FSH通过SATB1调控上皮性卵巢癌ES-2细胞的增殖和侵袭活性

目的：探讨特异AT序列结合蛋白1（special AT-rich sequence-bindingprotein，SATB1）在卵泡刺激素（Follicle stimulating hor-mone，FSH）诱导的上皮性卵巢癌ES-2细胞增殖和侵袭中的作用。方法：以Real-timePCR检测不同浓度FSH（0、10、20、40、80mlU／ml）处理后sATB1基因mRNA表达水平的变化。实验分4组：①siCon组，转染si-阴性对照（si-Negativecontrol）序列的实验组，对SATBl无干扰作用；②siSATB1组：转染特异性干扰下调SATB1的siSATB1序列；（3）FSH＋siCon组：以FSH处理的siCon组；（4）FSH＋siSATBl组：以FSH处理的sisATB1组。MTT法检测4组细胞的增殖情况，Westernblotting技术检测4组细胞细胞周期蛋白（CyclinDl），基质金属蛋白酶2（MMP．2）的蛋白表达情况，Transwell侵袭实验检测4组细胞侵袭能力的变化。结果：1．FSH＋siCon组的细胞增殖能力明显高于siCon组的细胞增殖能力，FSH＋siCon组的CyclinD1蛋白相对表达量0．90＋0．08明显高于siCon组的0．37＋0．01（P均〈0．01），提示FSH具有促进ES．2细胞增殖的作用。2．FSH＋siCon组的穿膜细胞数（30212）个明显高于siCon组（13919）个，FSH＋siCon组的MMP．2蛋白相对表达量0．40＋0．01明显高于siCon组的0．28＋0．02，提示FSH具有促进ES．2细胞侵袭能力的作用。3．随着FsH浓度的增高，SATBlmRNA的表达量逐渐增加，分别为1，1．66±0．04，1．79±0．21，2.31±0．03，以FsH浓度为80mlU／m1时最显著（P〈0．05）。4．FSH＋siSATBl组的细胞增殖能力明显低于FSH＋siCon组的细胞增殖能力，FSH＋siSATBl组的CyclinD1蛋白相对表达量0．22±0．02明显低于FSH＋siCon组的0．90±0．08（P均〈0．01）；FSH＋siSATB1组的穿膜细胞数（5216）个低于FSH＋siCon组的（30212）个，FSH＋siSATB1组的MMP-2蛋白相对表达量0．15±0．00明显低于FSH＋siCon组的0．40±0．01（P均〈0．01），FSH促进ES-2细胞增殖和侵袭的能力由于SATB1基因表达的下降而被阻断。结论：SATBl是FSH作用的重要靶分子，介导FSH对上皮性卵巢癌ES-2细胞系增殖、侵袭活性的调控。

Follicle Stimulatinghormone Controls Proliferation and Invasion Activity of Epithelial Ovarian Cancer ES-2 Cells by Specialat-Rich Sequence-Binding Protein 1

Objective： To investigate the relationship between SATB1 （special AT-rich sequence-binding protein）and FSH （Follicle stimulating hormone ）,and their roles in regulating cell proliferation,invasion in epithelial ovarian carcinoma cells ES-2. Methods： Real-time PCR detects the expression levels of SATB1 in ES-2 cells after treatment with different concentrations of FSH （0,10,20,40,80mlU/ml）.ES-2 cells were divided into 4 groups：siCon group：transfected si-Negative control sequence which had no effect on SATB1 ;siSATB1 group：transfected siSATBl-specific sequence which down-regulated SATB1 ;FSH＋siCon group：siCon cells treated by FSH;FSH＋siSATB 1 ：siSATB 1 cells treated by FSH. MTT assay was used to determine cellular proliferative activity. Western blotting were used to determine the effect of FSH and SATB1 on the expression of cyclinD1 and MMP-2 in protein levels.The invasive activity was observed through matrigel invasion assay. Results： 1. The cell proliferation of FSH＋siCon was higher than that of siCon.The protein expression of Cyclin D 1 in FSH＋siCon was higher than that in siCon. （ P〈0.01）.It showed that FSH induced cell proliferation of ES-2.2. The numbers of ES-2 that penetrated the basement menbrane in FSH＋siCon were more than that in siCon. The protein expression of MMP-2 in FSH＋siCon was higher than that in siCon. （ P〈0.01）.It suggested that FSH induced cell invasion of ES-2.3.The expression of SATB1 increased as increasing in the concentration of FSH,the results were： 1,1.66±0.04, 1.79± 0.21,2.31 ± 0.03 （P〈0.05）. FSH up-regulated the expression of SATB1 in a dose-dependent manner.4. The cell proliferation in FSH＋siSATB1 was lower than that in FSH＋siCon.The protein expression of Cyclin D1 in FSH＋siSATB1 was lower than that in FSH＋siCon. （ P〈0.01）,the same as in cell invasion. Inhibition of SATB1 by RNA interference blocked the proliferation and invasion of ES-2 induced by FSH.Conelusions： FSH regulates cell proliferation and invasion by SATB 1 gene.