Background: Lung cancer is the leading cancer-related death worldwide. Patients with lung cancer mainly died of tumor metastasis and invasion. Protein kinase CK2 is an ubiquitous serine/threonine protein kinase and is frequently upregulated in various human tumors. This study aims to explore the effect and molecular mechanism of the invasion and migration of lung adenocarcinoma A549 cells after knock-down of CK2α expression.
Methods: The pSilencerTM 4.1-siCK2α-eGFP of lentiviral-mediated shRNA was constructed. The expression of CK2α was knock-downed, and a stable A549 cell line was established. The invasion and migration of A549 cell line was detected through Transwell and Boyden chamber assays. The protein expression of the PI3K/Akt signaling pathway and mesenchymal-to-epithelial transition (EMT) was evaluated using Western blot analysis.
Results: The invasion and migration of A549 cells were significantly inhibited after the knockdown of CK2α expression compared with that in the control group. p-PTEN, Akt, p-Akt473, p-Akt308, p-PDK1, p-c-Raf, and p-GSK-3β were significantly downregulated, whereas PTEN was upregulated. Moreover, vimentin, β-catenin, Snail, MMP2, and MMP9 were significantly downregulated after reducing the CK2α expression.
Conclusions: CK2α might regulate the invasion and migration of A549 cells through the PI3K/Akt/GSK-3β/Snail signaling pathway, which controls EMT in lung adenocarcinoma. .
背景与目的 肺癌已成为全球癌症死亡的首要原因，而侵袭和转移是导致肿瘤死亡的主要原因之一，蛋白激酶CK2是一种高度保守信使非依赖性丝氨酸苏氨酸蛋白激酶，其在各种肿瘤中高表达。本研究旨在探讨下调CK2α基因表达对肺腺癌A549细胞侵袭迁移的影响以及可能的机制。方法 构建pSilencerTM 4.1-shCK2α-eGFP慢病毒表达载体，建立稳定干扰CK2α表达的A549细胞株。利用Transwell和Boyden小室实验检测干扰CK2α表达前后A549细胞的侵袭及迁移的能力。Western blot检测PI3K/Akt信号通路和上皮-间充质转化（mesenchymal-to-epithelial transition, EMT）相关蛋白的表达。结果 与对照组相比，干扰CK2α表达后肺腺癌A549细胞的侵袭及迁移能力明显下降，p-PTEN、Akt、p-Akt473、p-Akt308、 p-PDK1、p-c-Raf、p-GSK-3β蛋白明显下调，PTEN蛋白表达水平显著上调。上皮-间充质转化的相关蛋白E-cadherin蛋白表达水平显著上调，而Vimentin、β-catenin、Snail蛋白表达水平显著下调，与侵袭转移相关蛋白的MMP2、MMP9表达水平显著下调。结论 CK2α可能通过PI3K/Akt/GSK-3β/Snail信号通路来调控上皮-间充质转化参与肺腺癌A549细胞的侵袭及迁移。.