八棱丝瓜蛋白-1对B16细胞的诱导分化作用

Differentiation-inducing effect of Luffaculin-1 on B16 melanoma cells

目的:测定八棱丝瓜蛋白-1对小鼠黑色素瘤B16细胞的诱导分化作用.方法:以MTT 法测定八棱丝瓜蛋白-1 对B16细胞的增殖抑制作用,以形态、黑色素含量及细胞周期变化为指标,观察八棱丝瓜蛋白-1诱导B16细胞分化的作用.结果:八棱丝瓜蛋白-1 对B16 细胞增殖具有明显的抑制作用,呈剂量依赖性和时间依赖性,LF-1对B16细胞作用48和72 h的IC50分别为33.29和10.28 g/L,并阻止B16细胞由G1期向S期过渡从而停滞于G1期;细胞形态向正常上皮样细胞分化,生长缓慢,平铺不重叠,甚至形成网状结构,胞质内细胞器丰富,可见较多不同成熟期(Ⅰ～Ⅳ期)的黑色素小体;黑色素含量显著增加,2.5、5、10和20 mg/L浓度的LF-1处理组的黑色素含量分别为对照组的2.1、4.4、5.9和7.8倍.结论:八棱丝瓜蛋白-1对B16细胞具有明显的诱导分化作用.

OBJECTIVE： To investigate the differentiation-inducing effect of Luffaeulin-1 on B16 mouse melanoma cells. METHODS：Evaluate the inhibitory effect of Luffaeulin-1 on proliferation of B16 cells by MTT assay. Estimate the differentiation-inducing activity of Luffaculin 1 in B16 cells, by observations on morphology, melanin content and dislrihution of cell cycle. RESULTS： Luffaeulin-1 significantly inhibited the proliferation of B16 cells in dose- and time-dependent manners, and IC50, of 48 and 72 h were 33.29 and 10. 28 g/L, respectively. Luffaculin-1 interrupted the transition of cell cycle from G1 phase to S phase, and arrested at the G1 phase of cell cycle. With Luffaculin-1 treatment, B16 cells were induced to differentiate into mature epithella-like cells, grew slowly in monolayer and even formed reticulation. Luffaeulin-1 also en riched cell organelles in cytoplasm, and melanosomes of different stages （Ⅰ-Ⅳ） can be observed. Meanwhile the content of melanin enhanced markedly. Compared with control, melanin contents in 2.5, 5, 10 and 20 mg/L of Luffaeulin-1 treated groups were 2.1, 4.4, 5.9 and 7.8 times, respectively. CONCLUSION： Luffaeulin-1 can significantly induce differentiation of B16 cells.