摘要
目的:研究靶向FoxM1基因的小分子干扰RNA(siRNA)下调宫颈癌Hela、C33A及Siha等细胞系FoxM1基因表达后细胞侵袭和迁移能力的改变,寻找新的宫颈癌治疗靶点。方法:设计靶向FoxM1基因小分子干扰RNA(siFoxM1)并分别转染宫颈癌细胞系Hela、C33A和Siha,实时定量RT-PCR和Western blot技术分别检测siFoxM1对细胞内源性FoxM1表达的影响;采用Transwell小室试验检测细胞迁移和侵袭能力改变。结果:1转染siFoxM1可特异高效地下调宫颈癌细胞FoxM1mRNA水平和蛋白表达水平,Hela、C33A和Siha细胞FoxMl mRNA分别下降92.29%,87.65%和95.82%;2细胞迁移试验显示3株细胞的平均OD值较对照组分别降低40.61%,49.65%和50.87%(P<0.05)。细胞侵袭试验显示3株细胞的平均穿膜细胞数较对照组分别降低86.67%,71.77%和72.48%(P<0.01)。结论:FoxM1基因表达下调可显著降低多株宫颈癌细胞侵袭和迁移能力,有可能成为有效的宫颈癌治疗靶点。
Objective:To investigate the change of invasion ability of cervical cancer cell line Hela,C33 A,and Siha with down-regulation of FoxM1 expression.Methods:A small interfering RNA targeting FoxM1 was designed to deplete the endogenous FoxM1 expression of these cell lines.Real-time RT-PCR and Western blot were used to detect the endogenous expression of FoxM1 in the siFoxM1 cells,respectively.The migration and invasion ability of FoxM1 deficient cells were observed by transwell chamber assay.Results:The designed siRNA could dramatically deplete FoxM1 expression by 92.29%,87.65% and 95.82% in these cell lines respectively,as proved with Real-time RT-PCR and Western blot.The cell migration and invasion ability of siFoxM1 group were markedly inhibited,as manifested by transwell chamber assay.The average invasion cell numbers of siFoxM1 transfected cells decreased by 86.67%,71.77%and 72.48%(P0.01)than unrelated-siRNA transfected cells of these cell lines,respectively(P0.01).Conclusion:The invasion ability of cervical cell lines were efficiently inhibited after silencing the FoxM1 gene expression,which suggested that inhibiting FoxM1 expression might be a promising way for cer-vical cancer therapy.
出处
《武汉大学学报(医学版)》
CAS
2015年第2期254-257,262,共5页
Medical Journal of Wuhan University
基金
湖北省自然科学基金面上项目(编号:2013CFB231)