和厚朴酚抑制结肠癌细胞增殖与骨形态发生蛋白7关系的研究

Inhibitory growth effect of honokiol and BMP7 in human colon cancer cells

目的研究和厚朴酚(honokial,HNK)对人源结肠癌细胞HCT116增殖的影响及其可能分子机制。方法采用结晶紫染色、流式细胞术检测和厚朴酚对HCT116细胞增殖和细胞周期的影响;Western blot分析和厚朴酚对增殖细胞核抗原(PCNA)及细胞凋亡指标Bcl-2的影响;Western blot及半定量PCR分析和厚朴酚对HCT116细胞中内源性骨形态蛋白7(BMP7)表达的影响;利用BMP7过表达腺病毒及抗体,通过结晶紫定量和Western blot,分析和厚朴酚联合应用BMP7腺病毒与抗体对HCT116细胞增殖的影响。结果和厚朴酚呈浓度和时间依赖性地抑制HCT116细胞增殖、诱导细胞凋亡、使细胞阻滞于G_2期,并上调PCNA的蛋白表达水平,下调Bcl-2的蛋白表达水平;和厚朴酚可上调HCT116细胞中BMP7的mRNA及蛋白表达水平;外源性BMP7过表达腺病毒可促进和厚朴酚的增殖抑制和促凋亡作用,而BMP7抗体可抑制此作用。结论和厚朴酚能够抑制HCT116的增殖并促进凋亡,其机制可能与上调BMP7的表达有关。

Aim To investigate the anti-proliferation effect of honokial on human colon cancer cells HCT116 and the possible mechanism.Methods Crystal staining and flow cytometry assay were introduced to test the proliferation inhibitory and cell cycle arrested effect of honokial on HCT116 cells.Western blot was used to analyse the expression of PCNA and induction effect of apoptosis.Western blot and PCR assay were conducted to assess the change of BMP7.Via exogenous adenoviruses of BMP7 and its antibody combined with honokial,crystal violet staining and Western blot were used to analyse the effect on HCT116 cells.Results Honokial inhibited the growth of HCT116 in a time-and concentration-dependent way,induced apoptosis and arrested at G_2 phase; Western blot assay showed honokial up-regulated the level of PCNA and BMP7,and downregulated the expression of Bcl-2; Western blot result also showed that exogenous adenoviruses of BMP7 enhanced the effect of honokial on proliferation inhibition and apoptosis,while BMP7 antibody reversed such effects of honokial.Conclusion Honokial can inhibit the proliferation and induce apoptosis on HCT116 cells,which may be mediated by the up-regulation of BMP7.