长链非编码RNA RMRP和miR-34在肝细胞癌组织的表达及相关性

Expression of long non-coding RNA RMRP and miR-34 in hepatocellular carcinoma and its relationship with the prognosis

[目的]探讨长链非编码RNA(Long noncoding RNAs,Lnc RNA)线粒体RNA处理核糖核酸内切酶RNA组份(RNA component of mitochondrial RNA-processing endoribonuclease,RMRP)对肝细胞癌(HCC)细胞生物学行为的作用及与患者预后的关系。[方法]购得66例HCC组织和16例癌旁组织的cDNA芯片,采用实时荧光定量PCR(quantitative realtime PCR,qRT-PCR)法检测HCC组织中RMRP和微小RNA-34(microRNA-34,miR-34)的表达;分析RMRP表达与患者临床病理参数之间的关系;并分析RMRP表达与miR-34表达二者间的相关性。将SMMC-7721细胞系分为空白组(negative control,NC)、对照组(sh-vector)和实验组(sh-RMRP),分别为未感染慢病毒、只感染空载质粒及感染sh-RMRP-慢病毒(lentivirus,lenti),采用MTT实验和克隆形成实验检测细胞增殖能力。[结果]RMRP在HCC组织中的表达明显高于癌旁组织(t=13.41,P<0.05),且HCC组织中RMRP的表达与T分级、TNM分期及是否复发相关(P<0.05);RMRP与miR-34表达呈明显负相关(r=-0.802,P<0.05);细胞转染后培养12、24、48、72 h后,sh-RMRP组吸光度值(OD490值)明显低于NC组和sh-vector组(分别F=30.42,73.68,52.94,107.37,P<0.05);sh-RMRP组克隆形成数明显低于NC组和sh-vector组(F=257.0,P<0.05),RMRP高表达患者总体生存率明显低于RMRP低表达患者(P<0.05)。[结论]LncRNA RMRP在HCC组织中呈高表达,提示HCC患者预后不良。

[Objective]To investigate the expression of long non-coding RNA(lncRNA)RNA component of mitochondrial RNA-processing endoribonuclease(RMRP)in hepatocellular carcinoma tissues and its correlation with the prognosis of hepatocellular cancer patients.[Methods]The cDNA chip containing 66 cases of hepatocellular cancer tissues and 16 cases of para-carcinoma tissues was purchased.The expression of RMRP and miR-34 were detected by real-time fluorescence quantitative PCR(qRT-PCR),and the correlation between RMRP and clinical pathological parameters or miR-34 were analyzed.The SMMC-7721 cells were divided into blank control group(negative control,NC),control group(sh-vector)or experimental group(sh-RMRP).The cell proliferation ability was determined by MTT assay and colony formation experiment.[Results]The expression level of lncRNA RMRP in Hepatocellular carcinoma tissue was significantly higher than para-carcinoma tissues(t=13.41,P<0.05).There was a significant negative correlation between RMRP and miR-34 expression(r=-0.802,P<0.05).MTT assay and colony formation experiment demonstrated that the proliferation ability of sh-RMRP group was lower than NC group and sh-vector group(all P<0.05).The overall survival rate of patients with high RMRP expression was significantly lower than that of patients with low RMRP expression(P<0.05).[Conclusion]Hepatocellular carcinoma tissues showed higher expression of RMRP,which suggests poor prognosis of hepatocellular cancer patients.