Zhang Yin, Jiang Xuefeng, Wu Zhisheng, Hu Daling, Jia Junli, Guo Jinfeng, Tang Tian, Yao Jialin, Liu Hongyi, Tang Huamin
Department of Neurosurgery, Sir Run Run Hospital, Nanjing Medical University.
School of Basic Medical Sciences, Nanjing Medical University.
J Cancer. 2020 Mar 4;11(10):2935-2944. doi: 10.7150/jca.41942. eCollection 2020.
: This study aimed to investigate whether long noncoding RNA (lncRNA) LINC00467 could regulate proliferative and invasive abilities of glioma cells via p53 and DNA methyltransferase 1 (DNMT1), so as to participate in the occurrence and progression of glioma. : LINC00467 expression in glioma was analyzed by GEPIA database and LINC00467 expression in glioma cell lines was detected by qRT-PCR. The regulatory effects of LINC00467 and p53 on proliferative, invasive capacities and cell cycle were conducted by CCK-8 and EdU assays, transwell assay and flow cytometry, respectively. The binding conditions between LINC00467, DNMT1 and p53 were determined by RNA immunoprecipitation (RIP) and Chromatin immunoprecipitation (ChIP) assays. Western blot was conducted to determine whether LINC00467 could regulate p53 in glioma cells. Finally, rescue experiments were carried out to evaluate whether LINC00467 regulates proliferative and invasive abilities of glioma cells through p53. : The expression of LINC00467 was significantly up-regulated in tumor samples than that in normal samples, which was not correlated with patient survival time. Besides, expression of LINC00467 was higher in glioma cells than that of negative control cells. Upregulation of LINC00467 promoted proliferative and invasive abilities, and accelerated cell cycle in G0/G1 phase of U87 and LN229 cells. The results of RIP and ChIP assays demonstrated that LINC00467 could bind to DNMT1 and inhibit p53 expression. Overexpression of p53 partially reversed the enhancement of LINC00467 on proliferative and invasive abilities of glioma cells. : These results indicated that high expression of LINC00467 could promote proliferative and invasive abilities of glioma cells through targeting inhibition of p53 expression by binding to DNMT1.
本研究旨在探讨长链非编码RNA（lncRNA）LINC00467是否能通过p53和DNA甲基转移酶1（DNMT1）调节胶质瘤细胞的增殖和侵袭能力，从而参与胶质瘤的发生和发展。通过GEPIA数据库分析胶质瘤中LINC00467的表达，并通过qRT-PCR检测胶质瘤细胞系中LINC00467的表达。分别通过CCK-8和EdU实验、Transwell实验及流式细胞术检测LINC00467和p53对增殖、侵袭能力及细胞周期的调节作用。通过RNA免疫沉淀（RIP）和染色质免疫沉淀（ChIP）实验确定LINC00467、DNMT1和p53之间的结合情况。采用蛋白质免疫印迹法检测LINC00467是否能调节胶质瘤细胞中的p53。最后，进行拯救实验以评估LINC00467是否通过p53调节胶质瘤细胞的增殖和侵袭能力。肿瘤样本中LINC00467的表达明显高于正常样本，且与患者生存时间无关。此外，胶质瘤细胞中LINC00467的表达高于阴性对照细胞。LINC00467的上调促进了U87和LN229细胞的增殖和侵袭能力，并加速了G0/G1期的细胞周期。RIP和ChIP实验结果表明，LINC00467可与DNMT1结合并抑制p53表达。p53的过表达部分逆转了LINC00467对胶质瘤细胞增殖和侵袭能力的增强作用。这些结果表明，LINC00467的高表达可通过与DNMT1结合靶向抑制p53表达，从而促进胶质瘤细胞的增殖和侵袭能力。
