摘要
【目的】探讨下调长链非编码HOX转录反义RNA(HOX transcript antisense RNA,HOTAIR)表达对人卵巢癌SKOV3/DDP细胞顺铂耐药的逆转作用及可能的机制。【方法】针对长链非编码RNA HOTAIR基因序列体外合成小干扰RNA(small interfering RNA,siRNA),脂质体法将siRNA转染人卵巢癌SKOV3/DDP细胞。荧光定量PCR方法检测HOTAIR表达,Western blotting检测ERK1/2和p-ERK1/2蛋白表达。MTT法检测SKOV3/DDP细胞的顺铂半数致死量(IC_(50)),应用流式细胞仪检测SKOV3/DDP细胞凋亡率。【结果】人卵巢癌SKOV3/DDP细胞经转染HOTAIR siRNA后,HOTAIR表达降低(P<0.05),同时SKOV3/DDP细胞ERK1/2蛋白和p-ERK1/2蛋白表达也明显降低(P<0.05)。与对照组比较,siRNA干扰组SKOV3/DDP细胞对顺铂的IC_(50)值下降(P<0.05),细胞凋亡率增加(P<0.05)。【结论】下调长链非编码RNA HOTAIR表达能够逆转人卵巢癌SKOV3/DDP细胞对顺铂的耐药,抑制ERK1/2途径可能是其作用机制。
【Objective】To explore the effect of down-expression of long noncoding RNA（lnc RNA） HOTAIR on reversing cisplatin resistance in human ovarian carcinoma SKOV3/DDP cells and its possible mechanism.【Methods】The siRNA targeting lnc RNA HOTAIR was synthetized in vitro and transfected into human ovarian carcinoma SKOV3/DDP cells by Lipofectamine 2000.The real time PCR was used to detect HOTAIR expression,and Western blotting was used to detect the expressions of ERK1/2 protein and pERK1/2 protein.Furthermore,the cisplatin half cell lethal dose（IC_（50）） was assessed by MTT,and the apoptotic rate was detected by flow cytometry.【Results】 After the SKOV3/DDP cells were transfected with siRNA targeting HOTAIR,the expression of HOTAIR decreased（P0.05）,and the expressions of ERK1/2 protein and p-ERK1/2 protein also decreased（P0.05）.Meanwhile,compared with the control group,the IC_（50） of SKOV3/DDP cells to cisplatin decreased（P0.05）,and the apoptotic rates increased in the siRNA group（P0.05）.【Conclusion】Down-regulation of lnc RNA HOTAIR can reverse the cisplatin resistance in human ovarian carcinoma SKOV3/DDP cells,and its potential mechanism may b e related with the inhibition of ERK1/2 pathway.
出处
《武警后勤学院学报（医学版）》
CAS
2017年第10期845-848,共4页
Journal of Logistics University of PAP（Medical Sciences）
