Ge Beihai, Zhang Xian, Zhou Wei, Mo Yun, Su Z Hou, Xu Guolong, Chen Qiang
Department of Neurology, Guangxi Zhuang Autonomous Region Brain Hospital, Liuzhou, Guangxi, China.
Department of Neurosurgery, Guangxi Zhuang Autonomous Region Brain Hospital, Liuzhou, Guangxi, China.
Cell J. 2022 Jun;24(6):294-301. doi: 10.22074/cellj.2022.8035. Epub 2022 Jun 29.
This study aimed to explore biological function of long intergenic non-protein coding RNA 265 (LINC00265) in hepatocellular carcinoma (HCC) cells, and evaluate its potential function as a biomarker.
In this experimental study, GEPIA database and Kaplan-Meier Plotter database were employed to analyze LINC00265 expression in HCC tissue samples and its predicting value for prognosis. LINC00265 expression in HCC tissues and cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR). After overexpressing and knocking-down of LINC00265 in HCC cells, cell counting kit-8 (CCK-8) and 5-Ethynyl-2'- deoxyuridine (EdU) assays were adopted to detect proliferation of HCC cells. Transwell assay was used to detect migration and invasion of HCC cells. Interaction of LINC00265 with E2F transcription factor 1 (E2F1) was verified by the catRAPID online analysis tool, RNA pull-down experiment and RNA binding protein immunoprecipitation (RIP) assay. Binding of E2F1 to the promoter region of cyclin-dependent kinases 2 () was detected by dual-luciferase reporter assay and chromatin immunoprecipitation. Regulatory effects of LINC00265 and E2F1 on expression were probed by Western blot.
LINC00265 expression was increased in HCC tissues and cells. LINC00265 overexpression promoted proliferation, migration and invasion of HCC cells, and knocking-down LINC00265 worked oppositely. LINC00265 could bind to E2F1 and it could enhance combination of E2F1 and promoter regions, thus promoting CDK2 transcription. LINC00265 overexpression promoted expression of in HCC cells.
Our data suggested that LINC00265 can promote malignant behaviors of HCC cells by recruiting E2F1 to the promoter region of .
本研究旨在探讨长链基因间非编码RNA 265（LINC00265）在肝癌（HCC）细胞中的生物学功能，并评估其作为生物标志物的潜在作用。
在本实验研究中，利用GEPIA数据库和Kaplan-Meier Plotter数据库分析LINC00265在肝癌组织样本中的表达及其对预后的预测价值。采用定量实时聚合酶链反应（qRT-PCR）检测肝癌组织和细胞中LINC00265的表达。在肝癌细胞中过表达和敲低LINC00265后，采用细胞计数试剂盒-8（CCK-8）和5-乙炔基-2'-脱氧尿苷（EdU）检测法检测肝癌细胞的增殖。采用Transwell检测法检测肝癌细胞的迁移和侵袭。通过catRAPID在线分析工具、RNA下拉实验和RNA结合蛋白免疫沉淀（RIP）实验验证LINC00265与E2F转录因子1（E2F1）的相互作用。采用双荧光素酶报告基因检测法和染色质免疫沉淀检测E2F1与细胞周期蛋白依赖性激酶2（CDK2）启动子区域的结合。通过蛋白质免疫印迹法探究LINC00265和E2F1对CDK2表达的调控作用。
LINC00265在肝癌组织和细胞中的表达升高。LINC00265过表达促进肝癌细胞增殖、迁移和侵袭，敲低LINC00265则产生相反作用。LINC00265可与E2F1结合，并增强E2F1与CDK2启动子区域的结合，从而促进CDK2转录。LINC00265过表达促进肝癌细胞中CDK2的表达。
我们的数据表明，LINC00265可通过将E2F1募集至CDK2启动子区域来促进肝癌细胞的恶性行为。
