环状 RNA 0011292 通过调控 miR-433-3p/CHEK1 轴抑制紫杉醇耐药非小细胞肺癌的进展和耐药性。
Circ_0011292 knockdown mitigates progression and drug resistance in PTX-resistant non-small-cell lung cancer cells by regulating miR-433-3p/CHEK1 axis.
机构信息
Department of Respiratory and Critical Care Medicine, Jingmen No.1 People's Hospital, Jingmen City, China.
Department of Reprodutive Medicine Center, Jingmen No.1 People's Hospital, Jingmen City, China.
出版信息
Thorac Cancer. 2022 May;13(9):1276-1288. doi: 10.1111/1759-7714.14378. Epub 2022 Mar 29.
BACKGROUND
Non-small-cell lung cancer (NSCLC) is one of the most common malignant tumors on earth. Circular RNAs have been disclosed to be vital regulators in the chemoresistance and development of diverse cancers, including NSCLC. Here, we attempted to explore the function of circ_0011292 in paclitaxel (PTX)-resistant NSCLC cells.
METHODS
Quantitative real-time polymerase chain reaction or western blot was performed to detect the expression of circ_0011292, microRNA-433-3p (miR-433-3p), and checkpoint kinase 1 (CHEK1). Ribonuclease R (RNase R) assay was performed to assess the stability of circ_0011292. Cell Counting Kit-8 assay was conducted to evaluate the half maximal inhibitory concentration of PTX and cell viability. Cell proliferation was monitored by Edu incorporation and colony formation assays. Cell cycle and apoptosis were detected by flow cytometry. Transwell assay was implemented to assess cell migration and invasion. Western blot assay was utilized to determine the protein levels. Dual-luciferase reporter assay was carried out to verify the targeted interaction between miR-433-3p and circ_0011292 or CHEK1. Xenograft tumor model was constructed for determining the effect of circ_0011292 in NSCLC growth in vivo.
RESULTS
Circ_0011292 was upregulated in PTX-resistant NSCLC tissues and cells. Circ_0011292 or CHEK1 knockdown enhanced PTX sensitivity and cell apoptosis, and repressed cell proliferation, migration, and invasion in PTX-resistant NSCLC cells. Mechanistically, circ_0011292 was a sponge of miR-433-3p and miR-433-3p directly targeted CHEK1. Meanwhile, silencing miR-433-3p or overexpressing CHEK1 respectively abrogated the impacts of circ_0011292 deletion or miR-433-3p introduction on PTX resistance and cell progression in PTX-resistant NSCLC cells in vitro. Moreover, circ_0011292 could positively modulate CHEK1 expression through sponging miR-433-3p. In addition, circ_0011292 knockdown retarded tumor growth of NSCLC in vivo.
CONCLUSION
Circ_0011292 could accelerate PTX resistance and cell malignant progression of NSCLC cells partially through the regulation of circ_0011292/miR-433-3p/CHEK1 axis.
背景
非小细胞肺癌（NSCLC）是地球上最常见的恶性肿瘤之一。环状 RNA 已被揭示为包括 NSCLC 在内的多种癌症的化疗耐药性和发展的重要调节剂。在这里，我们试图探讨 circ_0011292 在紫杉醇（PTX）耐药性 NSCLC 细胞中的作用。
方法
通过实时定量聚合酶链反应或 Western blot 检测 circ_0011292、微小 RNA-433-3p（miR-433-3p）和检查点激酶 1（CHEK1）的表达。核糖核酸酶 R（RNase R）试验用于评估 circ_0011292 的稳定性。细胞计数试剂盒-8 测定法用于评估 PTX 的半最大抑制浓度和细胞活力。通过 EdU 掺入和集落形成测定法监测细胞增殖。通过流式细胞术检测细胞周期和细胞凋亡。Transwell 测定法用于评估细胞迁移和侵袭。Western blot 测定法用于测定蛋白质水平。双荧光素酶报告基因测定法用于验证 miR-433-3p 与 circ_0011292 或 CHEK1 之间的靶向相互作用。构建异种移植肿瘤模型以确定 circ_0011292 在体内 NSCLC 生长中的作用。
结果
circ_0011292 在 PTX 耐药性 NSCLC 组织和细胞中上调。circ_0011292 或 CHEK1 敲低增强了 PTX 敏感性和细胞凋亡，并抑制了 PTX 耐药性 NSCLC 细胞的增殖、迁移和侵袭。机制上，circ_0011292 是 miR-433-3p 的海绵，miR-433-3p 直接靶向 CHEK1。同时，沉默 miR-433-3p 或过表达 CHEK1 分别消除了 circ_0011292 缺失或 miR-433-3p 导入对体外 PTX 耐药性和细胞进展的影响。此外，circ_0011292 通过海绵 miR-433-3p 可正向调节 CHEK1 表达。此外，circ_0011292 敲低可减缓 NSCLC 体内肿瘤的生长。
结论
circ_0011292 可通过调节 circ_0011292/miR-433-3p/CHEK1 轴，部分促进 NSCLC 细胞的 PTX 耐药性和细胞恶性进展。
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