长链非编码RNA XIST通过充当miR-497-5p的分子海绵并靶向程序性细胞死亡蛋白4（PDCD4）来调节肝癌细胞的增殖和迁移。
lncRNA XIST regulates proliferation and migration of hepatocellular carcinoma cells by acting as miR-497-5p molecular sponge and targeting PDCD4.
作者信息
Zhang Yixi, Zhu Zebin, Huang Shanzhou, Zhao Qiang, Huang Changjun, Tang Yunhua, Sun Chengjun, Zhang Zhiheng, Wang Linhe, Chen Huadi, Chen Maogen, Ju Weiqiang, He Xiaoshun
机构信息
1Organ Transplant Center, The First Affiliated Hospital, Sun Yat-sen University, No. 58 Zhongshan Er Road, Guangzhou, 510080 China.
2Guangdong Provincial Key Laboratory of Organ Donation and Transplant Immunology, Guangzhou, China.
出版信息
Cancer Cell Int. 2019 Jul 29;19:198. doi: 10.1186/s12935-019-0909-8. eCollection 2019.
BACKGROUND
MicroRNAs (miRNAs) play a pivotal role in hepatocellular carcinoma (HCC) progression and have been confirmed to participate in the carcinogenesis and development of HCC. However, the relationship between miR-497-5p and HCC remains unclear.
METHODS
Kaplan-Meier curve analysis and the log-rank test were used to investigate the efficacy of miR-497-5p on overall survival (OS) and disease-free survival (DFS) in patients with HCC. According to in vitro experiments, programmed cell death 4 (PDCD4) was a target of miR-497-5p by the dual-luciferase activity assay. The efficacy of PDCD4 on cell proliferation and metastasis in HCC was examined by transwell assays, CCK-8 assays and reverse transcription quantitative PCR (RT-qPCR). Additionally, we conducted a luciferase activity reporter assay to confirm the interaction between lncRNA XIST and miR-49-5p. Then, to evaluate the relationship between lncRNA XIST and miR-497-5p, several mechanistic experiments, including qRT-PCR, Western blotting, transwell assays and tumor xenograft assays, were performed.
RESULTS
miR-497-5p was upregulated in HCC tissues, and high expression of miR-497-5p resulted in increases in tumor size and tumor number and a higher tumor-node-metastasis (TNM) stage and Edmondson grade in patients with HCC. Silencing miR-497-5p inhibited the proliferation and migration of HCC cells. PDCD4, which was downregulated in HCC tissues, was shown to be a target of miR-497-5p and was negatively correlated with the expression of miR-497-5p. lncRNA XIST was found to act as a miR-497-5p sponge and to regulate the level of PDCD4, which is targeted by miR-497-5p. lncRNA XIST was observed to be downregulated in the HCC tissues and positively correlated with the expression of PDCD4.
CONCLUSIONS
Our findings reveal that the XIST/miR-497-5p/PDCD4 axis participates in HCC development and that XIST could be used as a biomarker of HCC.
背景
微小RNA（miRNA）在肝细胞癌（HCC）进展中起关键作用，并且已被证实参与HCC的发生和发展。然而，miR-497-5p与HCC之间的关系仍不清楚。
方法
采用Kaplan-Meier曲线分析和对数秩检验来研究miR-497-5p对HCC患者总生存期（OS）和无病生存期（DFS）的影响。根据体外实验，通过双荧光素酶活性测定法确定程序性细胞死亡4（PDCD4）是miR-497-5p的靶标。通过Transwell实验、CCK-8实验和逆转录定量聚合酶链反应（RT-qPCR）检测PDCD4对HCC细胞增殖和转移的作用。此外，我们进行了荧光素酶活性报告基因实验以证实长链非编码RNA XIST与miR-49-5p之间的相互作用。然后，为了评估长链非编码RNA XIST与miR-497-5p之间的关系，进行了包括qRT-PCR、蛋白质免疫印迹法、Transwell实验和肿瘤异种移植实验在内的多项机制实验。
结果
miR-497-5p在HCC组织中上调，miR-497-5p的高表达导致HCC患者肿瘤大小和肿瘤数量增加，肿瘤-淋巴结-转移（TNM）分期和Edmondson分级更高。沉默miR-497-5p可抑制HCC细胞的增殖和迁移。在HCC组织中表达下调的PDCD4被证明是miR-497-5p的靶标，并且与miR-497-5p的表达呈负相关。发现长链非编码RNA XIST作为miR-497-5p的海绵发挥作用，并调节miR-497-5p靶向的PDCD4的水平。观察到长链非编码RNA XIST在HCC组织中表达下调，并且与PDCD4的表达呈正相关。
结论
我们的研究结果表明，XIST/miR-497-5p/PDCD4轴参与HCC的发展，并且XIST可作为HCC的生物标志物。
Zotero 插件