大萼香茶菜甲素对多发性骨髓瘤细胞株U266体外作用的研究(1)
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摘 要：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；Annexin V+ /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。
关键词：大萼香茶菜甲素；U266细胞；增殖；细胞凋亡
中图分类号：R285.5
文献标识码：A
文章编号：1673-7717(2012)04-0723-04
Effect of macrocalin A on myeloma U 266 Cells in Vitro
FENG Li-qian1,2,ZHOU Yong-lie2，LU Ya-ping3, XIA Jun2, QIU Lian-nv2, SHI Hao3
(1.Zhejiang Provincial People’s Hospital,Hangzhou 310014，Zhejiang，China;
2.Zhejiang Chinese MedicalUniversity,Hangzhou 310054，Zhejiang，China;
3.The Medical Colleage of Zhejiang Industry University,Hangzhou 310014，Zhejiang，China)
Abstract:Objective:To investigate the effect of macrocalin A (MA) inducing apoptosis on myeloma U266 cell in vitro. Methods: In vitro,U266 cells were incubated with different concentrations (2、4、8μg/mL)of MA, the cells’growth was tested by cell counting, MTT assay. Cell morphology，Annexin - V/PI stainning， DNA subdiploid content, were used to investigate the apoptosis of U266. The expression of proteinβ1、β1i、β2、β2i、β5、β5i、NF-κB、IκB-αwas detected by Western blot technique.Results: Different concentrations ofMA could obviously inhibit the growth of U266 cells with time and dose dependence. Characteristic apoptostic body was showed,annexinV - F ITC + cell and significant increase of sub - G1 cells showed MA induce apoptosis.Western blot analysis showed that the protern expression ofβ2、β1i、β5i、NF-κB was down-regulated in U266 cells treared with MA but the protern expression of IκB was advanced. Conclusion: MA can inhibit proliferation and induced apoptosis of U266 cell.
Key words:Macrocalin A；myeloma U266 cell;; Proliferation; Apoptosis
大萼香茶菜Rabdosia macrocalyx(Dunn)Hara系唇形科香茶属植物，在国内华东、华南广泛分布，具有清热、消炎等作用，长期以来约有10余种香茶属植物在民间作为草药广泛应用。近年来，发现其中的二萜类化合物具有抗肿瘤的活性，其中的α亚甲基环戊酮基团被认为是抗肿瘤的活性基团。与此相似的如紫草素、冬凌草甲素、毛萼乙素等都具有潜在的抗肿瘤活性[1-3]。大萼香茶菜甲素(Macrocalyxin A, MA)是从大萼香茶叶醇提取物中分离结晶而得 ......
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