胰岛素和睾酮对Ishikawa细胞HOXAIO基因表达的影响

目的观察胰岛素（INS）和睾酮（T）对子宫内膜腺癌细胞HOXA10基因表达的影响。方法免疫细胞化学检测HOXA10蛋白在Ishikawa细胞定位表达；体外培养Ishikawa细胞，予不同浓度INS（90、60、30、3、0．3U／L）或T（10^-3、10^-4、10^-5、10^-5、10^-7mol／L）刺激Ishikawa细胞48h，MTT法检测INS、T对Ishikawa细胞生长的作用;最后分别以30U／LINS和10^-5mol／LT刺激Ishikawa细胞48h，逆转录聚合酶链反应（RT-PCR）方法测定INS和T对Ishikawa细胞HOXA10mRNA表达的影响。结果（1）HOXA10蛋白定位表达于Ishikawa细胞的细胞浆内。（2）不同浓度的INS均可促进Ishikawa细胞的生长，随着浓度的增加作用越强，INS浓度达60、90U／L时，细胞生长状况与浓度为30U／L相似。不同浓度的T均可抑制Ishikawa细胞的生长，随浓度增加抑制作用越明显，浓度达10^-4、10^-3mol／L时，细胞生长抑制状况与浓度为10^-3mol／L相似。（3）RT-PCR检测结果显示INS组和T组Ishikawa细胞中HOXA10mRNA表达均较对照组减弱（P〈0．01或P〈0．05）。结论不同浓度INS和T均可影响Ishikawa细胞生长，高浓度的INS和T降低细胞上HOXA10mRNA的表达。

The effects of insulin and testosterone on the expression of HOXAIO in Ishikawa cell

Objective：To investigate the effects of insulin and testosterone on the expression of HOXA10 in Ishikawa cell and explore the regulatory mechanism of HOXA10 in glandular epithelial cells of polycystic ovarian syndrome （PCOS）.
Methods： The expression of HOXA10 in Ishikawa cell was detected by immunocytochemistry. Ishikawa cells cultured in vitro were treated with different concentrations of exogenous insulin （90,60,30, 3,0.3 U/L） or testosterone （10^-3,10^-4,10^-5,10^-6,10^-7 mol/L）. The effects of insulin or testosterone on the growth of Ishikawa cells were detected by MTT. For the detection of the expression of HOXA10mRNA in Ishikawa cells, the cultured cells were treated with insulin （30 U/L） or testosterone （10^-5mol/L） for 48h and detected by RT-PCR, with the cells without any treatment as control.
Results：① The expression of HOXA10 in Ishikawa cells was localized in the intracytoplasm of Ishikawa cells by immunocytochemistry. （2）MTT method showed that insulin can stimulate the growth of Ishikawa cell in a dose-response manner at the concentrations of 0.3-30 U/L, with the maximal effect at concentration of 30 U/L, while testosterone can inhibit the cell growth in a dose-response manner and the effect of testosterone peaked at the concentration of 10^-5 mol/L. （3） RT-PCR demonstrated the aecreaseu HOXA10mRNA expression in Ishikawa cell after exposure to insulin 30 U/L or testosterone 10^-7 mol/L for 48 hours（P〈0.01, P〈0.05, respectively）.
Conclusions.Insulin or testosterone at different concentrations can affect the growth of Ishikawa cell. The expression of HOXA10mRNA in Ishikawa cells decreased after the treatment of high concentration of insulin or testosterone.