Zhang Lei, Wu Yuanzhou, Hou Chunyang, Li Fangzhi
Department of Diagnostics of Traditional Chinese Medicine, Basic Medical College, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Department of Thoracic Surgery, Zhujiang Hospital of Southern Medical University, Guangzhou, Guangdong, China.
J Gene Med. 2022 May;24(5):e3414. doi: 10.1002/jgm.3414. Epub 2022 Feb 15.
Non-small cell lung cancer (NSCLC) is an aggressive tumor that accounts for a large proportion of cancer-related deaths. Cisplatin (CDDP) has been utilized to treat NSCLC. However, the efficacy of CDDP is usually restrained owing to the development of drug resistance. This study aims to reveal the molecular mechanism of the resistance of NSCLC cells to CDDP.
The expression levels of circRNA_0072088 (circ_0072088), microRNA-944 (miR-944), and LIM and SH3 protein 1 (LASP1) were measured by quantitative real-time PCR (qRT-PCR) in CDDP-resistant NSCLC tissues and cells. Protein expression was determined by Western blotting in CDDP-resistant NSCLC tissues and cells. The functional effects of circ_0072088, miR-944, and LASP1 on CDDP sensitivity and NSCLC progression were revealed by Cell Counting Kit-8 (CCK-8), flow cytometry, cell colony formation, wound healing, and transwell invasion assays. The binding relationship between miR-944 and circ_0072088 or LASP1 was identified by a dual-luciferase reporter and RNA immunoprecipitation assay. The effects of circ_0072088 knockdown on tumor growth in vivo were analyzed by an in vivo tumor formation assay.
Circ_0072088 and LASP1 expression were significantly upregulated, while miR-944 expression was downregulated in CDDP-resistant NSCLC tissues and cells as compared with control groups. Circ_0072088 expression was significantly associated with tumor-node-metastasis stage and tumor size. Functionally, circ_0072088 knockdown improved CDDP sensitivity and repressed NSCLC cell malignancy, whereas miR-944 inhibitor hindered these effects. Mechanistically, circ_0072088 functioned as a sponge for miR-944 and miR-944 targeted LASP1. Circ_0072088 knockdown improved the sensitivity of tumor to CDDP in vivo.
Circ_0072088 silencing improved CDDP sensitivity and inhibited NSCLC progression by downregulating LASP1 expression through sponging miR-944. These data provide novel insight into the resistance of NSCLC to CDDP.
非小细胞肺癌（NSCLC）是一种侵袭性肿瘤，在癌症相关死亡中占很大比例。顺铂（CDDP）已被用于治疗NSCLC。然而，由于耐药性的产生，CDDP的疗效通常受到限制。本研究旨在揭示NSCLC细胞对CDDP耐药的分子机制。
通过定量实时PCR（qRT-PCR）检测顺铂耐药的NSCLC组织和细胞中环状RNA_0072088（circ_0072088）、微小RNA-944（miR-944）和LIM和SH3结构域蛋白1（LASP1）的表达水平。通过蛋白质印迹法测定顺铂耐药的NSCLC组织和细胞中的蛋白质表达。通过细胞计数试剂盒-8（CCK-8）、流式细胞术、细胞集落形成、伤口愈合和Transwell侵袭实验揭示circ_0072088、miR-944和LASP1对CDDP敏感性和NSCLC进展的功能影响。通过双荧光素酶报告基因和RNA免疫沉淀实验确定miR-944与circ_0072088或LASP1之间的结合关系。通过体内肿瘤形成实验分析circ_0072088敲低对体内肿瘤生长的影响。
与对照组相比，顺铂耐药的NSCLC组织和细胞中circ_0072088和LASP1表达显著上调，而miR-944表达下调。Circ_0072088表达与肿瘤-淋巴结-转移分期和肿瘤大小显著相关。在功能上，circ_0072088敲低提高了CDDP敏感性并抑制了NSCLC细胞的恶性程度，而miR-944抑制剂则阻碍了这些作用。机制上，circ_0072088作为miR-944的海绵，而miR-944靶向LASP1。Circ_0072088敲低提高了肿瘤在体内对CDDP的敏感性。
Circ_0072088沉默通过海绵化miR-944下调LASP1表达，从而提高CDDP敏感性并抑制NSCLC进展。这些数据为NSCLC对CDDP的耐药性提供了新的见解。
