Activation of SRC kinase and phosphorylation of signal transducer and activator of transcription-5 are required for decidual transformation of human endometrial stromal cells
Activation of SRC kinase and phosphorylation of signal transducer and activator of transcription-5 are required for decidual transformation of human endometrial stromal cells
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DOI：
10.1210/en.2007-1217
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发表时间：
2008-03-01
期刊：
影响因子：
4.8
通讯作者：
Yoshimura, Yasunori
中科院分区：
文献类型：
--
作者：
Nagashima, Takashi;Maruyama, Tetsuo;Yoshimura, Yasunori
孕酮通过激活蛋白激酶A、信号转导和转录激活因子5等多种信号通路，诱导雌激素诱导的人子宫内膜基质细胞蜕膜转化，而蜕膜基质细胞对着床和妊娠的维持至关重要。我们先前已经证明，v-src肉瘤(Schmidt-Ruppin A-2)病毒癌基因同源(SRC)激酶的激活与蜕膜化密切相关，SRC对于小鼠最大程度的蜕膜化是不可或缺的。为了研究SRC激酶活性是否对人类蜕膜化是必不可少的，我们用携带增强型绿色荧光蛋白的腺病毒(Ad-EGFP)、不活跃的显性-阴性突变株(Ad-SRC/K295R)或不活跃的自身磷酸化位点突变株(Ad-SRC/Y416F)感染人胚胎干细胞。在雌二醇和孕酮(EP)的存在下培养细胞以诱导蜕膜形成，并进行RT-PCR、免疫印迹和ELISA法分析。Ad-EGFP感染的人胚胎干细胞发生蜕膜转化，蜕膜化标志物IGF结合蛋白1和催乳素在EP处理12天后表达上调。相反，感染Ad-SRC/K295R的人胚胎干细胞即使在EP处理后仍保持成纤维样细胞形态，不产生IGF结合蛋白1和催乳素。Ad-SRC/Y416F对蜕膜化的抑制作用与Ad-SRC/K295R相似，但抑制作用较弱。在蜕膜化过程中，STAT5在酪氨酸694上被磷酸化，酪氨酸694是众所周知的SRC磷酸化位点。Ad-SRC/K295R组的磷酸化水平显著降低，而Ad-EGFP组的磷酸化水平无明显变化。这些结果表明，SRC-STAT5通路是hESCs蜕膜形成所必需的。
Progesterone induces decidual transformation of estrogen-primed human endometrial stromal cells (hESCs), critical for implantation and maintenance of pregnancy, through activation of many signaling pathways involving protein kinase A and signal transducer and activator of transcription (STAT)-5. We have previously shown that kinase activation of v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (SRC) kinase is closely associated with decidualization and that SRC is indispensable for maximal decidualization in mice. To address whether SRC kinase activity is essential for decidualization in humans, hESCs were infected with adenoviruses carrying enhanced green fluorescent protein alone (Ad-EGFP), a kinase-inactive dominant-negative mutant (Ad-SRC/K295R), or an inactive autophosphorylation site mutant (Ad-SRC/Y416F). The cells were cultured in the presence of estradiol and progesterone (EP) to induce decidualization and subjected to RT-PCR, immunoblot, and ELISA analyses. Ad-EGFP-infected hESCs exhibited decidual transformation and up-regulation of decidualization markers including IGF binding protein 1 and prolactin in response to 12-d treatment with EP. In contrast, hESCs infected with Ad-SRC/K295R remained morphologically fibroblastoid without production of IGF binding protein 1 and prolactin even after EP treatment. Ad-SRC/Y416F displayed similar but less inhibitory effects on decidualization, compared with Ad-SRC/K295R. During decidualization, STAT5 was phosphorylated on tyrosine 694, a well-known SRC phosphorylation site. Phosphorylation was markedly attenuated by Ad-SRC/K295R but not Ad-EGFP. These results indicate that the SRC-STAT5 pathway is essential for decidualization of hESCs.