Wang Jiayin, Ohara Noriyuki, Wang Zhuo, Chen Wei, Morikawa Akira, Sasaki Hiroko, DeManno Deborah A, Chwalisz Kristof, Maruo Takeshi
Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine, Kobe, Japan.
Hum Reprod. 2006 Jul;21(7):1869-77. doi: 10.1093/humrep/del035. Epub 2006 Apr 13.
This study was conducted to evaluate the effects of a novel selective progesterone receptor modulator (SPRM) asoprisnil on the expression of growth factors and their receptors and on growth factor-induced proliferation of cultured uterine leiomyoma and matching myometrial cells.
The expression of epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I) and transforming growth factor (TGFbeta3) was assessed by immunocytochemistry and semi-quantitative RT-PCR. The expression of phosphorylated EGF receptor (p-EGFR), IGF-I receptor alpha subunit (IGF-IRalpha) and phosphorylated TGFbeta receptor type II (p-TGFbeta RII) was assessed by Western blot analysis. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay.
Treatment with 10(-7) M asoprisnil decreased EGF, IGF-I and TGFbeta3 mRNA and protein expression as well as p-EGFR, IGF-IRalpha and p-TGFbeta RII protein expression in leiomyoma cells cultured for 72 h. EGF (100 ng/ml), IGF-I (100 ng/ml) and TGFbeta3 (10 ng/ml) increased the number of viable leiomyoma cells cultured for 72 h, whereas the concomitant treatment with 10(-7) M asoprisnil antagonized the growth factor-induced increase in leiomyoma cell proliferation. In cultured myometrial cells, however, asoprisnil affected neither the growth factor and their receptor expression nor the cell proliferation.
Asoprisnil inhibits the expression of EGF, IGF-I, TGFbeta3 and their receptors in cultured leiomyoma cells without affecting their expressions in myometrial cells.
本研究旨在评估新型选择性孕激素受体调节剂(SPRM)阿索普瑞尼对生长因子及其受体表达以及生长因子诱导的培养子宫平滑肌瘤和配对子宫肌层细胞增殖的影响。
通过免疫细胞化学和半定量逆转录聚合酶链反应(RT-PCR)评估表皮生长因子(EGF)、胰岛素样生长因子-I(IGF-I)和转化生长因子(TGFβ3)的表达。通过蛋白质印迹分析评估磷酸化表皮生长因子受体(p-EGFR)、IGF-I受体α亚基(IGF-IRα)和磷酸化转化生长因子β受体II型(p-TGFβRII)的表达。通过3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑𬭩测定法评估细胞增殖。
用10⁻⁷M阿索普瑞尼处理72小时培养的平滑肌瘤细胞后,EGF、IGF-I和TGFβ3的mRNA和蛋白表达以及p-EGFR、IGF-IRα和p-TGFβRII蛋白表达降低。EGF(100 ng/ml)、IGF-I(100 ng/ml)和TGFβ3(10 ng/ml)增加了72小时培养的存活平滑肌瘤细胞数量,而同时用10⁻⁷M阿索普瑞尼处理可拮抗生长因子诱导的平滑肌瘤细胞增殖增加。然而,在培养的子宫肌层细胞中,阿索普瑞尼既不影响生长因子及其受体表达,也不影响细胞增殖。
阿索普瑞尼抑制培养的平滑肌瘤细胞中EGF、IGF-I、TGFβ3及其受体的表达,而不影响子宫肌层细胞中的表达。