薏苡仁油对人结肠癌细胞增殖及凋亡的影响和机制

Effects of Coix Seed Oil on the Proliferation and Apoptosis of Human Colon Cancer Cells and Its Mechanism

目的:观察薏苡仁油对人结肠癌HT-29细胞增殖、凋亡的影响,探讨薏苡仁油对人结肠癌HT-29细胞Hedgehog(Hh)信号通路的影响。方法:分别使用不同浓度薏苡仁油处理人结肠癌细胞株HT-29,另取空白培养基做对照组。采用噻唑蓝还原法[3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]检测各组细胞增殖情况,采用流式细胞术检测各组细胞凋亡率。采用实时定量聚合酶链反应(quantitative polymerase chain reaction,q-PCR)检测Ptch、Smo、Shh和Gli-1 mRNA表达,采用蛋白印迹检测CyclinD1、p21、Caspase-3、Bcl-2、Ptch、Smo、Shh和Gli-1蛋白的表达。结果:MTT法观察到薏苡仁油可抑制人结肠癌HT-29细胞增殖,各浓度水平组HT-29细胞增殖抑制率差异均有统计学意义(P<0.05),抑制率随着薏苡仁油浓度的增加而升高。流式细胞术观察到薏苡仁油可诱导HT-29细胞凋亡,各浓度水平组HT-29细胞凋亡率差异均有统计学意义(P<0.05),凋亡率随着薏苡仁油浓度的增加而升高。与空白对照组比较,薏苡仁油各组Ptch、Smo、Shh和Gli-1 mRNA的表达降低,且随着薏苡仁油浓度增加,各组表达变化逐渐降低,差异有统计学意义(P<0.05)。与空白对照组比较,薏苡仁油各组CyclinD1、Bcl-2、Ptch、Smo、Shh和Gli-1蛋白的表达降低,且随着薏苡仁油浓度增加,各组表达变化逐渐降低,差异有统计学意义(P<0.05)。与空白对照组比较,薏苡仁油各组p21和Caspase-3蛋白的表达升高,且随着薏苡仁油浓度增加,各组表达变化逐渐升高,且差异有统计学意义(P<0.05)。结论:薏苡仁油能抑制人结肠癌HT-29细胞增殖、促进其凋亡,其机制可能和薏苡仁油抑制Hh信号通路相关。

Objective:To observe the effects of coix seed oil on the proliferation and the apoptosis of human colon cancer cells HT-29 cells,and explore the influence of coix seed oil on human colon cancer cells HT-29 cells Hedgehog(Hh)signaling pathway.Methods:After dealing human colon cancer cells HT-29 cells with different concentrations of coix seed oil,blank medium was chosen as the control group,MTT method was adopted to detect the proliferations of the cells in different groups,flow cytometry was used to measure the apoptosis rate of the cells in different groups.q-PCR was used to detect the expressions of Ptch,Smo,Shh and Gli-1 mRNA,Western blotting was used to measure the expressions of Cyclin D1,p21,Caspase-3,Bcl-2,Ptch,Smo,Shh and Gli-1 proteins.Results:MTT assay showed that coix seed oil could inhibit the proliferation of human colon cancer cells HT-29 cells,the difference had statistical meaning in the inhibitory rate of HT-29 cells proliferation between different concentrations groups(P<0.05),the inhibitory rate increased as the concentrations of coix seed oil raised.Flow cytometry displayed that coix seed oil could induce the apoptosis of HT-29 cells,the difference had statistical meaning in the apoptosis rate of HT-29 cells between different concentrations groups(P<0.05),the apoptosis rate rose as the concentrations of coix seed oil increased.Compared with blank control group,the expressions of Ptch,Smo,Shh and Gli-1 mRNA in different groups of coix seed oil reduced,the expressions of different groups lowered as the concentrations of coix seed oil increased,and the difference was statistically significant(P<0.05).Compared with blank control group,the expressions of Cyclin D1,Bcl-2,Ptch,Smo,Shh and Gli-1 proteins reduced in different groups of coix seed oil,the expressions gradually reduced in different groups as the concentrations of coix seed oil raised,and the difference showed statistical meaning(P<0.05).Compared with blank control group,the expressions of p21 and Caspase-3 protein raised in different groups of coix seed oil,the expressions rose gradually in different groups as the concentrations of coix seed oil raised,and the difference showed statistical meaning(P<0.05).Conclusion:Coix seed oil could inhibit the proliferation of human colon cancer cells HT-29 cells and promote its apopotsis,and its mechanism migth be related to that coix seed oil could inhibit Hh signaling pathway.