沉默Dicer1对人卵巢癌细胞株增殖、凋亡能力及周期分布的影响

Effect of silencing Dicer1 on proliferation,apoptosis and cycle distribution of human ovarian cancer cell lines

目的探究沉默Dicer1对人卵巢癌细胞株增殖、凋亡能力及周期分布的影响。方法对购于中国医学科学院肿瘤细胞库的卵巢癌细胞进行分组,空白组卵巢癌细胞不作处理,通过细胞转染建立稳定转染的沉默组、过表达组。噻唑蓝实验(MTT)检测细胞增殖能力,流式细胞仪检测细胞凋亡、周期分布情况,Transwell小室检测细胞侵袭,细胞划痕实验检测细胞迁移,蛋白免疫印迹(Western blot)法检测B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶3(Caspase3)表达。结果在第24、48、72h,沉默组细胞增殖率均低于空白组,过表达组细胞增殖率均高于空白组、沉默组,差异均具有统计学意义(均P<0.05)。在第24、48、72h,沉默组细胞凋亡率高于空白组,过表达组细胞凋亡率均低于空白组、沉默组,差异均具有统计学意义(均P<0.05)。沉默组细胞处于G1期的比例、Bax相对表达量高于空白组,过表达组细胞处于G1期的比例、Bax相对表达量均低于空白组、沉默组,差异均具有统计学意义(均P<0.05)。沉默组细胞侵袭细胞数、迁移细胞数、Bcl-2、Caspase3相对表达量均低于空白组,过表达组细胞侵袭细胞数、迁移细胞数、Bcl-2、Caspase3相对表达量均高于空白组、沉默组,差异均具有统计学意义(均P<0.05)。结论沉默Dicer1的表达,能够抑制卵巢癌细胞增殖、侵袭和迁移,阻滞细胞周期分布,还能够调控凋亡相关蛋白Bcl-2、Bax、Caspase3的表达,促进卵巢癌细胞凋亡。

Objective To investigate the effects of silencing Dicer1 on proliferation,apoptosis and cycle distribution of human ovarian cancer cell lines.Methods Ovarian cancer cells purchased from the tumor cell bank of The Chinese Academy of Medical Sciences were grouped.The blank group of ovarian cancer cells were not treated,and the stable silence group and overexpression group were established by cell transfection.Thiazole blue experiment(determined by MTT)was used to detect cell proliferation ability,flow cytometry instrument to cell apoptosis,cycle distribution,Transwell Chambers to cell invasion,cell scratch test to cell migration,protein immunoblot(Western blot)method to the expressions of B lymphocyte tumor-2 gene(Bcl-2),the Bcl-2 X protein(Bax),cysteine,aspartic acid proteinase 3(Caspase3).Results At the 24th,48th and 72h,the cell proliferation rate of the silencing group was lower than that of the blank group,and the cell proliferation rate of the overexpression group was higher than that of the blank group and the silencing group,with statistically significant differences(all P<0.05).At the 24th,48th and 72h,the apoptosis rate of the silencing group was higher than that of the blank group,and the apoptosis rate of the overexpression group was lower than that of the blank group and the silencing group,with statistically significant differences(all P<0.05).The proportion of cells in G1 phase and the relative expression level of Bax in the silencing group were higher than those in the blank group,while the proportion of cells in G1 phase and the relative expression level of Bax in the overexpression group were lower than those in the blank group and the silencing group,with statistically significant differences(all P<0.05).The relative expressions of invasion cells,migratory cells,Bcl-2 and Caspase-3 in the silence group were all lower than those in the blank group,while the relative expressions of invasion cells,migratory cells,Bcl-2 and caspase-3 in the overexpression group were all higher than those in the blank group and the silence group,with statistically significant differences(all P<0.05).Conclusions Silencing the expression of Dicer1 can inhibit the proliferation,invasion and migration of ovarian cancer cells,block cell cycle distribution,and regulate the expression of apoptosis-related proteins Bcl-2,Bax and Caspase-3,thus promoting the apoptosis of ovarian cancer cells.