miRNA-93调控细胞信号转导与转录激活因子3信号通路影响喉癌细胞机制研究

Study on the mechanism of miRNA-93 regulating signal transducer and activator of transcription 3 signal pathway and effects of laryngeal cancer cell proliferation

目的研究miRNA-93调控细胞信号转导与转录激活因子3(STAT3)信号通路影响喉癌细胞增殖、凋亡的作用及机制。方法选取人喉癌Hep-2细胞,分为对照组、实验组(miRNA-93组)、联合组(miRNA-93+STAT3组)。用逆转录-聚合酶链反应(RT-PCR)检测Hep-2细胞中STAT3及miRNA93的表达,蛋白质免疫印迹检测Hep-2细胞质粒-细胞信号转导与转录激活因子3(p-STAT3)、周期蛋白依赖性激酶(Cyclins)、骨髓细胞瘤病毒癌基因(c-Myc)、细胞周期蛋白依赖性激酶抑制因子(CKI)、B淋巴细胞瘤-2、生存素基因的表达。结果转染miRNA-93对STAT3基因mRNA含量无明显影响(P>0.05);转染miRNA-93可明显下调STAT3蛋白(P<0.05)。故而提示miRNA-93于转录后水平调控STAT3基因的表达;3组对Hep-2细胞周期无明显影响(P>0.05);与对照组比较,实验组可明显诱导细胞凋亡(P<0.05);而与实验组比较,联合组可明显抑制细胞凋亡(P<0.05);转染48 h后,与对照组比较,实验组转染miRNA-93后,p-STAT3、Cyclins、c-Myc、CKI、B淋巴细胞瘤-2、生存素基因明显下调,而联合组可使p-STAT3、c-Myc、B淋巴细胞瘤-2表达明显上升。结论 miRNA-93可通过与STAT3基因mRNA93结合而于转录后水平调控该基因的表达,并且通过下调STAT3信号通路的相关蛋白,从而抑制Hep-2细胞凋亡。

Objective To explore the mechanism of miRNA-93 regulating signal transducer and activator of transcription 3（ STAT3） signal pathway and effects of laryngeal cancer cell proliferation,apoptosis.Methods To select human laryngeal carcinoma Hep-2 cells,divided into control group,experimence group（ miRNA-93 group）,joint group（ miRNA-93 ＋ STAT3 group）. The mRNA level and protein level on the expression of miR-93,STAT3 and STAT3 were detected by RT-PCR and Western blot respectively. The miRNA93 Hep-2 cells were detected by reverse transcription polymerase chain reaction（ RT-PCR）.The Hep-2 expression of signal transducer and activator of transcription3,plasmid-signal transducer and activator of transcription 3,cyclin dependent kinase（ Cyclin）,bone marrow cell tumor gene（ c-Myc）,cyclin dependent kinase inhibitor（ CKI）,B lymphocyte tumor-2,survivingwere detected by Western blot. Results The transfection of miRNA-93 had no obvious change on the content of STAT3 of mRNA gene,the difference was not statistically significant（ P〉0. 05）. The transfection of miRNA-93 can downregulate STAT3 protein,the difference has statistical difference（ P〈0. 05）. It suggested that miRNA-93 expression at post transcriptional regulation of STAT3 gene. The three groups had no obvious effect on cell cycle of Hep-2（ P〉0. 05）. Compared with the control group,experience group can induce cell apoptosis,with statistically significant difference（ P〈0. 05）. Compared with experience group,joint group could significantly inhibit cell apoptosis,and the difference has statistical significance（ P〈0. 05）. After transfection of 48 h,compared with the control group,after transfection of miRNA-93,STAT3,p-STAT3,Cyclins,c-Myc,CKI,Bcl-2,survivin significantly lowered in experimence group,and in joint group can make the p-STAT3,c-Myc,Bcl-2 expression was significantly increased. Conclusion miRNA-93 combined with STAT3 gene mRNA93 in the post transcriptional regulation of the gene expression,and through the relevant protein down-regulation of STAT3 signaling pathway,thereby inhibiting the apoptosis of Hep-2 cells.