FU Yangyang,HUANG Xiaoying,WANG Liangxing. AKAP9 promotes lung adenocarcinoma cell A549 proliferation and migration via enhancing the activity of ERK pathway[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2022, 52(5): 365-370.
Abstract：Objective: To investigate the effect of A-kinase anchoring protein 9 (AKAP9) on lung cancer cell A549 and its possible molecular mechanism. Methods: Real-time quantitative polymerase chain reaction (RTqPCR) and Western blot were used to detect the knockdown effect of shRNA on lung cancer cell A549. 3-(4, 5-dimethylthiazole-2)-2, 5-diphenyltetrazolium bromide (MTT) and clone formation assay were performed to examine the role of AKAP9 in lung cancer cell proliferation. Transwell assay was performed to examine the function of AKAP9 in the migration of lung cancer cell A549. Flow cytometry was used to monitor cell cycle.Western blot was used to examine the expression of cell cycle associated proteins P27, Cyclin D1, cyclindependent kinase 4 (CDK4) and the phosphorylation level of epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase (ERK). Results: RT-qPCR and Western blot showed that shRNA could effectively silence the expression of AKAP9 in lung cancer cell A549. MTT and colony formation results showed that silencing AKAP9 significantly inhibited the proliferation and clonogenic ability of cancer cell A549 compared with the control group. Transwell assay showed that silencing AKAP9 suppressed the migration in lung cancer cell A549 compared with the control group. Flow cytometry results showed that silencing AKAP9 induced G1-S arrest in lung cancer cell A549 compared with the control group. Western blot results showed that silencing AKAP9 increased the expression of P27, decreased the expression of Cyclin D1, CDK4, and reduced the phosphorylation level of EGFR and ERK. Conclusion: Silencing AKAP9 may suppress proliferation and migration of lung cancer cell A549 by inhibiting the activity of ERK pathway.
