The Empirical Study Of NF-KB P65 ASODN Induces Appoptosis In Human Gastric Cancer Cells SGC-7901

Objective:The investigation used NF-κB p65 ASODN technique to block objective genes specificity, approached the the effect of NF-κB p65 ASODN on proliferation and apoptosis and change in form of SGC-7901, invested the effect of apotosis in SGC-7901 by down regulatiing the expression of NF-κB p65, approached the Mechanism of action of inducing apotosis.Methods:Man-made synthesis NF-κBp65 ASODN, Transfecting SGC-7901 trough liposome infection protocol. The experiment can be divided into groups: control group, Lip group, ASODN+Lip group, MSODN+Lip group. ASODN was divided with different concentrations (10μM,20μM,40μM) and collecting cells in different times(24h,48h,72h). The effect of NF-κB p65 ASODN on cell proliferation was observed by MTT colormetric assay. The rate of SGC-7901apoptosis and cell cycle was identified by flow cytometry (FCM) and the morphological change of apoptosis was observed by light microscopy. And then the expression of apoptosis-regulating protein NF-κB p65, Bcl-2 in SGC-7901 after apoptosis induced by NF-κBp65 ASODN were examined by immunocytochemical staining assay .Result:①MTT analysis: NF-κB p65 ASODN with different concentrations incubated with SGC-7901 for 24h, 48h and 72h could significantly inhibit SGC-7901 proliferation in dose-dependent and time-dependent manner compared with the control group(p<0.05);②Flow cytometry quantitation: NF-κB p65 ASODN could induce cells apoptosis and block cells in G0/G1 time. The apoptosis ratio of ASODN+Lip group was (50.51±3.35)%, which was significantly higher than that of MSODN+Lip group (20.34±2.11)% (P<0.05).③HE staining: Treatment with 40μM concentrations of Ursolic acid for 48h resulted in morphologic changes of SGC-7901, including karyorrhexis and cytoplasm vacuolization.④Immunocytochemistry: NF-κB p65 ASODN incubated with SGC-7901 for 48h, the expressions of NF-κB p65, bcl-2 protein in SGC-7901 were significantly up-regulated(P<0.05).Conclusion:①The compounds with NF-κB p65 ASODN and liposome could transfected into SGC-7901 successfully.②NF-κB p65 ASODN could significantly inhibit SGC-7901 proliferation and induce apoptosis in dose-dependent and time-dependent manner.③NF-κB p65 ASODN could down regulated the expressions of NF-κB p65 and Bcl-2 protein in SGC-7901.④Our results suggested that SGC-7901 cells apoptosis induced by NF-κB p65 ASODN through mechanisms was that it could transfected into SGC-7901 with liposome, then specific bind with NF-κB p65 mRNA, inhibit and block the expression of NF-κB p65 mRNA, reduce the expressions of NF-κB p65 and Bcl-2 protein, thus inhibit proliferation and induce cells apoptosis at last.