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目的:观察大萼香茶菜甲素(macrocalyxin A,MA)体外对多发性骨髓瘤细胞株U266增殖、凋亡的作用,并初步探讨其机制。方法:以不同质量浓度的(2、4、8μg/mL)MA体外作用于U266细胞,运用细胞计数、PI染色、MTT比色法体外观察MA对U266细胞增殖抑制作用;采用细胞形态观察、Annexin V/PI双染色和DNA亚二倍体检测方法研究MA诱导U266细胞凋亡作用;用Western免疫印迹法检测β1、β1i、β2、β2i、β5、β5i、NF-κB、IκB-α蛋白表达。结果:MA抑制细胞增殖;MA可促进细胞凋亡,细胞瑞氏染色后出现典型的凋亡小体;AnnexinV+/P I-表达升高;亚G1峰显著增加;在Western免疫印迹中β2、β1i、β5i、NF-κB随着药物作用浓度的增高表达逐渐降低,而IκB为增高趋势,β1、β2、β2i无明显变化。结论:MA在体外对U266细胞增殖有明显抑制作用,并诱导细胞调亡。
Objective: To observe the effect of macrocyrotic A (MA) on the proliferation and apoptosis of multiple myeloma cell line U266 in vitro and to explore its mechanism. Methods: U266 cells were treated with different concentrations of MA (2, 4, 8μg / mL) in vitro. The proliferation of U266 cells was observed by MTT colorimetric assay, cell counting, PI staining and MTT assay. V / PI double staining and DNA sub-diploid assay were used to study the apoptosis of U266 cells induced by MA. The protein expressions of β1, β1i, β2, β2i, β5, β5i, NF-κB and IκB-α were detected by Western blotting. Results: MA inhibited cell proliferation; MA promoted cell apoptosis; typical apoptotic bodies appeared after Wright’s staining; the expression of Annexin V + / P I- increased; the sub-G1 peak increased significantly; in Western immunoblots, β2, , Β5i and NF-κB gradually decreased with the increase of drug concentration, while IκB increased and β1, β2 and β2i did not change significantly. Conclusion: MA can obviously inhibit the proliferation of U266 cells in vitro and induce cell apoptosis.