SiRNA沉默一切顺势盒2基因表达对宫颈癌细胞增殖与迁移侵袭和凋亡的影响

Effects of ONECUT2 siRNA on proliferation, migration, invasion and apoptosis in cervical cancer cells

目的研究siRNA技术沉默一切顺势盒2(ONECUT2)对宫颈癌HeLa细胞增殖、迁移、侵袭和凋亡的影响。方法将宫颈癌HeLa细胞分为4组:si-NC组和低、中、高3个浓度siRNA ONECUT2组。si-NC组细胞转染si-NC阴性对照,siRNA ONECUT2组用低、中、高3个浓度(15,30,45 nmol·L-1)siRNA ONECUT2转染48 h。用实时定量聚合酶链反应法测定ONECUT2基因表达水平;用蛋白质印迹法检测蛋白表达水平,用噻唑蓝法、Transwell实验和流式细胞术分别检测HeLa细胞增殖、迁移、侵袭和凋亡水平。结果 si-NC组和低、中、高3个浓度siRNA ONECUT2组的ONECUT2基因表达量分别为1.00±0.06,0.42±0.08,0.28±0.04和0.33±0.05,其中的中浓度siRNA ONECUT2组效果最好。si-NC组和中浓度siRNA ONECUT2组细胞在72 h的增殖抑制率分别为(16.54±0.31)%和(38.56±0.41)%;这2组的迁移细胞数量分别为245.69±13.46和103.56±8.36;这2组的侵袭细胞数分别为162.86±11.34和67.89±8.56;这2组细胞凋亡率分别为(5.89±0.61)%和(32.86±2.34)%;这2组的循环蛋白D1表达量分别为1.00±0.07和0.37±0.09;这2组的MMP-2蛋白表达量分别为1.00±0.06和0.34±0.07;这2组的Caspase-3蛋白表达量分别为1.00±0.07和2.13±0.13。上述指标:siRNA ONECUT2组与si-NC组相比,差异均有统计学意义(均P<0.05)。结论 siRNA沉默ONECUT2基因可抑制HeLa细胞的增殖、迁移和侵袭并促进细胞凋亡。

Objective To investigate the effect of silencing one cut homeobox 2(ONECUT2) by siRNA on the proliferation, invasion and apoptosis in cervical carcinoma HeLa cells.Methods Cervical cancer HeLa cells were divided into four groups: si-NC group and low,medium and high concentration siRNA ONECUT2 groups.The si-NC group was transfected with si-NC.The siRNA ONECUT2 groups were transfected with siRNA ONECUT2 at low, medium and high concentrations(15,30,45 nmol·L-1) for 48 h.The expression of ONECUT2 mRNA was detected by quantitative real-time PCR.The expression of proteins were detected by Western blotting.The proliferation, migration, invasion and apoptosis of HeLa cells were detected by MTT,Transwell and flow cytometry,respectively.Results The expressions of ONECUT2 mRNA in si-NC group and low, medium and high concentrations siRNA ONECUT2 groups were 1.00±0.06,0.42±0.08,0.28±0.04 and 0.33±0.05,respectively;the inhibition effect of siRNA ONECUT2 group in medium concentration was the best.The proliferation inhibition rates of siRNA ONECUT2 in si-NC group and medium siRNA ONECUT2 group at 72 h were(16.54±0.31) % and(38.56±0.41) %,respectively;the number of migrating cells in the two groups was 245.69±13.46 and 103.56±8.36,respectively;the number of invasive cells in the two groups was 162.86±11.34 and 67.89±8.56,respectively;the apoptosis rates in the two groups were(5.89±0.61) % and(32.86±2.34) %,respectively;the protein expressions of Cyclin D1 in the two groups were 1.00±0.07 and 0.37±0.09;the protein expressions of MMP-2 in the two groups were1.00±0.06 and 0.34±0.07;the protein expressions of Caspase-3 in the two groups were 1.00±0.07 and2.13±0.13,respectively.Comparing between siRNA ONECUT2 group and si-NC group,the differences of the factors were statistically significant(all P < 0.05).Conclusion siRNA silencing ONECUT2 inhibits the proliferation,migration and invasion of HeLa cells and promotes apoptosis.