Circ_0074027 contributes to non-small cell lung cancer progression through positively modulating RHOA via sequestering miR-2467-3p
Circ_0074027 contributes to non-small cell lung cancer progression through positively modulating RHOA via sequestering miR-2467-3p
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DOI：
10.1007/s10863-021-09876-6
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发表时间：
2021-03-09
影响因子：
3
通讯作者：
Liu, Ying
中科院分区：
文献类型：
--
作者：
Duan, Zhihui;Wei, Shuqing;Liu, Ying
非小细胞肺癌（NSCLC）是一种常见癌症，5年生存率不佳。我们打算探讨环状 RNA_0074027 (circ_0074027) 在 NSCLC 进展中的作用。通过定量实时聚合酶链式反应 (qRT-PCR) 测定 circ_0074027、其线性形式的信使 RNA (mRNA) 的水平，如同源结构域 1 (PITX1)、microRNA-2467-3p (miR-2467-3p) 和 ras 同源家族成员 A (RHOA) mRNA。进行细胞计数试剂盒8（CCK8）测定和平板集落测定来测量NSCLC细胞的增殖能力。 Transwell实验用于评估细胞迁移和侵袭能力。采用流式细胞术分析细胞凋亡率。使用双荧光素酶报告基因测定和RNA免疫沉淀(RIP)测定来测试miR-2467-3p与circ_0074027或RHOA之间的相互作用。采用蛋白质印迹法评估 NSCLC 细胞中 RHOA 的蛋白水平。建立小鼠异种移植模型来评估circ_0074027在体内肿瘤生长中的作用。 Circ_0074027 表达在 NSCLC 组织和细胞系中显着升高。 Circ_0074027 敲低或 miR-2467-3p 过表达抑制细胞增殖、迁移和侵袭，并促进 NSCLC 细胞的细胞凋亡。 Circ_0074027 与 miR-2467-3p 相互作用，RHOA 是 NSCLC 细胞中 miR-2467-3p 的靶标。 RHOA 沉默阻断了 NSCLC 细胞的恶性潜能。 Circ_0074027沉默主要通过上调miR-2467-3p抑制NSCLC细胞的恶性表型。 Circ_0074027 敲低显着阻止了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 表达来加速 NSCLC 进展。
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.