环状 RNA HIPK2 通过促进血管生成和化疗耐药性促进非小细胞肺癌的恶性进展。

The malignant property of circHIPK2 for angiogenesis and chemoresistance in non-small cell lung cancer.

机构信息

Department of Thoracic Surgery, Cangzhou Central Hospital, Cangzhou City, Hebei Province, 061000, China.

出版信息

Exp Cell Res. 2022 Oct 15;419(2):113276. doi: 10.1016/j.yexcr.2022.113276. Epub 2022 Jul 19.

DOI:10.1016/j.yexcr.2022.113276

Abstract

Chemotherapy resistance limits the efficacy of cisplatin (DDP) when treating non-small cell lung cancer (NSCLC). Circular RNAs (circRNAs) confers a regulatory role in drug resistance. Innovatively, the regulatory role of circular RNA HIPK2 (circHIPK2) in DDP resistance was probed in the work. In this research, tumor tissues and matched normal tissues were obtained from 52 NSCLC patients, and the expressions of circHIPK2, miR-1249-3p and VEGFA in the tissues were detected by qPCR or Western Blot. Correlation analysis of circHIPK2 expression with survival prognosis and clinicopathological features was conducted. Parental NSCLC cell lines (A549, H460) and DDP-resistant cell lines (A549/DDP, H460/DDP) were selected, and the expression of circHIPK2, miR-1249-3p and VEGFA in the cells were detected. Cell IC50 value, proliferation, migration, invasion, apoptosis and angiogenesis were detected. Tumor xenografts were established to detect the role of circHIPK2 in vivo. The binding relationship between circHIPK2, miR-1249-3p and VEGFA was verified by dual luciferase reporter experiment, RNA pull down and RIP experiment. Our data showed that circHIPK2 and VEGFA were abnormally overexpressed and miR-1249-3p was underexpressed in DDP-resistant NSCLC tissues and cell lines. CircHIPK2 knockdown or miR-1249-3p upregulation inhibited DDP resistance, malignant behavior, and angiogenesis in NSCLC. CircHIPK2 by competitive absorption of miR-1249-3p mediated VEGFA. CircHIPK2 promoted the sensitivity of drug-resistant cells to DDP in NSCLC by regulating VEGFA. CircHIPK2 enhanced the growth of DDP-resistant NSCLC cells in vivo. In conclusion, circHIPK2 has the malignant property for angiogenesis and chemoresistance in NSCLC via the network of miR-1249-3p/VEGFA.

摘要

化疗耐药限制了顺铂（DDP）治疗非小细胞肺癌（NSCLC）的疗效。环状 RNA（circRNA）在耐药中发挥调节作用。在这项工作中，创新性地研究了环状 RNA HIPK2（circHIPK2）在 DDP 耐药中的调节作用。在这项研究中，从 52 名 NSCLC 患者中获得肿瘤组织和匹配的正常组织，并通过 qPCR 或 Western Blot 检测组织中 circHIPK2、miR-1249-3p 和 VEGFA 的表达。进行 circHIPK2 表达与生存预后和临床病理特征的相关性分析。选择亲本 NSCLC 细胞系（A549、H460）和 DDP 耐药细胞系（A549/DDP、H460/DDP），并检测细胞中 circHIPK2、miR-1249-3p 和 VEGFA 的表达。检测细胞 IC50 值、增殖、迁移、侵袭、凋亡和血管生成。建立肿瘤异种移植模型以检测 circHIPK2 在体内的作用。通过双荧光素酶报告实验、RNA 下拉和 RIP 实验验证 circHIPK2、miR-1249-3p 和 VEGFA 之间的结合关系。我们的数据表明，circHIPK2 和 VEGFA 在 DDP 耐药 NSCLC 组织和细胞系中异常过表达，而 miR-1249-3p 表达下调。circHIPK2 敲低或 miR-1249-3p 上调抑制 NSCLC 中的 DDP 耐药、恶性行为和血管生成。circHIPK2 通过竞争性吸收 miR-1249-3p 介导 VEGFA。circHIPK2 通过调节 VEGFA 增强 NSCLC 耐药细胞对 DDP 的敏感性。circHIPK2 增强了体内 DDP 耐药 NSCLC 细胞的生长。总之，circHIPK2 通过 miR-1249-3p/VEGFA 网络在 NSCLC 中具有血管生成和化疗耐药的恶性特性。