Objective To explore the effect of overexpression of heparanase (HPSE) on the proliferation of gallbladder cancer cell line GBC-SD and the phosphatidylinositol 3-kinase/protein serine-threonine kinase (PI3K/AKT) signaling pathway.
Methods The HPSE eukaryotic expression vector was constructed, and then the gallbladder cancer cell line GBD-SD with transgenic overexpression of HPSE was obtained. The mRNA and protein expression of HPSE in GBC-SD and GBD-SD were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. The proliferation of cells in each group was assessed by colony formation assay and MTT. The protein levels of PI3K/AKT signaling pathway related proteins (p-PI3K, PI3K, p-AKT and AKT) were analyzed by western blot.
Results The mRNA and protein levels of HPSE in GBD-SD after successful transfection were significantly increased (P < 0.01). Compared to GBC-SD with normal expression of HPSE, the optical density (OD) value and colony formation ability of GBD-SD cells with overexpression of heparanase were significantly increased (P < 0.05). The results of western blot showed that compared with GBC-SD cells, the expression levels of p-AKT, p-PI3K protein, p-AKT/AKT value and p-PI3K/PI3K value in GBD-SD cells with overexpression of HPSE were increased significantly (P < 0.01), but the expression levels of total AKT and total PI3K protein did not change obviously. After 2 days of treatment with specific inhibitor LY294002 for PI3K/AKT pathway, the proliferation ability and p-AKT/AKT value of GBC-SD cells were significantly decreased (P < 0.01).
Conclusion Overexpression of HPSE can promote the proliferation of GBC-SD cells, which may be related to the PI3K/AKT signaling pathway.
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