Dhurgham Al-Fahad - Department of Pharmaceutical Sciences, College of Pharmacy University of Thi-Qar, Thi-Qar ۶۴۰۰۱, Iraq.
Firas Alyaseen - Department of Pharmaceutical Sciences, College of Pharmacy University of Thi-Qar, Thi-Qar ۶۴۰۰۱, Iraq.
Ahmed Al-Amery - epartment of Physiology, College of Medicine, University of Thi-Qar, Iraq.
Gagandeep Singh - Viral Research and Diagnostic Laboratory, Department of Microbiology, Osmanian Medical College, Hyderabad, Telangana, India & Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, New Delhi, India.
Mote Srinath - Viral Research and Diagnostic Laboratory, Department of Microbiology, Osmanian Medical College, Hyderabad, Telangana, India.
Yahya Abbas - Department of Biology, College of Science, University of Thi-Qar, Thi-Qar, ۶۴۰۰۱ Iraq.
Hafiz Muzzammel Rehman - School of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan.
Yahya Abbas - Department of Biology, College of Science, University of Thi-Qar, Thi-Qar, ۶۴۰۰۱ Iraq.

Background: The assembly and disassembly of the focal adhesions (FA) components occurs throughout life cycle of adhesion, with conservation of balance between removal and recruitment rate during temporal stages. Previous studies have demonstrated that phosphotidyilinositols play a role in regulating FA turnover. However, a little attention has been given to quantify the dynamics changes of Phosphatidylinositol ۳,۴,۵-trisphosphate (PtdIns (۳,۴,۵) P۳) within and during fast and slow turnover rates of FA. Methods: MDA-MB-۲۳۱ breast cancer cell line was used as a model in this study due to high metastatic and motile. These cells were co-transfected with GFP- paxillin/vinculin, as FA marker, and the GFP/mCherry-Btk-PH, as a biosensor to visualize PtdIns (۳,۴,۵) P۳. Confocal time-lapse images were used to monitor changes or differences in the local generation of PtdIns (۳,۴,۵) P۳ within and during assembly and disassembly of FA. Following transfection, immunostaining was used to examine the spatial colocalization between FA and PtdIns (۳,۴,۵) P۳. Results: Our data demonstrated that PtdIns (۳,۴,۵) P۳ co-localized with FAs and increase during assembly and decline during disassembly of FA which exhibits slow turnover rates and was in a constant level during assembly and disassembly of FA that displays fast turnover rates.  Conclusions: Our result suggested that the dynamic changes of PtdIns (۳,۴,۵) P۳, it may depend on components undergo turnover, such that early, nascent FA displays fast turnover rates and mature FA exhibits slow turnover rates. Thus, the local enrichment of PtdIns (۳,۴,۵) P۳ enhances FA assembly and disassembly activation.

Cancer cell migration, Focal adhesion turnover, MDA-MB-۲۳۱ cell line, PtdIns (۳, ۴, ۵) P۳.