Niu Yuexiang, Tang Gongen, Wu Xiuli, Wu Chaoyu
Department of Infectious Diseases, Linyi Central Hospital, Linyi, Shandong, P.R. China.
Department of Respiration, Yishui people's Hospital, Linyi, Shandong, P.R. China.
Saudi J Gastroenterol. 2020 May 26;26(4):194-203. doi: 10.4103/sjg.SJG_4_20.
BACKGROUND/AIMS: The purpose of this study is to explore the expression characteristics of lncRNA NEAT1 in hepatocellular carcinoma (HCC) and the molecular mechanism of its regulation on sorafenib resistance.
This experimental study was performed from June 2013 to June 2019. The level of NEAT1 was determined using RT-PCR in HCC and matched adjacent tissues from 79 HCC patients in Linyi central hospital. The patients were divided into two groups to compare their prognosis based on the median NEAT1 expressions as a cutoff value. HCC cell line HepG2 negative control (HepG2-NC), sorafenib-resistant HepG2 cells (HepG2-SR) were transfected with or without NEAT1 siRNA, followed by subsequent molecular analysis, to determine the function of NEAT1 on sorafenib resistance in HCC cells. The cell transcripts were determined by RNA-sequencing analysis. The binding site of the NEAT1 and microRNA-149-5p (miR-149-5p) was verified by luciferase assay.
We found that NEAT1 was significantly increased in HCC tissues. Furthermore, NEAT1 expressions were significantly associated with HCC prognosis and chemoresistance patterns against sorafenib. Subsequently, the sorafenib-resistant HCC cell lines, together with the controls, were used to determine the regulatory effect of NEAT1 on HCC cells' progression and sorafenib resistance. NEAT1 targets the miR-149-5p, and therefore, decrease the activity of sorafenib against HCC cells. NEAT1 functions were demonstrated to be triggered by the regulation of miR-149-5p/AKT1 axis.
NEAT1/miR-149-5p/AKT1 pathway-based therapy might be a potential clinical application for HCC patients.
背景/目的：本研究旨在探讨长链非编码RNA NEAT1在肝细胞癌（HCC）中的表达特征及其对索拉非尼耐药的调控分子机制。
本实验研究于2013年6月至2019年6月进行。采用逆转录聚合酶链反应（RT-PCR）检测了临沂市中心医院79例HCC患者的HCC组织及配对癌旁组织中NEAT1的水平。以NEAT1表达中位数为界值将患者分为两组比较预后。对HCC细胞系HepG2阴性对照（HepG2-NC）、索拉非尼耐药的HepG2细胞（HepG2-SR）转染或不转染NEAT1小干扰RNA（siRNA），随后进行分子分析，以确定NEAT1对HCC细胞索拉非尼耐药的作用。通过RNA测序分析确定细胞转录本。通过荧光素酶报告基因实验验证NEAT1与微小RNA-149-5p（miR-149-5p）的结合位点。
我们发现NEAT1在HCC组织中显著升高。此外，NEAT1表达与HCC预后及对索拉非尼的化疗耐药模式显著相关。随后，利用索拉非尼耐药的HCC细胞系及对照来确定NEAT1对HCC细胞进展及索拉非尼耐药的调控作用。NEAT1靶向miR-149-5p，从而降低索拉非尼对HCC细胞的活性。NEAT1的功能被证明是由miR-149-5p/蛋白激酶B1（AKT1）轴的调控触发的。
基于NEAT1/miR-149-5p/AKT1通路的治疗可能是HCC患者潜在的临床应用。
