Department of Thoracic Surgery, Cixi People's Hospital, Cixi City, China.
Department of Respiratory Medicine, Cixi People's Hospital, Cixi City, China.
J Clin Lab Anal. 2021 Aug;35(8):e23865. doi: 10.1002/jcla.23865. Epub 2021 Jul 27.
Lung cancer is the most commonly diagnosed cancer and leading cause of cancer death, with 80%-85% of non-small cell lung cancer (NSCLC). Circular RNAs (circRNAs) have been shown to be promising early diagnostic and therapeutic molecular biomarkers for NSCLC. However, biological role and regulatory mechanism of circRNA WHSC1 (circWHSC1) in NSCLC are unknown. Therefore, we aim to explore the function and mechanism of circWHSC1 in NSCLC oncogenesis and progression.
qRT-PCR was used for circWHSC1 level evaluation; Kaplan-Meier was used for survival analysis; bioinformatics, dual-luciferase activity, and RNA pull-down were used for evaluating competing endogenous RNA (ceRNA) network; cell viability, colony formation, apoptosis, migration, and invasion were used for cell function analysis; function gain and loss with rescue experiments were used for exploring mechanism of circWHSC1 in NSCLC development.
Significantly up-regulated circWHSC1 and down-regulated microRNA-296-3p (miR-296-3p) were identified in NSCLC tissues and cells. Up-regulated circWHSC1 was associated with poor prognosis in NSCLC patients. MiR-296-3p was sponged by circWHSC1, and AKT serine/threonine kinase 3 (AKT3) was target of miR-296-3p; meanwhile, miR-296-3p over-expression significantly down-regulated AKT3 expression, and co-transfecting anti-miR-296-3p rescued circWHSC1 silence caused AKT3 down-regulation. CircWHSC1 silence significantly inhibited colony formation, viability, invasion, and migration, while increased NSCLC cell apoptosis, which were partially rescued by anti-miR-296-3p.
CircWHSC1 is an independent indicator of poor prognosis in NSCLC patients, and functions as a ceRNA of miR-296-3p to up-regulate AKT3, consequently promotes NSCLC cell growth and metastasis. Targeting circWHSC1 might be a prospective strategy for diagnosis, therapeutics, and prognosis of NSCLC.
肺癌是最常见的癌症诊断和癌症死亡的主要原因，其中 80%-85%是非小细胞肺癌（NSCLC）。环状 RNA（circRNA）已被证明是 NSCLC 的有前途的早期诊断和治疗分子生物标志物。然而，环状 RNA WHSC1（circWHSC1）在 NSCLC 中的生物学作用和调节机制尚不清楚。因此，我们旨在探讨 circWHSC1 在 NSCLC 发生和发展中的功能和机制。
qRT-PCR 用于评估 circWHSC1 水平；Kaplan-Meier 用于生存分析；生物信息学、双荧光素酶活性和 RNA 下拉用于评估竞争性内源性 RNA（ceRNA）网络；细胞活力、集落形成、凋亡、迁移和侵袭用于细胞功能分析；通过功能增益和缺失的挽救实验来探讨 circWHSC1 在 NSCLC 发展中的机制。
在 NSCLC 组织和细胞中发现显著上调的 circWHSC1 和下调的 microRNA-296-3p（miR-296-3p）。上调的 circWHSC1 与 NSCLC 患者的不良预后相关。circWHSC1 可作为 miR-296-3p 的海绵，AKT 丝氨酸/苏氨酸激酶 3（AKT3）是 miR-296-3p 的靶标；同时，过表达 miR-296-3p 显著下调 AKT3 表达，共转染抗 miR-296-3p 可挽救 circWHSC1 沉默引起的 AKT3 下调。circWHSC1 沉默显著抑制 NSCLC 细胞集落形成、活力、侵袭和迁移，同时增加细胞凋亡，而过表达 miR-296-3p 可部分挽救。
circWHSC1 是 NSCLC 患者不良预后的独立指标，作为 miR-296-3p 的 ceRNA 上调 AKT3，进而促进 NSCLC 细胞生长和转移。靶向 circWHSC1 可能是 NSCLC 诊断、治疗和预后的有前景的策略。
