The Effect Of Low-expression PDCL3 On VEGF In SKOV3 Cell Line

Objective:Ovarian cancer poses a serious threat for women.Recently,antiangiogenic therapy based on inhibiting VEGF/NEGFR2 signaling pathway improved the clinical outcome,however the regulation mechanism of VEGF/VEGFR2 signaling pathway in ovarian cancer is unclear.Previous studies identified PDCL3 as a novel chaperone protein of VEGFR2,controling the abundance of VEGFR-2 and angiogenesis positively.In this study,the effect of PDCL3 on VEGF in SKOV3 cell line was explored,so was the antiangiogenic therapy-Bevacizumab-on PDCL3.Methods:1.PDCL3-SiRNA was transfected into SKOV3 cell line by lentivirus for 72h to silence PDCL3.Efficiency of transfection was observed by fluorescence microscope.2.The integrity of total RNA was examined by agarose gel electrophoresis.3.qPCR and Western blot were used to identify the down-regulation of PDCL3 and examine the relative quantity of mRNA and protein relative expression of VEGF,VEGFR2 respectively,p-AKT and p-ERK were examined by Western blot.4.Different concentrations of Bevacizumab was used to treat SKOV3 cells for 24h:0(control group),50,100,150mg/L.The relative quantity of mRNA and protein expression for PDCL3 were examined by qPCR and Western blotResults:1.Fluorescence microscope showed the efficiency of transfection was above 80%.2.The bands of 5s,18s,28s rRNA was clear and could be found on the image system of agarose gel electrophoresis.3.qPCR and Western blot confirmed that the mRNA and protein expression of PDCL3 were both significantly decreased(P<0.05);the mRNA expression of VEGFR2 did not change but its protein expression is decreased in PDCL3-SiRNA group(P<0.05);no change was observed in VEGF,p-AKT and p-ERK(P>0.05).4.qPCR and Western blot confirmed that the mRNA and protein expression of PDCL3 in Bevacizumab group(50,100,150mg/L)was significantly increased SKOV3 cells(P<0.05).Conclusion:1.After PDCL3 is down-regulated,the protein expression of VEGFR2 is significantly decreased,PDCL3 might regulate have a regulatory role on VEGFR2 in SKOV3 cells.2.Down-regulation of PDCL3 has no effect on the expreession of VEGF,p-AKT,p-ERK.3.Bevacizumab of certain concentrations enchances the mRNA and protein expression of PDCL3 in SKOV3 cells.