒转染,转染率无显著差异。结论转染HmiR、CmiR后,荧光显微镜观察和流式细胞术计数结果显示以3μg质粒/孔转染SK-OV3 HO8910人卵巢癌细胞的转染率均在60%左右,符合实验要求。

Experimental investigation on the transfection rates of pre-let-7 d in human ovarian cancer cells

Objective To investigate the pre-let-7d plasmid transfection rate in human ovarian cancer cells for the sake of preparing standards of transfaction dosages and hours to be adopted in the future researches;Methods The human ovarian cancer cell lines SKOV3and HO8910 cells were transfected by pre-let-7d plasmid HmiR and the random sequence plasmid CmiR.The transfection rates was detected with fluorescence microscope and FCM.Results The fluorescence microscope and flow cytometry count results show that,the transfection rate in HO8910 and SKOV3,with 3 μg plasmid/hole is higher compared to 2 μg and 4 μg plasmid/hole.There were not significant differences with the same quality transfection at 24 h,48 h and 72 h.Conclusion The transfection rate with 3 μg plasmid/hole in SKOV3 and HO8910 with HmiR、CmiR were both reach to 60%,which meet the requirements of the experiments.