Effects of phosphatidylinositol 4-phosphate on invasion and metastasis of Human glioma cell U87

Abstract

Objective We detected the expression of phosphatidylinositol 4-phosphate (PI4P) after overexpression or silencing PI4Pby short hairpin RNA (shRNA) in human glioma cells U87. We discussed the role ofPI4P on human glioma migration through acting U87 by overexpression or silencing PI4P. To explore PI4P mechanism during the process of human glioma. Methods LV-Helper1, LV-Helper2, pWPXLd-PI4P or pLL3.7-shPI4P construct and product Lentivirus. The lentivirus of pWPXLd-puro, pWPXLd-PI4P, pLL3.7-Scramble and pLL3.7-shPI4P transfected U87and established the stable cell lines.The migratory and invasive ability were tested by wound, healing assay and transwell invasion assay respectively. Expressions of PI4Pwere observed by Western blotting(WB). Results We have successfully packaged lentivirus of pWPXLd-puro, pWPXLd-PI4P, pLL3.7-Scramble and pLL3.7-shPI4P. Establishment of stable U87-GFP, U87-GFP-PI4P, U87- Scramble and U87-shPI4P cell lines. Overexpression of PI4P , the invasion number of U87-GFP-PI4P cell line is (197.69±2.08), while the invasion number of U87-GFP cell line is (120.00±2.64); the migratory ability numbers of U87-GFP-PI4P and U87-GFPcell lines are (70.00±0.58), (50.00±1.02) respectively (t=39.960, P=0.000 and t=33.150, P=0.000). Silencing of PI4P , the invasive ability of U87-GFP-PI4P and U87-GFP cell lines are (79.67±1.53), (117.00±2.00), while the migratory ability numbers are (19.33±2.08), (46.67±1.52), respectively (t=25.700, P=0.000 and t=18.340, P=0.000). Overexpression of PI4P promoted the migratory invasive ability of human glioma cells and the expressions of RhoA (t=17.270, P=0.000). Silencing of PI4P inhibited themigratory invasive ability of human glioma cells and the expressions of RhoA (t=35.120, P=0.000). Conclusion PI4P promotesmigratory invasive ability of human golima U87 cells, its downstream protein RhoA were also involved in the regulation of this process. Key words: Glioma; Migration; U87; Phosphatidylinositol 4-phosphate; RhoA