Shao Cunhua, Liu Gongpan, Zhang Xiaobin, Li Anyun, Guo Xingjun
Department of Hepatobiliary Surgery, Dongying People's Hospital, Dongying City 257091, People's Republic of China.
Department of General Surgery, Dongying Hong Gang Hospital, Dongying City 257000, People's Republic of China.
Onco Targets Ther. 2020 Apr 8;13:3013-3024. doi: 10.2147/OTT.S243736. eCollection 2020.
This study aimed to explore the regulatory effect of long noncoding RNA (lncRNA) ribonuclease mitochondrial RNA processing gene (RMRP) on hepatocellular carcinoma (HCC).
The expression of RMRP in HCC tissues and cell lines was assessed by qRT-PCR. Kaplan-Meier method was utilized to analyze the correlation between RMRP expression and the survival of HCC patients. MHCC97H and HuH7 cells were transfected with pcDNA3.1-RMRP or pcDNA3.1, respectively. MTT and flow cytometry assays were conducted to examine the proliferation and apoptosis of HCC cells, respectively. The migration and invasion of HCC cells were assessed using wound healing and transwell assays, respectively. StarBase3.0 and dual-luciferase reporter gene assay were used to identify the target relationship between miR-766 and RMRP. A xenografted tumor model was established in rats to evaluate the effect of RMRP in vivo.
RMRP was down-regulated in HCC tissues and cells. Low expression of RMRP was correlated with poor survival of HCC patients. The A495 value and colony number were significantly decreased in pcDNA3.1-RMRP-transfected MHCC97H and HuH7 cells. The apoptosis rate was significantly increased in pcDNA3.1-RMRP-transfected MHCC97H and HuH7 cells. The migration rate and the number of invasive cells were significantly decreased in pcDNA3.1-RMRP-transfected MHCC97H and HuH7 cells. MiR-766 was a target of RMRP and eliminated the anti-tumor effect of RMRP on MHCC97H cells. The up-regulation of RMRP suppressed the growth of xenograft tumors in rats.
Overexpression of RMRP suppressed the tumorigenesis of HCC by targeting miR-766.
本研究旨在探讨长链非编码RNA（lncRNA）核糖核酸酶线粒体RNA加工基因（RMRP）对肝细胞癌（HCC）的调控作用。
采用qRT-PCR检测RMRP在HCC组织和细胞系中的表达。利用Kaplan-Meier法分析RMRP表达与HCC患者生存的相关性。分别用pcDNA3.1-RMRP或pcDNA3.1转染MHCC97H和HuH7细胞。分别进行MTT和流式细胞术检测以检测HCC细胞的增殖和凋亡。分别采用伤口愈合试验和Transwell试验评估HCC细胞的迁移和侵袭能力。使用StarBase3.0和双荧光素酶报告基因试验鉴定miR-766与RMRP之间的靶向关系。在大鼠中建立异种移植肿瘤模型以评估RMRP在体内的作用。
RMRP在HCC组织和细胞中表达下调。RMRP低表达与HCC患者的不良生存相关。pcDNA3.1-RMRP转染的MHCC97H和HuH7细胞中A495值和集落数显著降低。pcDNA3.1-RMRP转染的MHCC97H和HuH7细胞凋亡率显著增加。pcDNA3.1-RMRP转染的MHCC97H和HuH7细胞迁移率和侵袭细胞数显著降低。MiR-766是RMRP的靶点，并消除了RMRP对MHCC97H细胞的抗肿瘤作用。RMRP的上调抑制了大鼠异种移植肿瘤的生长。
RMRP的过表达通过靶向miR-766抑制HCC的肿瘤发生。
