骨桥蛋白在Ishikawa细胞的表达及调节

Expression and regulation of osteopontin in Ishikawa cell line

目的探讨骨桥蛋白（osteopontin，OPN）在Ishikawa细胞的表达及雌、孕激素、肝素结合表皮生长因子（HBEGF）对其表达的调节。方法体外培养高分化子宫内膜癌细胞Ishikawa细胞株，分别加入17-β雌二醇（E2）、孕激素（MPA）、MPA＋E2及HB-EGF处理48～72h后，采用原位杂交、逆转录聚合酶链反应（RT-PCR）、免疫印迹法（Western blot）测定各处理组OPN mRNA及其蛋白的表达。结果OPN mRNA（245bp）及其蛋白（45×10Z^3）在Ishikawa细胞均有表达；MPA、MPA＋E2和HB-EGF均可增加OPN mRNA的表达（P〈0．05），分别增加42％、32％及87％；OPN蛋白表达增加显著（P〈0．05），分别为127％、67％及163％，E2对其表达无明显影响。结论OPN在Ishikawa细胞的表达同时受孕激素和生长因子的上调。

Objective： To investigate the expression of osteopontin （OPN） and the regulatory effects of estradiol, progesterone and heparin-binding epidermal growth factor （HB-EGF） on OPN in Ishikawa cells. Methods： The high differentiated endometrial adenocarcinoma cells （Ishikawa cell line） were cultured in RPMI 1640 medium. After the cells were stimulated with estradiol （E2）, progesterone （MPA）, MPA in combination with E2 and HB-EGF, respectively, for 48-72 hours, the expression of OPN was detected by in situ hybridization, semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR） and Western blot analysis. Results： Both OPN mRNA and OPN protein （45KD） expressed in Ishikawa cell line. Compared to the control group, MPA, MPA in combination with E2 and HB-EGF increased the expression of OPN mRNA by 42%, 32%, 87% respectively （P〈0.05） and that of OPN protein by 127%, 67%, 163%, respectively （P〈0. 05）. Little effect was seen with the treatment of E2 alone （P〉0.05）. Conclusion： In endometrial epithelial cells,OPN is strongly induced by progesterone or HB-EGF.