Mechanistic study of the role of Pleckstrin-2 in glycolysis, migration and invasion of breast cancer cells

Breast cancer (BC) is the most commonly diagnosed cancer in women globally, highlighting the need for new therapeutic targets. This study aims to clarify the role of pleckstrin-2 (PLEK2) in BC progression. We particularly focus on how the transcription factor Homeobox D9 (HOXD9) regulates PLEK2 and its influence on the AKT signaling pathway. We employed quantitative PCR, Western blotting, gene knockdown and overexpression techniques, scratch assays, Transwell assays, and biochemical analyses to investigate the effects of PLEK2 on cell migration, invasion, and metabolic activity in BC cells. We found that PLEK2 was significantly overexpressed in BC tissues compared to normal tissues in the database, and this overexpression was associated with poor prognosis. Functional assays showed the modulatory effect of PLEK2 on glycolytic activity, migration, and invasion of breast cancer cells. Mechanistically, PLEK2 regulates the PI3K/AKT/mTOR pathway, HIF-1α protein levels, and the expression of its target glycolytic genes, thereby promoting glycolysis critical for cancer progression. Notably, HOXD9 was identified as a key regulator of PLEK2 expression, confirmed by luciferase assays and ChIP-qPCR. Overexpression of HOXD9 restored the migratory and invasive abilities of PLEK2-silenced cells, indicating a significant interplay between HOXD9 and PLEK2 in breast cancer. In conclusion, our study highlights the pivotal role of PLEK2 in BC cell dynamics and introduces HOXD9 as a novel regulator of PLEK2. Targeting the HOXD9/PLEK2/AKT signaling axis may offer a promising therapeutic strategy for treating breast cancer.