目的研究白藜芦醇(resveratrol,Res)对人结肠癌LoVo细胞增殖的抑制作用与骨形态蛋白9(bone morphorgenetic 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RTPCR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。
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 antiproliferation 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 assaywere employed to unveil the effect of Res on the expression of BMP9. The effect of BMP9 on the anti-proliferation 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 LoVocells. Results Res apparently inhibited the proliferation,arrested the cell cycle at S phase in LoVo and increased the percentage of apopotic cells in LoVo cells.Res increased the expression of mRNA and protein of BMP9 concentration dependently. Exogenous expressed-BMP9 enhanced the anti-proliferation and apoptosis inducing effects of Res in LoVo cells, but BMP9 knockdown decreased these effects of Res. Although Res had no apparent effect on increasing thephosphorylation of Smad1 /5 /8,it increased the expression 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 inhibit the proliferation of LoVo cells,Which may be mediated by up-regulating the expression of BMP9 and its receptor at least.