ABSTRACT
Objective To investigate the effect of miR-200a on the migratory ability of NSCLC cells and to explore its possible mechanism.Methods Real-time PCR was performed to analyze the miR-200a expression in NSCLC cell lines A549 and SK-MES-1, and human normal lung bronchial epithelial cell line 16HBE.Hsa-miR-200a mimics, NC mimics, hsa-miR-200a inhibitor and NC inhibitor were transfected into A549 cells using Lipofectamine 2000.Migration of A549 cells was detected by Transwell migration assay.The potential target genes of miR-200a were predicted by bioinformatics software and then verified by dual luciferase reporter gene assay and Western blot.Results MiR-200a was significantly down-regulated in A549 and SK-MES-1 cells(P＜0.05).Exogenous over-expression of miR-200a mimics significantly inhibited migratory a-bility of A549 cells, while over-expression of miR-200a inhibitor generated the opposite effect(P＜0.01).Dual luciferase re-porter assay indicated that miR-200a could directly affect the 3'-UTR of Sulf2 gene to inhibit luciferase activity.Western blot revealed that miR-200a expression could significantly reduce Sulf2 protein expression level in A549 cells.Ectopic expression of Sulf2 protein in miR-200a-overexpressing A549 cells overrode the migration inhibition effect of miR-200a, suggesting that targe-ting Sulf2 represents an important mechanism of the anti-tumour activity of miR-200a in lung cancer.Conclusion MiR-200a inhibits migration of lung cancer cells by targeting Sulf2.
ABSTRACT
For decades, various systemic therapies have been explored for the treatment of advanced hepatocellular carcinoma (HCC), the fifth most common cancer and the third leading cause of cancer-related deaths worldwide. Nevertheless, no satisfactory results have been obtained so far. Glypican-3 (GPC-3) is a cell-surface heparan sulfate proteoglycans (HSPGs) that emerged as a promising diagnostic marker as well as target for therapy. Therefore; we investigated antitumor activity of antiglypican-3 (antiGPC3), a specific antibody aginst GPC-3, against HepG2, human HCC, cell line. HepG2 cells were treated with AntiGPC3 at (5, 10 and 20 μg/ml). HepG2 cell proliferation was measured by MTT and lactate dehydrogenase (LDH) assays. GPC-3, HSPG and sulfatase-2 (SULF2) levels were measured by ELISA. Moreover, apoptosis was assessed by measuring Caspase-3 activity. We found that, antiGPC3 reduced HepG2 cells survival and showed cell cytotoxicity in a dose-dependent manner. In addition, antiGPC3 was able to increase the apoptosis measured by capase-3 activity in hepG2 cells. Finally, antiGPC3 restored HSPG level without affecting SULF2 in HepG2 cells. We can conclude that, antiGPC3 possesses cytotoxic effects, which can be partially explained by restoration of HSPGs and increase of caspase-3 apoptotic pathway. GPC-3 represents a promising target of HCC therapy.