RNA干扰Livin基因联合顺铂对喉鳞状细胞癌Hep-2细胞增殖和凋亡的影响

Influence of RNA interference targeting Livin combined with cisplatin treatment on proliferation and apoptosis of Hep-2 cells in laryngeal squamous cell carcinoma

目的探讨RNA干扰Livin基因联合顺铂对喉鳞状细胞癌细胞系Hep-2细胞增殖和凋亡的影响。方法取人喉鳞状细胞癌Hep-2细胞株1株进行体外培养,取对数生长期细胞随机分为对照组(细胞未经任何处理)、干扰组(细胞经RNA干扰Livin转染)、顺铂组(细胞经6μmol/L顺铂处理)、联合组(细胞经RNA干扰Livin转染+6μmol/L顺铂处理),每组设置6个复孔。处理48 h后,采用四甲基偶氮唑盐微量酶反应比色法(MTT法)检测各组细胞增殖抑制率,采用流式细胞仪双染法检测细胞凋亡率和细胞周期分布,采用逆转录聚合酶链反应(RT-PCR)检测细胞Bcl-2 mRNA、Bax mRNA表达。结果联合组细胞增殖抑制率、细胞凋亡率均高于干扰组、顺铂组,差异有统计学意义(P<0.05)。联合组G;/G;期细胞占比均高于干扰组、顺铂组,S期和G;/M期细胞占比均低于干扰组、顺铂组,差异有统计学意义(P<0.05)。联合组Bcl-2 mRNA水平低于干扰组、顺铂组,Bax mRNA水平高于干扰组、顺铂组,差异有统计学意义(P<0.05)。结论RNA干扰Livin基因联合顺铂处理喉鳞状细胞癌Hep-2细胞,可抑制细胞增殖并促进细胞凋亡,其机制可能与调节凋亡相关基因以及改变细胞分裂周期有关。

Objective To investigate the influence of RNA interference targeting Livin combined with cisplatin on proliferation and apoptosis of Hep-2 cells in laryngeal squamous cell carcinoma.Methods One Hep-2 cell line of laryngeal squamous cell carcinoma was cultured in vitro,and cells in the logarithmic phase were collected and randomly divided into control group(without any treatment),interference group(transfected by RNA interference Livin),cisplatin group(treated with6μmol/L of cisplatin),and combined group(transfected with RNA interference Livin and 6μmol/L of cisplatin),with 6 replicate holes for each group.After 48 hours of treatment,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was used to detect the inhibition rate of cell proliferation.Flow cytometry double staining was used to detect the apoptosis rate and cell cycle distribution.Reverse transcriptase polymerase chain reaction(RT-PCR)was used to detect the expression of Bcl-2 mRNA and Bax mRNA.Results The cell proliferation inhibition rate and apoptosis rate were higher in the combined group than those in the interference group and the cisplatin group(P<0.05).The percentage of cells in G;/G;phase was higher,and the percentages of cells in S phase and G;/M phase were lower in the combined group than those in the interference group and the cisplatin group(P<0.05).The Bcl-2 mRNA level was lower and the Bax mRNA level was higher in the combined group than those in the interference group and the cisplatin group(P<0.05).Conclusion The interference targeting Livin combined with cisplatin for Hep-2 cells in laryngeal squamous cell carcinoma can inhibit cell proliferation and promote cell apoptosis.The mechanism may be related to the regulation of apoptosis-related genes and the alteration of cell division cycle.