芒柄花黄素对结直肠癌细胞增殖、凋亡的影响及机制探讨

摘要

目的 观察芒柄花黄素对结直肠癌细胞增殖、凋亡的影响,并探讨其机制.方法 将结直肠癌细胞HCT116随机分为两组,观察组分别以25、50、100μmol/L芒柄花黄素处理,对照组以等量二甲基亚砜处理;用MTT法检测细胞增殖情况(A490),Hoechst荧光染色法计数凋亡细胞,qRT-PCR法检测细胞中的HOC转录反义RNA(HOTAIR),Western blot法检测细胞中的Bax和Bcl-2.结果 与对照组比较,观察组随着芒柄花黄素浓度增加及处理时间延长HCT116细胞A490值降低(P均<0.05);观察组以25、50、100μmol/L芒柄花黄素处理48 h后HCT116细胞凋亡数分别为(3.20±0.84)、(20.20±1.92)、(31.00±2.92)个/HP,均高于对照组的(1.6±1.14)个/HP(P均<0.01);与对照组比较,观察组细胞HOTAIR、Bcl-2表达量降低,而Bax表达量增加,P均<0.05.结论 芒柄花黄素可能通过抑制HOTAIR调节Bax和Bcl-2的表达,从而抑制结直肠癌细胞的生长并诱导细胞凋亡.%Objective To observe the effects of formononetin on the proliferation and apoptosis of colorectal cancer cells in vitro and to study the underlying mechanism.Methods The colorectal cancer cells HCT116 were randomly divid-ed into two groups:the observation group and the control group.Cells in the observation group were treated with 25μmol/L, 50 μmol/L and 100 μmol/L formononetin, and the control group was treated with the same amount of dimethyl sulfox-ide.The proliferation ( A490 ) of HCT116 cells measured by MTT assay, the apoptosis was assessed with Hoechst 33258 staining, the expression level of HOC transcription antisense RNA ( HOTAIR) was quantified by qRT-PCR, and the ex-pression level of Bax protein and Bcl-2 protein was determined by Western blotting.Results Compared with the control group, the A490 values of the HCT116 cells constantly decreased along with the increased doses and prolonged time in the observation group (all P<0.05).The numbers of apoptosis cells in the observation group treated with 25 μmol/L, 50μmol/L and 100 μmol/L formononetin after 48 h were respectively (3.20 ±0.84), (20.20 ±1.92) and (31.00 ± 2.92)/HP, which were all higher than that[(1.6 ±1.14)/HP]of the control group (all P<0.01).Compared with the control group, the expression of Bcl-2 protein and HOTAIR was significantly decreased, but the expression of Bcl-2 protein was significantly increased in the observation group (all P<0.05).Conclusion Formononetin may inhibit the growth and induce the apoptosis of colorectal cancer cells by inhibiting HOTAIR to regulate the expression of Bcl-2 and Bax protein.