Study On The Antitumor Activity And Mechanism Of Mapat

ObjectiveTo investigate the anti-tumor activity and mechanism of a amidothiourea acridine derivate MAPAT in vitro.Methods1. The inhibitory effects of MAPAT on CNE-2, A549, panc-1, SN-K and Hela cells proliferation were analyzed by CCK-8 assay.2. Morphology of cell was investigated by HE staining.3.Cellular ultrastructure was observed by transmission electron microscope.4. The effect of MAPAT on apoptotic rate and cell cycle distribution were evaluated by flow cytometry (FCM).5. The mRNA expression levels of Topo I, Topo II a, Topo Ⅱβ were detected by real-time PCR.6. The protein expressions of Topo I, Topo II a, Topo Ⅱβ were assessed by Western blot.Results1. CCK-8 assay revealed that the MAPAT could inhibit the proliferation of CNE-2, panc-1, SN-K, A549 and Hela cells. The IC50 values were (8.90±0.49),(44.44±0.02),(133.54±0.87),(80.16±0.18),(92.77±0.12)μmol/L repectively.2.The results of flow cytometry indicated that 48h after treatment,the apoptosis ratios were (2.63±0.21)%, (2.70±1.19)%, (38.80±0.1)%, respectively.MAPAT significantly arrested cell cycle a S phase in A549 cell.3.RT-PCR and western blot showed that MAPAT could decrease the mRNA and protein expression of Topo II a,while increased the levels of Topo I and Topo Ⅱβ.ConclusionMAPAT could inhibit CNE-2, panc-1, SN-K, A549 and Hela cells proliferation.the mechanism might be associated with the induction of tumor cell apoptosis and the up-regulation of Topo I and Topo Ⅱβ.