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环状 RNA 0074027 通过海绵吸附 miR-2467-3p 正向调控 RHOA 促进非小细胞肺癌进展。

Circ_0074027 contributes to non-small cell lung cancer progression through positively modulating RHOA via sequestering miR-2467-3p.

机构信息

Department of Thoracic Surgery, Shanxi Provincial Cancer Hospital, Taiyuan, 030013, Shanxi, China.

Department of Head and Neck Radiotherapy Combined, Shanxi Provincial Cancer Hospital, No. 3 Worker's New Street, Xinhualing District, Taiyuan, 030013, Shanxi, China.

出版信息

J Bioenerg Biomembr. 2021 Apr;53(2):223-233. doi: 10.1007/s10863-021-09876-6. Epub 2021 Mar 9.

Abstract

Non-small cell lung cancer (NSCLC) is a common cancer with an unfavorable 5-year survival rate. We intended to explore the role of circular RNA_0074027 (circ_0074027) in NSCLC progression. The levels of circ_0074027, messenger RNA (mRNA) of its linear form paired like homeodomain 1 (PITX1), microRNA-2467-3p (miR-2467-3p) and ras homolog family member A (RHOA) mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK8) assay and plate colony assay were conducted to measure the proliferation ability of NSCLC cells. Transwell assays were used to assess cell migration and invasion abilities. Flow cytometry was utilized to analyze cell apoptosis rate. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were used to test the interaction between miR-2467-3p and circ_0074027 or RHOA. Western blot assay was performed to evaluate the protein level of RHOA in NSCLC cells. Murine xenograft model was built to evaluate the role of circ_0074027 in tumor growth in vivo. Circ_0074027 expression was prominently elevated in NSCLC tissues and cell lines. Circ_0074027 knockdown or miR-2467-3p overexpression suppressed cell proliferation, migration and invasion and facilitated cell apoptosis of NSCLC cells. Circ_0074027 interacted with miR-2467-3p, and RHOA was a target of miR-2467-3p in NSCLC cells. RHOA silencing blocked the malignant potential of NSCLC cells. Circ_0074027 silencing restrained the malignant phenotypes of NSCLC cells largely through up-regulating miR-2467-3p. Circ_0074027 knockdown notably blocked xenograft tumor growth in vivo. In conclusion, circ_0074027 accelerated NSCLC progression by binding to miR-2467-3p to induce RHOA expression.

摘要

非小细胞肺癌(NSCLC)是一种常见的癌症,其 5 年生存率较差。我们旨在探讨环状 RNA_0074027(circ_0074027)在 NSCLC 进展中的作用。通过实时定量聚合酶链反应(qRT-PCR)测定 circ_0074027、其线性形式配对同源域 1(PITX1)的信使 RNA(mRNA)、微小 RNA-2467-3p(miR-2467-3p)和 ras 同源家族成员 A(RHOA)mRNA 的水平。通过细胞计数试剂盒-8(CCK8)检测和平板集落检测测定 NSCLC 细胞的增殖能力。通过 Transwell 检测评估细胞迁移和侵袭能力。通过流式细胞术分析细胞凋亡率。双荧光素酶报告基因检测和 RNA 免疫沉淀(RIP)检测用于测试 miR-2467-3p 与 circ_0074027 或 RHOA 之间的相互作用。Western blot 检测用于评估 NSCLC 细胞中 RHOA 的蛋白水平。建立小鼠异种移植模型以评估 circ_0074027 在体内肿瘤生长中的作用。Circ_0074027 在 NSCLC 组织和细胞系中表达明显上调。Circ_0074027 敲低或 miR-2467-3p 过表达抑制 NSCLC 细胞的增殖、迁移和侵袭,促进细胞凋亡。Circ_0074027 与 miR-2467-3p 相互作用,RHOA 是 NSCLC 细胞中 miR-2467-3p 的靶标。RHOA 沉默阻断 NSCLC 细胞的恶性潜能。Circ_0074027 沉默通过上调 miR-2467-3p 极大地抑制 NSCLC 细胞的恶性表型。Circ_0074027 敲低显著抑制体内异种移植肿瘤的生长。总之,circ_0074027 通过与 miR-2467-3p 结合诱导 RHOA 表达加速 NSCLC 的进展。

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