LncRNA MIR4435-2HG在HCT-116细胞株的顺铂耐药中的作用机制

Mechanism of LncRNA MIR4435-2HG in Cisplatin Resistance of HCT-116 Cell Line

目的探讨LncRNA MIR4435-2HG在HCT-116R细胞株的顺铂耐药中的作用机制。方法通过结肠癌细胞株HCT-116,建立顺铂耐药细胞株HCT-116R,进行细胞增殖试验,细胞株分别与加或不加25μm顺铂在McCoy's 5A培养基中培养24 h,通过RNA分离和实时荧光定量PCR分析,并测定Caspase-3活性。结果顺铂耐药细胞株HCT-116R中LncRNA MIR4435-2HG水平显著增加,在耐药细胞株HCT-116R中敲除MIR4435-2HG基因可显著恢复对顺铂的敏感性,抑制细胞增殖并促进细胞凋亡。此外,mRNA水平中作为氧化应激途径的关键分子Nrf2和HO-1,被靶向MIR4435-2HG中的siRNA抑制,通过Nrf2/HO-1途径显示,MIR4435-2HG介导了顺铂耐药性。结论LncRNA MIR4435-2HG是可以驱动HCT-116的顺铂耐药性的主要因素。

Objective To investigate the mechanism of LncRNA MIR4435-2HG in cisplatin resistance of HCR116R cell line.Methods The cisplatin resistant cell line HCT-116R was established by colon cancer cell line HCT-116 to perform cell proliferation assay.The cell lines were cultured in McCoy's 5A medium with or without 25μm cisplatin for 24 hours.Separation and real-time PCR analysis were performed to determine Caspase-3 activity.Results The level of LncRNA MIR4435-2HG dramatically increased in the cisplatin resistance cell line HCT116R.Knockdown of MIR4435-2HG in HCR116R significantly restored the sensitivity to cisplatin,inhibited cell proliferation and promoted cell apoptosis.Furthermore,Nrf2 and HO-1 mRNA level,as the critical molecular of the oxidative stress pathway,was inhibited by the siRNA targeting to MIR4435-2HG,displaying MIR4435-2HG-mediated cisplatin resistance through the Nrf2/HO-1 pathway.Conclusion LncRNA MIR4435-2HG is the main factor that can drive the cisplatin resistance of HCT116.