pre-hsa-miR-98对卵巢癌细胞增殖活性抑制的影响

Inhibitory effect of pre-hsa-miR-98 transfection on the proliferation of ovarian cancer cell lines

目的卵巢癌的死亡率占各类妇科恶性肿瘤的首位,对女性患者的生命造成严重威胁。miR-98可抑制多种癌细胞增殖,但未见关于其应用于卵巢癌治疗的报道。文中为探讨卵巢癌治疗的新方法,用pre-hsa-miR-98转染卵巢癌SKOV3、HO8910和A2780细胞,观察转染率并研究转染后对卵巢癌细胞的增殖抑制作用。方法利用构建的pre-hsa-miR-98质粒HmiR和连接随机序列的质粒CmiR转染人卵巢癌细胞SKOV3、HO8910和A2780细胞,荧光显微镜与流式细胞仪计数计算转染率,MTT法检测细胞增殖抑制率与细胞计数绘制生长曲线研究转染对卵巢癌细胞的增殖抑制作用。结果转染HmiR后,荧光显微镜观察和流式细胞术计数结果显示SKOV3、HO8910、A2780细胞的转染率均在60%左右。转染HmiR后,MTT法检测SKOV3、HO8910、A2780细胞的增殖抑制率随转染时间延长而增加,72 h分别达到49.54%、53.25%、46.09%。生长曲线显示在72 h及96 h抑制率达到高峰,随后随时间延长抑制率不再显著升高。结论 3种卵巢癌细胞的转染率达到实验要求,HmiR转染后可抑制卵巢癌细胞增殖,其抑制作用随转染时间延长而增强。

Objective Ovarian cancer ranks first in mortality among gynecological malignancies,and poses a vital threat to the lives of the patients.Though miR-98 can inhibit the proliferation of various cancer cell lines,its effect on ovarian cancer has never been reported.This study is to search for a new treatment of ovarian cancer by transfecting pre-hsa-miR-98 into ovarian cancer SKOV3,HO8910 and A2780 cells and observing the transfection efficiency and its inhibitory effect on the proliferation of ovarian cancer cells.Methods The pre-hsa-miR-98 plasmid HmiR and plasmid CmiR from a control vector were transfected into human ovarian cancer cell lines SKOV3,HO8910 and A2780.The transfection efficiency was determined by fluorescence microscopy and flow cytometry,and the inhibitory effect on the proliferation of the cell lines detected by MTT assay and the cell count growth curve.Results After HmiR transfection,the rates of transfection in SKOV3,HO8910 and A2780 cell lines were all about 60%,and the rates of the cell proliferation inhibition rose with the increased time of transfection,up to 49.54%,53.25%,and 46.09% after 72 hours of transfection.The cell growth inhibition reached the peak at 72 and 96 hours after the treatment,and thereafter exhibited no significant rise with the increase of time.Conclusion The rates of transfection of the three ovarian cancer cell lines by pre-hsa-miR-98 HmiR were all up to the experimental requirements,and the transfection inhibited the proliferation of ovarian cancer cell lines in a time-dependent manner.