人卵巢颗粒肿瘤细胞COV434中抗苗勒氏管激素抑制干细胞生长因子表达的实验研究

Inhibition of stem cell factor expression by anti-müllerian hormone in human ovarian granulosa cell tumor COV434 cell line: an experimental study

目的探讨人卵巢颗粒肿瘤细胞COV434中AMH对SCF表达的抑制作用。方法用课题组前期已构建及鉴定成功转染AMH真核表达载体p IRES2-EGFP-AMH的COV434细胞系作为实验组,将空质粒载体转染的COV434细胞系作为阴性对照组;未经转染的COV434系作为空白对照组,分别采用Western blot和免疫荧光法检测不同干预组SCF蛋白的表达水平。结果1Western blot实验结果显示转染AMH真核表达载体p IRES2-EGFP-AMH的COV434中AMH表达较空白对照组和阴性对照组明显增强(P<0.05),阴性对照组和空白对照组无统计学差异(P>0.05)。2Western blot实验结果显示SCF在实验组表达较空白对照组和阴性对照组显著降低(P<0.05)。3免疫荧光实验结果显示SCF在实验组的平均光密度值较阴性对照组及空白对照组的表达显著升高(P<0.05)。结论人卵巢颗粒肿瘤细胞COV434中AMH能有效抑制SCF蛋白的表达,为颗粒细胞中AMH调控SCF机制的研究提供科学基础。

Objective To explore the inhibitory effect of anti-mtlllerian hormone （AMH） on stem cell factor （SCF） expression in human ovarian granulosa cell tumor COV434 cell line. Methods Human ovarian granulosa cell tumor COV434 cell line was successfully constructed and identified in earlier stage, the cells transfected by AMH eukaryotic expression vector （plRES2-EGFP-AMH） were selected as experimental group, while the cells transfeeted by blank vector were selected as negative control group; the cells without transfection were selected as blank control group; Western blot and immunoflnorescence were used to detect the expression levels of SCF protein in different groups. Results Western blot showed that the expression of AMH in experimental group was statistically significantly higher than those in negative control group and blank control group （ P〈0. 05） , there was no statistically significant difference between negative control group and blank control group （ P〉0. 05 ） ; the expression of SCF in experimental group was statistically significantly lower than those in negative control group and blank control group （ P〈0.05 ） . Immunofluorescenee showed that the average optical density of SCF was statistically significantly higher than those in negative control group and blank control group （ P〈0.05 ） . Conclusion AMH can effectively inhibit SCF protein expression in human ovarian granulosa cell tumor COV434 cell line, which provides a scientific basis for studying the regulatory mechanism of AMH for SCF in human ovarian granulosa cell tumor COV434 cell line.