Li Lin, Wan Ke, Xiong Linkai, Liang Shuang, Tou Fangfang, Guo Shanxian
Department of Thoracic Oncology, Jiangxi Cancer Hospital, Nanchang 330029, People's Republic of China.
Onco Targets Ther. 2020 Apr 3;13:2873-2886. doi: 10.2147/OTT.S243533. eCollection 2020.
Circular RNAs (circRNAs) have been found to regulate several human tumors. The present study was to explore the mechanism of hsa_circ_0087862 in regulating non-small cell lung cancer (NSCLC).
Totally 102 NSCLC cases were enrolled. NCI-H1359 and A549 cells were transfected. Cells viability, apoptosis, migration and invasion were determined by CCK-8 assay, flow cytometry, scratch test and transwell experiment, respectively. Luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay were performed. Xenograft tumor experiments were performed using nude mice. hsa_circ_0087862, miR-1253 and RAB3D expression in tissues/cells were detected by qRT-PCR. RAB3D and Ki67 protein expressions in cells/tissues were researched by Western blot and immunohistochemistry. Apoptosis of xenograft tumor tissue cells was detected using Tunel assay.
hsa_circ_0087862 was significantly up-regulated in NSCLC patients, which was associated with poor prognosis ( < 0.05). hsa_circ_0087862 down-regulation prominently weakened NSCLC cells viability, migration, invasion and enhanced apoptosis ( < 0.01). hsa_circ_0087862 overexpression exhibited the opposite results in NSCLC cells. miR-1253 was sponged by hsa_circ_0087862. miR-1253 expression in NSCLC tissues was negatively correlated with hsa_circ_0087862 ( < 0.001). RAB3D expression in NSCLC was directly inhibited by miR-1253. miR-1253 down-regulation or RAB3D overexpression dramatically reversed NSCLC cells phenotype induced by hsa_circ_0087862 down-regulation. hsa_circ_0087862 down-regulation markedly inhibited tumor growth in vivo ( < 0.01). In xenograft tumor tissues, hsa_circ_0087862 down-regulation obviously decreased expression of RAB3D, Ki67 and increased apoptosis.
hsa_circ_0087862 acted as an oncogene in NSCLC by targeting miR-1253/RAB3D.
环状RNA（circRNAs）已被发现可调节多种人类肿瘤。本研究旨在探讨hsa_circ_0087862在调节非小细胞肺癌（NSCLC）中的机制。
共纳入102例NSCLC病例。对NCI-H1359和A549细胞进行转染。分别通过CCK-8法、流式细胞术、划痕试验和Transwell实验检测细胞活力、凋亡、迁移和侵袭能力。进行荧光素酶报告基因检测和RNA免疫沉淀（RIP）检测。使用裸鼠进行异种移植瘤实验。通过qRT-PCR检测组织/细胞中hsa_circ_0087862、miR-1253和RAB3D的表达。通过蛋白质印迹法和免疫组织化学研究细胞/组织中RAB3D和Ki67蛋白的表达。使用Tunel法检测异种移植瘤组织细胞的凋亡情况。
hsa_circ_0087862在NSCLC患者中显著上调，这与预后不良相关（<0.05）。hsa_circ_0087862下调显著削弱NSCLC细胞活力、迁移、侵袭能力并增强凋亡（<0.01）。hsa_circ_0087862过表达在NSCLC细胞中表现出相反的结果。miR-1253被hsa_circ_0087862吸附。NSCLC组织中miR-1253的表达与hsa_circ_0087862呈负相关（<0.001）。miR-１253直接抑制NSCLC中RAB3D的表达。miR-1253下调或RAB3D过表达显著逆转了hsa_circ_0087862下调诱导的NSCLC细胞表型。hsa_circ_0087862下调显著抑制体内肿瘤生长（<0.01）。在异种移植瘤组织中，hsa_circ_0087862下调明显降低RAB3D、Ki67的表达并增加凋亡。
hsa_circ_0087862通过靶向miR-1253/RAB3D在NSCLC中发挥癌基因作用。
