Department of Cancer Radiotherapy, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China (Anhui Provincial Cancer Hospital), Hefei, Anhui, China.
Department of Science and Education, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China (Anhui provincial Cancer Hospital), Hefei, Anhui, China.
J Clin Lab Anal. 2022 May;36(5):e24304. doi: 10.1002/jcla.24304. Epub 2022 Mar 21.
Chemoresistance is one of the major obstacles for tumor treatment. Circular RNAs (circRNAs) have been confirmed to play vital roles in chemoresistance of cancer, including esophageal squamous cell carcinoma (ESCC). We investigated the roles and mechanisms of circ_0007142 in cisplatin (DDP) resistance of ESCC.
Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to determine the levels of circ_0007142, DOCK1 mRNA, microRNA-494-3p (miR-494-3p) and LIM And SH3 Protein 1 (LASP1) mRNA. RNase R assay was conducted to analyze the characteristic of circ_0007142. Cell Counting Kit-8 (CCK-8) assay was performed to evaluate IC50 of DDP. Flow cytometry analysis, 5-ethynyl-2'-deoxyuridine (EdU) assay and transwell assay were carried out to examine cell apoptosis, proliferation and invasion, respectively. Dual-luciferase reporter assay was employed to verify the association between miR-494-3p and circ_0007142 or LASP1. Murine xenograft assay was conducted to investigate the role of circ_0007142 in DDP resistant in vivo. The protein level of LASP1 in tumors was measured by Immunohistochemistry (IHC) analysis.
Circ_0007142 was upregulated in DDP-resistant ESCC tissues and cells. Circ_0007142 knockdown improved DDP sensitivity, induced cell apoptosis and hampered cell proliferation and invasion in DDP-resistant ESCC cells. Circ_0007142 functioned as the sponge for miR-494-3p and miR-494-3p inhibition reversed the impacts of circ_0007142 knockdown on DDP resistance, cell apoptosis, proliferation, and invasion. LASP1 was a target of miR-494-3p, and the effects on DDP resistance, cell apoptosis, growth, and invasion mediated by LASP1 downregulation were rescued by miR-494-3p inhibition. Moreover, circ_0007142 knockdown enhanced DDP sensitivity in vivo.
Circ_0007142 improved DDP resistance of ESCC by upregulating LASP1 via sponging miR-494-3p.
化疗耐药是肿瘤治疗的主要障碍之一。环状 RNA（circRNA）已被证实在癌症的化疗耐药中发挥重要作用，包括食管鳞状细胞癌（ESCC）。我们研究了 circ_0007142 在 ESCC 顺铂（DDP）耐药中的作用和机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 circ_0007142、DOCK1mRNA、微小 RNA-494-3p（miR-494-3p）和 LIM 和 SH3 蛋白 1（LASP1）mRNA 的水平。采用 RNase R 分析检测 circ_0007142 的特征。采用细胞计数试剂盒-8（CCK-8）检测 DDP 的 IC50。采用流式细胞术分析、5-乙炔基-2'-脱氧尿苷（EdU）检测和 Transwell 检测分别评估细胞凋亡、增殖和侵袭。采用双荧光素酶报告基因检测验证 miR-494-3p 与 circ_0007142 或 LASP1 的关联。采用小鼠异种移植实验研究 circ_0007142 在体内对 DDP 耐药的作用。采用免疫组织化学（IHC）分析检测肿瘤中 LASP1 的蛋白水平。
circ_0007142 在 DDP 耐药的 ESCC 组织和细胞中上调。circ_0007142 敲低可提高 DDP 敏感性，诱导 DDP 耐药的 ESCC 细胞凋亡，抑制细胞增殖和侵袭。circ_0007142 作为 miR-494-3p 的海绵，抑制 miR-494-3p 可逆转 circ_0007142 敲低对 DDP 耐药、细胞凋亡、增殖和侵袭的影响。LASP1 是 miR-494-3p 的靶基因，下调 LASP1 介导的 DDP 耐药、细胞凋亡、生长和侵袭的作用可被 miR-494-3p 抑制所挽救。此外，circ_0007142 敲低可增强体内 DDP 的敏感性。
circ_0007142 通过海绵吸附 miR-494-3p 上调 LASP1 可提高 ESCC 的 DDP 耐药性。
