Phosphorylase kinase regulatory subunit alpha 1 as a novel biomarker involved in development and progression of lung cancer: comprehensive bioinformatic analysis and experiment validation.

Abstract

Lung cancer remains a critical global health issue, with the molecular intricacies and specific role of phosphorylase kinase regulatory subunit alpha 1 (PHKA1) not well understood. This study aims to utilize an integrated bioinformatic analyses and experimental validations to investigate PHKA1 in lung cancer. Bioinformatic tools provided a pan-cancer perspective, emphasizing PHKA1's oncogenic potential such as GEPIA and UALCAN revealed significantly elevated mRNA expression of PHKA1 in lung adenocarcinoma (LUAD) across various stages, highlighting its diagnostic significance. The study also explored the impact of PHKA1 on immune cells in LUAD, finding strong correlations between high PHKA1 expression and increased presence of CD4+, CD8+ T cells, and macrophages using TIMER 2.0 database. Survival analysis using Kaplan-Meier (KM) plots showed that patients with elevated PHKA1 levels had poorer prognoses, reinforcing its potential as an oncogenic gene in non-small cell lung cancer (NSCLC) by KM plotter. Immunohistochemistry by HPA database supported these findings, demonstrating increased PHKA1 protein levels in lung cancer tissues. Protein-protein interaction analysis using STRING and cytoscape identified a network of genes linked to PHKA1 in NSCLC, offering insights into its functional interactions. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis suggested that PHKA1 might be involved in the PI3K-AKT signaling pathway, known for its role in cell growth and survival, further supporting its oncogenic role. Experimental validation through RT-PCR and Western blot confirmed elevated PHKA1 expression in A549 and NCI-H460 lung cancer cells. Moreover, deletion of PHKA1 reduces cell growth and promotes apoptosis in A549 cells and confirmed by Western blot. In conclusion, this comprehensive study suggests that targeting PHKA1 in NSCLC could be a promising therapeutic strategy, highlighting its crucial role in the molecular landscape of lung cancer progression.