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目的体外观察组蛋白去乙酰化酶抑制剂TSA、蛋白酶体抑制剂MG-132单独和联合作用致骨髓瘤及淋巴瘤细胞增殖阻滞及凋亡。方法TSA和MG-132以不同浓度单独和联合作用细胞株EL4、WEHI-231、SP2/0、P3NSI,台盼蓝法、AnnexinV/propidium iodide染色、流式细胞仪观察细胞活力、周期和凋亡。Bliss法计算TSA、MG-132对各细胞株的IC_(50)值,Calcuysn软件评价药物协同效应。免疫印迹检测凋亡、周期相关蛋白Bcl-2、Bax、caspase3、CyclinD1、p21的表达,观察转录因子IRF4和c-Myc表达水平。结果TSA和MG-132单独作用均可呈药物浓度依赖性地抑制细胞增殖、诱导细胞凋亡,两药协同效应显著。细胞凋亡相关蛋白caspase-3激活,Bax、Bcl-2表达均相对下调,阻滞于G_0～G_1期的细胞明显增多,CyclinD1表达水平下降,p21水平显著上升。结论TSA和MG-132单独作用细胞株WEHI-231、EL4、SP2/0、P3NSI,呈药物浓度依赖性地抑制细胞增殖、激活caspase-3诱导细胞凋亡,两药合用呈显著协同效应。同时,两药单独或联合作用均能致细胞G_1期阻滞。
Objective To observe the proliferation and apoptosis of myeloma and lymphoma cells induced by histone deacetylase inhibitor TSA and proteasome inhibitor MG-132 in vitro and in vivo. Methods TSA and MG-132 cells were stained with EL4, WEHI-231, SP2 / 0, P3NSI, Annexin V / propidium iodide and Annexin V / propidium iodide at different concentrations respectively. Cell viability, . Bliss method to calculate TSA, MG-132 on each cell line IC 50 values, Calcuysn software evaluation of drug synergies. The expression of Bcl-2, Bax, caspase3, CyclinD1, p21were detected by Western blot, and the expression of IRF4and c-Myc were observed. Results TSA and MG-132 alone could inhibit cell proliferation and induce apoptosis in a concentration-dependent manner. The synergistic effect of both drugs was significant. The activation of caspase-3, the expression of Bax and Bcl-2 were all down-regulated. The number of cells arrested in G_0 ~ G_1 phase increased significantly, the expression of CyclinD1 decreased and the level of p21 increased significantly. Conclusion TSA and MG-132 alone act as cell lines WEHI-231, EL4, SP2 / 0 and P3NSI in a concentration-dependent manner to inhibit cell proliferation and activate caspase-3 to induce apoptosis. At the same time, the two drugs alone or in combination can cause G_1 cell arrest.