ABSTRACT
Introduction: Phosphoinositides play a key role in the regulation of focal adhesions (FAs) turnover during cell adhesion and migration. However, their potential role in FA turnover at leading and trailing edge of cell are not yet fully understood. In this study, we investigate their spatial co-localisation with paxillin directly at leading and trailing edge of MDA-MB-231 breast cancer cell line. Materials and methods: Cell lines and cell culture experiments were done using MDA-MB-231human adenocarcinoma cells. Co-trnasfection and confocal microscopy were performed to visualise phosphoinositides and FAs by using GFP-C1-PLCdelta-PH/Btk-PH-GFP and 3 µɡ paxillin-RFP as biosensors. Then, ImageJ was used to measure co-localisation point between Phosphatidylinositol 4,5-trisphosphate (PtdIns(4,5)P2) or Phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) and paxillin Spearman's rank correlation value was taken between PtdIns(4,5)P2/PtdIns(3,4,5)P3. Results: Our results demonstrate that the spatial co-localistion of PtdIns(4,5)P2 and PtdIns(3,4,5)P3 with FA at leading and trailing age of cell were slightly changed. Conclusions: This suggests that PtdIns(4,5)P2 and PtdIns(3,4,5)P3 play an equal role at the leading and trailing edges of the cancer metastasis through interaction with FA proteins (AU)
