七氟醚通过调控miR-221对结直肠癌细胞增殖、凋亡的机制研究

Mechanism study of sevoflurane on proliferation and apoptosis of colorectal cancer cells by regulating miR-221

目的研究七氟醚通过调控miR-221对结直肠癌细胞增殖和凋亡的影响。方法分别采用低(1.7%)、中(3.4%)和高(5.1%)剂量的七氟醚处理HCT116细胞6 h,按要求转染后高剂量七氟醚刺激细胞。采用CCK-8、克隆形成实验和流式细胞术检测细胞增殖与凋亡。结果中、高剂量组各时间点的细胞OD值均低于低剂量组;高剂量组各时间点的细胞OD值低于中剂量组(P<0.05)。低、中、高剂量七氟醚处理组的克隆形成率低于对照组;中、高剂量组细胞克隆形成率低于低剂量组;高剂量组细胞克隆形成率低于中剂量组(P<0.05)。低、中、高剂量七氟醚处理组的细胞凋亡率高于对照组,中、高剂量组细胞凋亡率高于低剂量组,高剂量组细胞凋亡率高于中剂量组(P<0.05)。低、中、高剂量七氟醚处理组的HCT116细胞中miR-221表达水平低于对照组,中、高剂量组HCT116细胞中miR-221表达水平低于低剂量组,高剂量组HCT116细胞中miR-221表达水平低于中剂量组(P<0.05)。不同时间点间细胞OD值比较,差异有高度统计学意义(P<0.01);anti-miR-221组各时间点的细胞OD值均低于anti-miR-NC组(P<0.01)。anti-miR-221组HCT116细胞克隆形成率低于anti-miR-NC组,凋亡率高于anti-miR-NC组(P<0.01)。不同时间点间细胞OD值比较,差异有高度统计学意义(P<0.01);组间比较:miR-221组各时间点的细胞OD值高于miR-NC组(P<0.05)。miR-221组细胞克隆形成率高于miR-NC组,凋亡率低于miR-NC组(P<0.01)。不同时间点间细胞OD值比较,差异有高度统计学意义(P<0.01);组间比较,miR-221+七氟醚各时间点的细胞OD值高于miR-NC+七氟醚组(P<0.01)。miR-221+七氟醚组细胞克隆形成率高于miR-NC+七氟醚组,凋亡率低于miR-NC+七氟醚组(P<0.05)。结论七氟醚通过抑制miR-221的表达抑制结直肠癌细胞增殖,促进细胞凋亡。

Objective To investigate the effect of sevoflurane on proliferation and apoptosis of colorectal cancer cells through regulation of miR-221.Methods HCT116 cells were treated with low(1.7%),medium(3.4%)and high(5.1%)doses of sevoflurane for 6 h,respectively,and cells were stimulated with high doses of sevoflurane after transfection as required.CCK-8,clone formation assay and flow cytometry were used to detect cell proliferation and apoptosis.Results The cell OD values at all time points in the medium and high dose groups were lower than those in the low dose group;the cell OD values at all time points in the high dose group were lower than those in the medium dose group(P<0.05).The clone formation rates of the low,medium,and high dose sevoflurane-treated groups were lower than those of the control group;the cell clone formation rates of the medium and high dose groups were lower than those of the low dose group;the cell clone formation rates of the high dose group were lower than those of the medium dose group(P<0.05).The apoptosis rate in the low,medium,and high dose sevoflurane-treated groups were higher than that in the control group;the apoptosis rate in the medium and high dose groups were higher than that in the low dose group,and the apoptosis rate in the high dose group was higher than that in the medium dose group(P<0.05).The miR-221 expression levels in HCT116 cells in the low,medium,and high dose sevoflurane-treated groups were lower than those in the control group,miR-221 expression levels in HCT116 cells in the medium and high dose groups were lower than those in the low dose group,and miR-221 expression levels in HCT116 cells in the high dose group were lower than those in the medium dose group(P<0.05).The cell OD values at all time points in the anti-miR-221 group were lower than those in the anti-miR-NC group(P<0.01).The clone formation rate of HCT116 cells in the anti-miR-221 group was lower than that in the anti-miR-NC group,and the apoptosis rate was higher than that in the anti-miR-NC group(P<0.01).The cell OD values were higher in the miR-221 group than in the miR-NC group at each time point(P<0.05).The cell clone formation rate in the miR-221 group was higher than that in the miR-NC group,and the apoptosis rate was lower than that in the miR-NC group(P<0.01).The cell OD values of miR-221+sevoflurane were higher than those of miR-NC+sevoflurane group at each time point(P<0.01).The cell clone formation rate of miR-221+sevoflurane group was higher than that of miR-NC+sevoflurane group and the apoptosis rate was lower than that of miR-NC+sevoflurane group(P<0.05).Sevoflurane inhibits colorectal cancer cell proliferation and promotes apoptosis by suppressing miR-221 expression.Conclusion Sevoflurane inhibits the proliferation and promotes apoptosis of CRC cells by down-regulating miR-221.