Department of Radiotherapy, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, Shanxi, China.
Department of Cardiology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, Shanxi, China.
Mol Biotechnol. 2023 Sep;65(9):1421-1431. doi: 10.1007/s12033-023-00656-8. Epub 2023 Jan 11.
This study aimed to assess the role of cyclin-dependent kinase-like 3 (CDKL3) in the progression of non-small cell lung cancer (NSCLC) as well as the underlying mechanisms. Western blot and qRT-PCR were utilized to analyze CDKL3 expression in 30 pairs of NSCLC and paraneoplastic tissues. A549 cells with CDKL3 knockdown and PC9 cells with CDKL3 overexpression were constructed by infecting cells with lentiviruses expressing shRNA of CDKL3 and expressing a full-length CDKL3 mRNA, respectively. The CCK-8 assay, flow cytometry, wound healing assay, and Transwell assay were carried out to detect cell viability, apoptosis, migration, and invasion, respectively. Autophagosome morphology was observed by electron microscopy experiments, the expression of key components of the PI3K/Akt/mTOR pathway was examined via Western blot and their mRNA expression levels were determined. Besides, the stably infected NSCLC cells with reduced expression or overexpression of CDKL3 were inoculated into the right-back flank of mice to generate tumors. The results showed that CDKL3 expression was dramatically increased in NSCLC tissues. Moreover, CDKL3 promoted the viability and migration of NSCLC cells by suppressing autophagy in vitro and in vivo. In addition, CDKL3 might modulate PI3K/Akt/mTOR signaling in NSCLC. Overall, CDKL3 might promote NSCLC cell viability and metastasis by inhibiting autophagy and activating the PI3K/Akt/mTOR signaling pathway.
本研究旨在评估周期素依赖性激酶样 3（CDKL3）在非小细胞肺癌（NSCLC）进展中的作用及其潜在机制。通过Western blot 和 qRT-PCR 分析了 30 对 NSCLC 和癌旁组织中 CDKL3 的表达。通过感染表达 shRNA 的慢病毒和表达全长 CDKL3 mRNA 的慢病毒，构建了 CDKL3 敲低的 A549 细胞和 CDKL3 过表达的 PC9 细胞。通过 CCK-8 测定、流式细胞术、划痕愈合实验和 Transwell 实验分别检测细胞活力、细胞凋亡、迁移和侵袭。通过电子显微镜实验观察自噬体形态，通过 Western blot 检测 PI3K/Akt/mTOR 通路关键组成部分的表达，并测定其 mRNA 表达水平。此外，将 CDKL3 表达降低或过表达的稳定感染 NSCLC 细胞接种到小鼠右后背部以生成肿瘤。结果表明，CDKL3 在 NSCLC 组织中表达显著增加。此外，CDKL3 通过抑制自噬在体外和体内促进 NSCLC 细胞的活力和迁移。此外，CDKL3 可能在 NSCLC 中调节 PI3K/Akt/mTOR 信号通路。总的来说，CDKL3 可能通过抑制自噬和激活 PI3K/Akt/mTOR 信号通路来促进 NSCLC 细胞的活力和转移。
