摘要
目的:检测大豆异黄酮代谢产物雌马酚对大肠癌细胞增殖的影响,并探讨其分子作用机制。方法:培养大肠癌细胞DLD1、HCT15、COLO205、LOVO和SW480,用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]法检测雌马酚对大肠癌细胞增殖的影响,用逆转录PCR和蛋白免疫印迹法分别检测雌激素受体和核因子红系2相关因子2[nuclear factor(erythroid-derived 2)-like 2,Nrf2]表达状况。用MTT法检测雌激素受体抑制剂和激动剂对大肠癌细胞增殖的影响。结果:DLD1、HCT15、COLO205、LOVO和SW480均表达雌激素受体(estrogen receptor,ER)β,但只有DLD1和HCT15有ERα弱表达。MTT试验显示不同剂量雌马酚(0、0.5、1、5、10μmol/L)不仅能够显著抑制ERα和ERβ均表达的大肠癌细胞HCT-15的增生,且呈现明显的剂量效应关系,而且能够抑制只有ERβ表达的大肠癌细胞LOVO和SW480的增生,且呈现明显的剂量效应关系。此外,逆转录PCR发现不同剂量雌马酚刺激后,HCT-15的雌激素受体ERα和ERβ表达明显增加,呈剂量效应关系。蛋白免疫印迹法进一步证实雌激素受体ERα和ERβ蛋白表达随雌马酚剂量的增加而增加,蛋白免疫印迹法发现Nrf2随干预剂量的增加而表达明显增加。采用ERβ激动剂、雌激素受体抑制剂和特异性雌激素受体ERα激动剂对SW480、LOVO、HCT-15细胞刺激后,只发现不同剂量ERβ激动剂能够显著抑制大肠癌细胞SW480、LOVO、HCT-15的增生,且呈明显的剂量效应关系,但是,雌激素受体抑制剂和特异性雌激素受体ERα激动剂对SW480、LOVO和HCT-15细胞没有呈现显著的影响。结论:雌马酚能够通过雌激素样作用和抗氧化活性抑制大肠癌细胞的增殖。
Objective: To investigate the effect of equol on the proliferation of colom cancer cells and to explore the mechanisms. Methods: Colon cancer cells( DLD1,HCT15,COLO205,LOVO,SW480) were incubated,the cell proliferation was identified by 3-( 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay. Reverse transcription PCR and Western blot were used to measure the mRNA and the protein expression of estrogen receptor and nuclear factor( erythroid-derived 2)-like 2( Nrf2) in the colon cancer cells,respectively. Moreover,3-( 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay was used to investigate the effect of estrogen receptor( ER) inhibitor,ERα agonist,and estrogen receptor ERβagonist on the cell proliferation. Results: ERα was faintly expressed in the DLD-1 and HCT-15 cells. However,ERβ expression in DLD1,HCT15,COLO205,LOVO,and SW480 colon cancer cells. Different concentrations of equol( 0,0. 5,1,5,10μmol/L) significantly inhibited the growth of HCT-15 cell with the expression of ERα and ERβ. Moreover,different concentrations of equol( 0,0. 5,1,5,10 μmol/L) significantly inhibited the growth of LOVO,and SW480 cells with the ERβ expression in a dose-dependent manner as demonstrated with a 3-( 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay. mRNA expressions of ERα and ERβ in HCT-15 were stimulated significantly. Western blotting proved that the protein expressions of ERα and ERβ increased with the increasing of equol dose. Moreover we found significant difference of Nrf2 protein expression in HCT-15 cell stimulated by different concentrationss of equol. After the similation of estrogen receptor inhibitor,ERα agonist,or ERβ agonist,we found that only different concentrations of ERβ agonist( 0,1,10,100,1 000,10 000 nmol/L) significantly inhibited the growth of HCT-15,LOVO,and SW480 in adose-dependent manner. Estrogen receptor inhibitor and ERαagonistdid not present significant effect on the cell proliferati
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2017年第3期383-387,共5页
Journal of Peking University:Health Sciences
基金
国家自然科学基金(81202193
81573130)
北京市自然科学基金(7122103
7172117)资助~~
关键词
雌马酚
大肠癌
雌激素受体
增殖
Equol
Colon cancer
Estrogen receptor
Proliferation
