Inhibitory effects of cholesterol sulfate on progesterone production in human granulosa-like tumor cell line, KGN.
Inhibitory effects of cholesterol sulfate on progesterone production in human granulosa-like tumor cell line, KGN.
复制标题
硫酸胆固醇对人颗粒样肿瘤细胞系 KGN 中黄体酮产生的抑制作用。
作者：
R. Tsutsumi;H. Hiroi;M. Momoeda;Yumi Hosokawa;F. Nakazawa;M. Koizumi;T. Yano;O. Tsutsumi;Y. Taketani
硫酸胆固醇（CS）是细胞膜的成分，具有稳定细胞膜的作用。我们之前报道过，CS在着床期间在子宫内膜中增加，这表明CS在生殖中发挥着重要作用。据报道，CS 调节胎盘、肾上腺和卵巢中黄体酮和孕烯醇酮的产生。然而，CS 产生类固醇激素的调节机制仍然未知。在本研究中，我们研究了CS对源自人颗粒样肿瘤的KGN细胞中黄体酮产生相关基因表达的影响。 KGN 细胞在 8-溴-cAMP (1 mM) 存在下与 CS (10 muM) 或胆固醇 (10 muM) 一起培养。 CS和cAMP处理后24小时通过酶联荧光测定法测量培养基中的孕酮水平。提取总RNA，用StAR蛋白、P450scc、HSD3B2、铁氧还蛋白和铁氧还蛋白还原酶的特异性引物进行实时定量RT-PCR。提取全细胞裂解物，用 StAR 蛋白抗体进行蛋白质印迹分析。经过 cAMP 处理后，培养基中的孕酮浓度增加至 38 倍。与cAMP治疗相比，CS显着降低了孕酮浓度30%（p<0.05），而胆固醇没有改变孕酮浓度。 CS处理使StAR mRNA和P450scc mRNA的表达分别下调54%和60%（p<0.05）。蛋白质印迹分析显示，CS 处理也减少了 StAR 蛋白的量。 cAMP处理后HSD3B2 mRNA的表达上调至3.4倍。铁氧还蛋白和铁氧还蛋白还原酶 mRNA 的表达不受 CS 处理的影响。这些数据表明CS通过调节StAR和P450scc基因的表达对孕酮产生具有抑制作用。
Cholesterol sulfate (CS) is a component of cell membranes that plays a role in stabilizing the cell membrane. We previously reported that CS increased in the endometrium during implantation, suggesting that CS plays an important role in reproduction. It has been reported that CS regulates progesterone and pregnenolone production in the placenta, adrenal glands and ovary. The regulatory mechanisms of steroid hormone production by CS, however, are still unknown. In the present study, we investigated the effect of CS on the expression of progesterone production-related genes in KGN cells, derived from human granulosa-like tumor. KGN cells were cultured with CS (10 muM) or cholesterol (10 muM) in the presence of 8-bromo-cAMP (1 mM). Progesterone levels in the culture media were measured by enzyme linked fluorescent assay at 24 h after treatment of CS and cAMP. Total RNAs were extracted for quantitative real time RT-PCR with specific primer of StAR protein, P450scc, HSD3B2, ferredoxin and ferredoxin reductase. Whole cell lysates were extracted for western blot analysis with antibody for StAR protein. Progesterone concentration in the culture medium increased to 38-fold by treatment of cAMP. CS significantly reduced progesterone concentration by 30% compared with those of cAMP treatment (p<0.05), while cholesterol did not change the progesterone concentration. CS treatment down-regulated the expression of StAR mRNA and P450scc mRNA was to 54% and 60%, respectively (p<0.05). Western blot analysis revealed that the amount of StAR protein was also reduced by CS treatment. The expression of HSD3B2 mRNA was up-regulated to 3.4-fold by treatment of cAMP. The expression of ferredoxin and ferredoxin reductase mRNA was not affected by CS treatment. These data implied that CS has an inhibitory effect on progesterone production by regulating the expression of StAR and P450scc gene expression.
硫酸胆固醇是离体大鼠肾上腺线粒体中天然存在的类固醇生成抑制剂。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Xu,XX;Lambeth,JD
通讯作者：
Lambeth,JD
乙醇对培养的大鼠颗粒细胞中卵泡刺激素诱导的类固醇生成急性调节蛋白（StAR）的影响。
DOI：
10.1016/j.alcohol.2006.01.001
发表时间：
2005
期刊：
Alcohol (Fayetteville, N.Y.)
影响因子：
--
作者：
Srivastava,VinodK;Vijayan,E;Hiney,JillK;Dees,WLes
通讯作者：
Dees,WLes
硫酸胆固醇在不同于细胞色素 P-450scc 的位点抑制肾上腺线粒体胆固醇侧链裂解。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lambeth,JD;Xu,XX;Glover,M
通讯作者：
Glover,M