Overexpression Of Heparanase Affects The Proliferation Of Gallbladder Carcinoma Cells By Regulating PI3K/AKT Signaling Pathway

Purpose The purpose of this study was to investigate the effect of overexpressed Heparanase on the proliferation and PI3K/AKT signaling pathway of gallbladder cancer cell line GBC-SD.Methods 1.The Heparanase eukaryotic expression vector was constructed,and then the gallbladder cancer cell line GBD-SD with transgenic overexpression of Heparanase was obtained.2.In order to study the expression of Heparanase in gallbladder cancer cell lines GBC-SD and GBD-SD,we compared and analyzed the m RNA and protein expression of Heparanase in gallbladder cancer cell lines GBC-SD and GBD-SD by q RT-PCR and Western blot experiments.3.The proliferation of GBC-SD and GBD-SD cells was detected by MTT method and colony formation assay.4.The effect of Heparanase overexpression on the expression of PI3K/AKT signaling pathway-related proteins（p-PI3 K,PI3K,p-AKT and AKT）was detected by Western blot assay.5.To further explore whether Heparanase promotes the proliferation of GBC-SD cells through the PI3K/AKT signaling pathway,in this study,GBC-SD cells were treated with PI3K/AKT signaling pathway inhibitor LY294002（10umol/L）for 2 days,and the proliferation of GBC-SD group and LY294002 group was detected by MTT method and colony formation assay.Results 1.The relative expressions of Heparanase m RNA and protein in GBD-SD cells after successful transfection were higher than those in GBC-SD cells,and the difference was statistically significant（P<0.01）.2.Compared to GBC-SD with normal expression of Heparanase,the optical density（OD）value and colony formation ability of GBD-SD cells with overexpression of heparanase were significantly increased,and the difference was statistically significant（P<0.05）.3.Western blotting results showed that compared with the control group GBC-SD cells,the protein expressions of p-AKT and p-PI3 K in the GBD-SD group cells were significantly increased,while the levels of total AKT and total PI3 K did not change significantly,but the values of p-AKT/AKT and p-PI3K/PI3 K were significantly increased,and the difference was statistically significant（P<0.01）.4.Compared with control group GBC-SD cells,the proliferation ability and p-AKT/AKT value of LY294002 group cells were significantly decreased,and the difference was statistically significant（P<0.01）.Conclusion Overexpression of heparanase can promote the proliferation of GBC-SD cells,and this effect may be related to the PI3K/AKT signaling pathway.