白藜芦醇抑制人结肠癌细胞增殖的作用与促进骨形态蛋白9及其受体表达的关系研究

摘要

Aim To study the anti-proliferation effect of resveratrol (Res)and the role of Res-induced bone morphorgenetic protein 9 (BMP9 )in this process in colon cancer cells.Methods Crystal violet staining and flow cytomtry were introduced to assay the anti-proliferation effect of Res in LoVo cells.The effect of Res on apoptosis in LoVo cells was also detected with flow cytometry.Then,RT-PCR and Western blot assay were employed to unveil the effect of Res on the ex-pression of BMP9 .The effect of BMP9 on the anti-pro-liferation of Res in LoVo cells was analyzed with crystal violet staining and flow cytometry too.Finally,the effect of Res on the expression of ALK2 and ALK3 was assayed with RT-PCR,and the inhibitor of ALK2 and ALK3 was used to figure out the possible mechanism of BMP9 on Res-induced proliferation inhibition in LoVo cells.Results Res apparently inhibited the prolifera-tion,arrested the cell cycle at S phase in LoVo and in-creased the percentage of apopotic cells in LoVo cells. Res increased the expression of mRNA and protein of BMP9 concentration dependently. Exogenous ex-pressed-BMP9 enhanced the anti-proliferation and ap-optosis inducing effects of Res in LoVo cells, but BMP9 knockdown decreased these effects of Res.Al-though Res had no apparent effect on increasing the phosphorylation of Smad1/5/8,it increased the ex-pression of ALK2 and ALK3 .Inhibition of ALK2 and ALK3 decreased the anti-proliferation effect of Res partly in LoVo cells.Conclusion Res is potent to in-hibit the proliferation of LoVo cells,Which may be mediated by up-regulating the expression of BMP9 and its receptor at least.%目的：研究白藜芦醇（resveratrol，Res）对人结肠癌Lo-Vo细胞增殖的抑制作用与骨形态蛋白9（bone morphorgenet-ic protein 9，BMP9）之间的可能关系。方法采用结晶紫染色与流式细胞术检测Res对LoVo细胞增殖的抑制作用；流式细胞术检测Res对LoVo细胞凋亡的诱导作用。利用RT-PCR和Western blot检测Res在LoVo细胞中对BMP9表达的影响，以及对 BMP9的受体 ALK2和 ALK3表达的影响。采用ALK2和ALK3的抑制剂，通过结晶紫染色分析BMP9介导Res抑制LoVo细胞增殖的可能机制。结果与对照组相比，Res明显呈浓度依赖性抑制 LoVo 细胞增殖，并诱导LoVo细胞发生S期阻滞；Res呈浓度依赖性增加LoVo细胞凋亡比例；Res促进BMP9的mRNA表达并明显增加其蛋白水平；外源性过表达BMP9能增强Res对LoVo细胞的增殖抑制和诱导凋亡作用；Res 能诱导 BMP9的受体 ALK2和ALK3的mRNA表达，抑制ALK2和ALK3能明显促进LoVo细胞增殖，并减弱Res对LoVo细胞增殖的抑制作用。结论Res 对LoVo 细胞的增殖具有明显抑制作用，这种作用可能与Res 促进BMP9及其受体表达有关。