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目的:探讨特异AT序列结合蛋白1(specialAT-rich sequence-bindingprotein,SATB1)在卵泡刺激素(Follicle stimulating hormone,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-time PCR检测不同浓度FSH(0、10、20、40、80mIU/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-Negative contro1)序列的实验组,对SATB1无干扰作用;②siSATB1组:转染特异性干扰下调SATB1的siSATB1序列;③FSH+siCon组:以FSH处理的siCon组;④FSH+siSATB1组:以FSH处理的siSATB1组。MTT法检测4组细胞的增殖情况,Western blotting技术检测4组细胞细胞周期蛋白(CyclinD1),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Cyclin D1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(302 12)个明显高于siCon组(139 19)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,SATB1mRNA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/ml时最显著(P<0.05)。4.FSH+siSATB1组的细胞增殖能力明显低于FSH+siCon组的细胞增殖能力,FSH+siSATB1组的Cyclin D1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(52 16)个低于FSH+siCon组的(302 12)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:SATB1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。
Objective: To investigate the role of special AT-1 (SATB1) in the proliferation and invasion of epithelial ovarian cancer ES-2 cells induced by Follicle stimulating hormone (FSH). Methods: Real-time PCR was used to detect the mRNA expression level of SATB1 after treatment with different concentrations of FSH (0, 10, 20, 40 and 80 mIU / ml). The experiment was divided into four groups: ①siCon group, the experimental group transfected with si-negative contro1 sequence had no interference with SATB1; ②siSATB1 group: transfected with specific siRNA to down-regulate the siSATB1 sequence of SATB1; ③FSH + siCon group: FSH-treated siCon group; FSH + siSATB1 group: siSATB1 treated with FSH. The proliferation of 4 groups of cells was detected by MTT assay. The protein expression of CyclinD1 and MMP-2 were detected by Western blotting. The invasion ability of 4 groups was detected by Transwell invasion assay. The proliferation of FSH + siCon group was significantly higher than that of siCon group. The relative expression of Cyclin D1 protein in FSH + siCon group was significantly higher than that in siCon group (0.90 ± 0.08 vs 0.37 ± 0.01, all P <0.01 ), Suggesting that FSH can promote the proliferation of ES-2 cells. The number of transmembrane cells in FSH + siCon group (302 12) was significantly higher than that in siCon group (139 19). The relative expression level of MMP-2 protein in FSH + siCon group was significantly higher than that in siCon group (0.28 ± 0.40 ± 0.01) 0.02, suggesting that FSH has the ability to promote ES-2 cell invasion. The expression of SATB1mRNA increased gradually with the increase of FSH concentration, which was respectively1.1.66 ± 0.04,1.79 ± 0.21,2.31 ± 0.03. When FSH concentration was80mlU / ml, the expression of SATB1mRNA was the most significant (P <0.05). The cell proliferation ability of FSH + siSATB1 group was significantly lower than that of FSH + siCon group, the relative expression level of Cyclin D1 protein in FSH + siSATB1 group was significantly lower than that in FSH + siCon group (0.22 ± 0.02 vs 0.90 ± 0.08, P < <0.01). The number of transmembrane cells (52 16) in FSH + siSATB1 group was lower than that in FSH + siCon group (302 12). The relative expression of MMP-2 protein in FSH + siSATB1 group was significantly lower than that of FSH + siCon 0.40 ± 0.01 (P <0.01). The ability of FSH to promote the proliferation and invasion of ES-2 cells was blocked by the decrease of SATB1 gene expression. Conclusion: SATB1 is an important target of FSH and mediates the regulation of FSH on the proliferation and invasion of epithelial ovarian cancer cell line ES-2.