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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、0.3 U/L)或T(10~(-3)、10~(-4)、10~(-5)、10~(-6)、10~(-7)mol/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30 U/LINS和10~(-5)mol/L T刺激Ishikawa细胞48 h,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、90 U/L时,细胞生长状况与浓度为30 U/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~(-3)mol/L时,细胞生长抑制状况与浓度为10~(-5)mol/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。
Objective To investigate the effect of insulin and testosterone (T) on HOXA10 gene expression in endometrial adenocarcinoma cells. Methods Immunocytochemistry was used to detect the expression of HOXA10 protein in Ishikawa cells. Ishikawa cells were cultured in vitro and treated with various concentrations of INS (90, 60, 30, 3, 0.3 U / L) or T (10 -3, Ishikawa cells were stimulated with 4, 10, 5, 10 -6, 10 -7 mol / L for 48 h. The effects of INS and T on the growth of Ishikawa cells were detected by MTT assay. Finally, Ishikawa cells were stimulated with 10 -5 mol / L of L-lysine for 48 h, and the effect of INS and T on HOXA10 mRNA expression in Ishikawa cells was determined by reverse transcription-polymerase chain reaction (RT-PCR). Results (1) HOXA10 protein was localized in the cytoplasm of Ishikawa cells. (2) Different concentrations of INS could promote the growth of Ishikawa cells. The stronger the effect was, the higher the INS concentration was 60,90 U / L, the similar to the concentration of 30 U / L. Different concentrations of T can inhibit the growth of Ishikawa cells, with the concentration increasing the more obvious inhibition, the concentration of 10 -4 and 10 -3 mol / L, the cell growth inhibition and concentration of 10 ~ ( -5) mol / L. (3) The results of RT-PCR showed that HOXA10mRNA expression in Ishikawa cells of INS group and T group was weaker than that of control group (P <0.01 or P <0.05). Conclusion Both INS and T at different concentrations can affect the growth of Ishikawa cells. High concentrations of INS and T decrease the expression of HOXA10 mRNA.