Shi Xiu, Xu Wei, Sun Ying, Dai Huihua, Wang Xiuli
Department of Gynecology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210036, China.
Department of Gynecology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210036, China. Email:
Zhonghua Fu Chan Ke Za Zhi. 2014 Feb;49(2):114-9.
To study the expression patterns of steroid receptor coactivators (SRC) and steroid-induced stromal cell-derived factor-1 (SDF-1) in endometriosis, and to explore the roles of SRC in the steroid-induced SDF-1 expression endometriosis.
From May 2010 to October 2012, 16 endometriosis cases at stages III or IV according to the revised American Society for Reproductive Medicine classification undergoing surgery in the First Affiliated Hospital to Nanjing Medical University were enrolled in this study. Their ectopic endometrium were from ovarian endometriomata which were identified pathologically with 9 cases at proliferative phase and 7 cases at secretory phase. The normal endometrium were acquired from the healthy women with normal menstrual cycle (n = 10, proliferative phase = 5, secretory phase = 5). The mRNA levels of SRC and SDF-1α during the menstrual cycle were detected by quantitative real-time polymerase chain reaction. Ectopic endometrium stromal cells were purified and cultured in medium containing 17β-estradiol (10(-8) mol/L) or 17β-estradiol (10(-8) mol/L) + progesterone (10(-6) mol/L). At 24, 48, 72 and 96 hours, the supernatants were collected to measure SDF-1α expression by ELISA. Ectopic endometrium stromal cells were transfected respectively with siRNA of SRC-1 and SRC-2 using lipofectamine. Two days after transfection, 17β-estradiol (10(-8) mol/L) or 17β-estradiol (10(-8) mol/L) + progesterone (10(-6) mol/L) were added into the media. On the third day after the steroid hormones treatment, the media were collected to quantify SDF-1α expression with ELISA.
(1) Cyclical changes: the SRC-1, SRC-2 and SDF-1α showed marked cyclic differences in normal endometrium (P < 0.05). In proliferative phase and secretory phase, the SRC-1, SRC-2 and SDF-1α were 5.6 ± 1.2, 3.8 ± 1.1, 2.7 ± 0.5 and 2.6 ± 1.0, 2.1 ± 1.0, 1.6 ± 0.5, respectively. There was no periodic variation in the expression of SRC-1, SRC-2 and SDF-1α in ectopic endometrium throughout the menstrual cycle. (2) Steroid-induced SDF-1α expression in ectopic endometrium stromal cells: the 17β-estradiol-induced SDF-1α expression was (1 803 ± 196), (2 272 ± 261) and (2 162 ± 258) ng/L at 48, 72 and 96 hours . At the same time points, the SDF-1α expression induced by 17β-estradiol and progesterone was (1 307 ± 150), (1 518 ± 301) and (1 550 ± 144) ng/L, respectively. There was significant difference between two groups (P < 0.05 ). (3) The effects of SRC silencing on steroid hormones-induced SDF-1α expression in ectopic endometrium stromal cells: the expression of 17β-estradiol-induced SDF-1α at 72 hours was significantly decreased from (2 313 ± 357) ng/L to (1 155 ± 244) ng/L after the silencing of SRC-1 (P < 0.05). After the silencing of SRC-2, the 17β-estradiol-induced SDF-1α at 72 hours was (1 958 ± 324) ng/L. There was no significant difference compared with the before the silencing (P > 0.05). The expression of SDF-1α at 72 hours induced by 17β-estradiol+progesterone was (1 534 ± 449) ng/L and (2 051 ± 380) ng/L respectively before and after the silencing of SRC-2 and showed the significant difference (P < 0.05).
During the expression of SDF-1α regulated by steroids in ectopic endometrium cells, SRC-1 is the major coactivator of 17β-estradiol and SRC-2 is the major coactivator of progesterone.
研究类固醇受体共激活因子（SRC）和类固醇诱导的基质细胞衍生因子-1（SDF-1）在子宫内膜异位症中的表达模式，并探讨SRC在类固醇诱导的子宫内膜异位症中SDF-1表达中的作用。
2010年5月至2012年10月，选取南京医科大学第一附属医院收治的16例按照美国生殖医学学会修订分类法为III或IV期的子宫内膜异位症患者行手术治疗。其异位内膜取自卵巢子宫内膜异位囊肿，经病理确诊，其中增殖期9例，分泌期7例。正常子宫内膜取自月经周期正常的健康女性（n = 10，增殖期 = 5，分泌期 = 5）。采用定量实时聚合酶链反应检测月经周期中SRC和SDF-1α的mRNA水平。纯化异位内膜基质细胞，在含有17β-雌二醇（10⁻⁸ mol/L）或17β-雌二醇（10⁻⁸ mol/L）+孕酮（10⁻⁶ mol/L）的培养基中培养。在24、48、72和96小时时，收集上清液，采用酶联免疫吸附测定法检测SDF-1α表达。采用脂质体分别将SRC-1和SRC-2的小干扰RNA转染至异位内膜基质细胞。转染后两天，向培养基中加入17β-雌二醇（10⁻⁸ mol/L）或17β-雌二醇（10⁻⁸ mol/L）+孕酮（10⁻⁶ mol/L）。在类固醇激素处理后的第三天，收集培养基，采用酶联免疫吸附测定法对SDF-1α表达进行定量。
（1）周期性变化：正常子宫内膜中SRC-1、SRC-2和SDF-1α呈现明显的周期性差异（P < 0.05）。在增殖期和分泌期，SRC-1、SRC-2和SDF-1α分别为5.6 ± 1.2、3.8 ± 1.1、2.7 ± 0.5和2.6 ± 1.0、2.1 ± 1.0、1.6 ± 0.5。异位内膜中SRC-1、SRC-2和SDF-1α的表达在整个月经周期中无周期性变化。（2）类固醇诱导的异位内膜基质细胞中SDF-1α表达：17β-雌二醇诱导的SDF-1α在48、72和96小时时的表达分别为（1 803 ± 196）、（2 272 ± 261）和（2 162 ± 258）ng/L。在相同时间点，17β-雌二醇和孕酮诱导的SDF-1α表达分别为（1 307 ± 150）、（1 518 ± 301）和（1 550 ± 144）ng/L。两组间差异有统计学意义（P < 0.05）。（3）SRC沉默对类固醇激素诱导的异位内膜基质细胞中SDF-1α表达的影响：SRC-1沉默后，17β-雌二醇诱导的72小时SDF-1α表达从（2 313 ± 357）ng/L显著降低至（1 155 ± 244）ng/L（P < 0.05）。SRC-2沉默后，17β-雌二醇诱导的72小时SDF-1α表达为（1 958 ± 324）ng/L。与沉默前相比差异无统计学意义（P > 0.05）。SRC-2沉默前后，17β-雌二醇+孕酮诱导的72小时SDF-1α表达分别为（1 534 ± 449）ng/L和（2 051 ± 380）ng/L，差异有统计学意义（P < 0.05）。
在异位内膜细胞中类固醇调节的SDF-1α表达过程中，SRC-1是17β-雌二醇的主要共激活因子，SRC-2是孕酮的主要共激活因子。
