DPC4基因转染对胰腺癌JF305细胞凋亡的影响

摘要

目的:探讨DPC4基因对人胰腺癌JF305细胞凋亡的影响.方法:采用脂质体介导法将pBK-CMV-DPC4质粒导入JF305中,以转染空质粒pBK-CMV和未转染的JF305为对照.采用流式细胞仪法检测3种细胞中Bcl-2和Bax蛋白的表达,MTT法检测细胞增殖率,流式细胞仪法检测细胞周期,免疫印迹法检测DPC4蛋白的表达.结果:与未转染及转染空质粒组的细胞比较,转染pBK-CMV-DPC4的JF305细胞增殖率降低,细胞周期阻滞在G1期(F=41 722.447,P<0.001),DPC4蛋白表达增强(F=10 821.545,P<0.001),Bcl-2蛋白表达降低(F=387.173,P<0.001),Bax蛋白表达增强(F=250.808,P<0.001),Bcl-2/Bax的比值降低(F=776.568,P<0.001).结论:DPC4蛋白通过调节Bcl-2和Bax的表达,促进胰腺癌细胞凋亡.%Aim: To explore the effect of DPC4 gene on the cell apoptosis of human pancreatic carcinom JF305 cells. Methods;Human pancreatic adenocarcinoma cell line JF3O5 was transfected with pBK-CMV-DPC4 plasmid using lipo-fectamine transfection technique, and cells transfected with pBK-CMV plasmid or without transfection were the controls. The expression of DPC4 protein were studied by Western blot. The cell growth was estimated by MTT method. The expressions of Bcl-2 and Bax proteins were detected by flow cytometry. Results: Compared with those of the controls, the proliferation of the cells transfected with pBK-CMV-DPC4 was suppressed significantly, and maintained in Gl phase(F=41 722.447, P < 0. 001) , the expression of DPC4 protein was higher ( F = 10 821. 545 , P < 0. 001) , the Bcl-2 expression was lower ( F = 387. 173,P<0. 001) ,the Bax expression was higher( F = 250. 808 ,P <0. 001) ,and Bcl-2/Bax was lower( F = 776. 568 , P < 0.001). Conclusion: DPC4 could induce JF305 cell apoptosis possibly through regulating the expression of Bcl-2 and Bax proteins.