下调ANKRD22表达对人肺腺癌细胞增殖、凋亡及细胞周期的影响

Effects of down-regulating ANKRD22 expression on proliferation,apoptosis,and cell cycle of human lung adenocarcinoma cells

目的通过RNA干扰技术下调人非小细胞肺癌(NSCLC)细胞株H1975中锚蛋白重复系列22(ANKRD22)表达,观察其对细胞增殖、细胞凋亡及细胞周期的影响。方法体外培养H1975细胞,随机分为siRNA干扰组、阴性对照组、空白对照组,siRNA干扰组、阴性对照组分别转染ANKRD22-siRNA质粒和阴性对照质粒NC-siRNA,转染24 h;空白对照组不予转染。采用实时荧光定量qRT-PCR法检测各组ANKRD22表达,采用MTT法检测细胞培养24、48、72、96、120 h的细胞增殖情况,流式细胞术检测细胞凋亡率和细胞周期。结果 siRNA干扰组ANKRD22相对表达量低于阴性对照组和空白对照组(P均<0. 05),空白对照组与阴性对照组比较P> 0. 05。siRNA干扰组转染24 h再培养24、48、72、96、120 h细胞增殖能力均低于阴性对照组和空白对照组(P均<0. 05),空白对照组与阴性对照组比较P均> 0. 05。siRNA干扰组G0/G1期细胞所占比例高于空白对照组与阴性对照组(P均<0. 05),S期、G2/M期细胞所占比例低于空白对照组与阴性对照组(P均<0. 05),空白对照组与阴性对照组比较P均> 0. 05。siRNA干扰组细胞凋亡率高于空白对照组与阴性对照组(P均<0. 05),空白对照组与阴性对照组比较P> 0. 05。结论通过RNA干扰下调NSCLC细胞株H1975中ANKRD22表达后,可抑制细胞增殖,促进细胞凋亡,延长细胞周期。

Objective To investigate the effects of down-regulating the expression of ankyrin repeat domain 22(ANKRD22)on the proliferation,apoptosis,and cell cycle of human non-small-cell lung cancer(NSCLC)H1975 cells.Methods H1975 cells were randomly divided into the the siRNA interference group,negative control group,and blank control group.We transfected the ANKRD22-siRNA and NC-siRNA into the siRNA interference group and negative control group by Lipofectamine 2000 reagent;the blank control group was not transfected.The expression level of ANKRD22 was detected by qRT-PCR.MTT assay was used to detect the proliferation of cells cultured for 24,48,72,96 and 120 h.Flow cytometry was used to detect the apoptotic rate and cell cycle.Results The expression level of ANKRD22 in the siRNA interference group was significantly lower than that in the blank control group and negative control group(both P<0.05),and no significant difference was found between the blank control group and negative control growp,P >0.05;the cell proliferation rate in the siRNA interference group was significantly lower than that in the blank control group and negative control group(P<0.05);the proportion of cells in the G 0/G 1 phase of the siRNA interference group was significantly higher than that in the blank control group and negative control group,and the proportion of cells in S phase and G 2/M phase was significantly lower than that in the blank control group and negative control group;the apoptosis rate of the siRNA interference group was significantly higher than that of the blank control group and the negative control group(P<0.05);no significant differences were found in the above parameters between the blank control group and negative control group(all P >0.05).Conclusion The down-regulation of ANKRD22 expression can inhibit the proliferation of H1975 cells significantly,promote the apoptosis,and prolong the cell cycle.