沉默Dyrk1B基因对宫颈癌细胞生物学功能的影响

Effect of silencing Dyrk1B on biological function of cervical cancer cells

目的通过沉默在宫颈癌C4-1和Hela细胞Dyrk1B基因,观察Dyrk1B基因对宫颈癌细胞生物学功能的影响。方法应用慢病毒转染宫颈癌C4-1和Hela细胞沉默Dyrk1B基因作为沉默组,同时以空载病毒转染组为对照组。通过荧光显微镜观察2组转染效率,采用RT-PCR法检测Dyrk1B mRNA表达水平,采用流式细胞仪检测细胞凋亡率,四甲基偶氮唑蓝(MTT)比色法检测细胞增殖能力以及对DDP的敏感性。结果慢病毒转染宫颈癌C4-1和Hela细胞转染效率无差异,沉默组Dyrk1B mRNA表达均低于空载对照组,宫颈癌C4-1和Hela细胞沉默组细胞的凋亡率较空载对照组增加,而宫颈癌C4-1细胞沉默组增殖较空载对照组降低,宫颈癌C4-1和Hela细胞沉默组较空载对照组比较IC50值均降低。结论慢病毒转染宫颈癌C4-1和Hela细胞沉默Dyrk1B基因表达可增加凋亡率,而细胞增殖能力下降,并且提高宫颈癌C4-1和Hela细胞对顺铂的敏感性。

Objective To observe the effect of silencing Dyrk1B gene on the biological function of cervical cancer C4-1 and Hela cells. Methods Dyrk1B gene of human cervical cancer C4-1 and Hela cells silenced by Lentivirus carriing were enrolled into the silence group. The control group was transfected with an empty lentivirus. The transfection efficiency of two groups was observed by fluorescence microscopy. Real Time fluorescent quantitative PCR(qPCR) was used to detect mRNA expression of Dyrk1B gene, and flow cytometry was used to detect cell apoptosis rate, while methyl thiazolyl tetrazolium(MTT) colorimetry was used to detect their sensitivity to DDP. Results There was no difference in the transfection efficiency of C4-1 and Hela cells transfected by lentiviral. The expression of Dyrk1B mRNA in the silence group was lower than that in the control group. The apoptosis rate of C4-1 and Hela cells in silence group was higher than that in the control group, but the proliferation of the C4-1 cell in silence group was lower than that of the control group. The IC50 values of the C4-1 and Hela cells in the silence group were lower than those in the control group. Conclusion The silencing of Dyrk1B gene by lentiviral transfection increases the apoptosis rate and decreases the cell proliferation ability of cervical cancer C4-1 and Hela cells. And silenceing Dyrk1B gene can increase the sensitivity of C4-1 and Hela cells to cisplatin.