en زیست شناسی سلولی Cell Biology مقالات اصلی Original Article <div style="text-align: justify;"><strong><em>Background: </em></strong>Focal adhesions (FAs) are highly dynamic complex structures that assembled and&nbsp;disassembled on an ongoing basis. The balance between the two processes mediates various aspects of cell&nbsp;behavior, ranging from cell adhesion to cell migration. Assembly and disassembly processes of FAs are<br> regulated by a variety of cellular signaling proteins and adaptors. We previously demonstrated that local&nbsp;levels of Phosphatidylinositol 4,5‐bisphosphate (PtdIns(4,5)P2) in MDA-MB-231 cells increases during FA&nbsp;assembly and declines during disassembly. In this study we aimed to investigate whether PtdIns(4,5)P2&nbsp;regulates FA turnover.<br> <br> <strong><em>Methods:</em></strong> MDA-MB-231 cells were co-transfected with a labeling vinculin (or zyxin) and the PLC𝛅1-PH&nbsp;biosensor to visualize FA localization and PtdIns(4,5)P2 in the cell membrane. We also used&nbsp;pharmacological inhibitors to determine the mechanism underlying the changes of PtdIns(4,5)P2 level<br> during FA turnover and cell migration. Immunostaining, immunoprecipitation, and Western blotting were&nbsp;used to examine the localization and interaction between phospholipase C (PLC)/phosphatidylinositol 3-kinase (PI3K) FA proteins.<br> <br> <strong><em>Results: </em></strong>We showed that inhibition of PLC, PI3K significantly reduced the decline of PtdIns(4,5)P2 levels&nbsp;within FA disassembly and the slowdown rate of FA turnover and cell migration. We also showed that the&nbsp;inhibition of enzymes implicated in the downstream pathway of PtdIns(4,5)P2, such as diacylglycerol kinase&nbsp;(DAGK) and protein kinase C (PKC) significantly reduced FA turnover time and the speed of cell migration.&nbsp;Additionally, we demonstrated that PLC but not PI3K interact with FAs. In conclusion,<br> <br> <strong><em>Conclusions:</em></strong> This study suggests that dynamical changes of PtdIns(4,5)P2 might regulate FA turnover and&nbsp;facilitate cell migration.</div> Cell Migration, Focal Adhesion Turnover, MDA-MB-231 Breast Cancer Cell Line, Ptdins(4,5)P2, PLC, PI3K. 270 281 http://rbmb.net/browse.php?a_code=A-10-749-6&slc_lang=en&sid=1 Dhurgham Al-Fahad dhurgham.alfahad@sci.utq.edu.iq 100319475328460012213 100319475328460012213 Yes Department of Pharmaceutical Sciences, College of Pharmacy University of Thi-Qar, Iraq Firas Alyaseen 100319475328460012214 100319475328460012214 No Department of Pharmaceutical Sciences, College of Pharmacy University of Thi-Qar, Iraq. Ahmed Al-Amery 100319475328460012215 100319475328460012215 No Faculty of Education, Soran University, Erbil, Kurdistan Region, Iraq. Clementino Ibeas Bin 100319475328460012216 100319475328460012216 No Ilumix Biotech Ltd, London, United Kingdom.