Fig. 6: Lyso-PS activates RhoA, cooperates with HIF1A to regulate RhoA expression, promotes invasion of gastric cancer cell.
Pull-down assays and western blotting to examine the activated RhoA (GTP-RhoA). A Silencing miR-4646-5p or Abhd16a in Drosha decreased GC cells reduced the activated RhoA, then be restored by lyso-PS (10 μM) administration. B miR-4646-5p and ABHD16A overexpression in gastric cancer cells or exogenous lyso-PS (10 μM) were shown to activate RhoA activity in GC cells with endogenous GPR34, but not in GPR34-silenced or PTX (inhibitor of Gαi, 100 ng/ml, 4 h) treated GC cells. C Chromatin immunoprecipitation assay was conducted using extracts of MGC-803 transfected with ectopic HIF1A or control vector. IgG was used as negative controls (**p < 0.01) (left panel). Luciferase assay to show HIF1A regulating transcriptional activity of RhoA in MGC-803 cells (**p < 0.01) (right panel). D Western blotting to determine HIF1A, PHD3, ABHD16A, and RhoA protein levels in MGC-803 cells with ectopic miR-4646-5p, ectopic HIF1A or silenced PHD3 and the controls, or HIF1A reduced MGC-803 cells with ectopic miR-4646-5p or silenced PHD3. E, F Western blotting to detect p-LIMK and LIMK, p-cofilin and cofilin levels in the above GC cells. G–I Transwell assay to test cell invasion ability of GC cells mentioned in E and F. The histograms show the average invaded cells each view (**p < 0.01).
