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环状RNA hsa_circ_0087862通过靶向miR-1253/RAB3D轴在非小细胞肺癌中发挥癌基因作用。

CircRNA hsa_circ_0087862 Acts as an Oncogene in Non-Small Cell Lung Cancer by Targeting miR-1253/RAB3D Axis.

作者信息

Li Lin, Wan Ke, Xiong Linkai, Liang Shuang, Tou Fangfang, Guo Shanxian

机构信息

Department of Thoracic Oncology, Jiangxi Cancer Hospital, Nanchang 330029, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Apr 3;13:2873-2886. doi: 10.2147/OTT.S243533. eCollection 2020.

Abstract

PURPOSE

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).

METHODS

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.

RESULTS

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.

CONCLUSION

hsa_circ_0087862 acted as an oncogene in NSCLC by targeting miR-1253/RAB3D.

摘要

目的

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。

方法

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。

结果

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。

结论

hsa_circ_0087862通过靶向miR-1253/RAB3D在NSCLC中发挥癌基因作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff16/7138622/15cea16dbbad/OTT-13-2873-g0001.jpg

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