Peluso John J, Gawkowska Anna, Liu Xiufang, Shioda Toshi, Pru James K
Department of Cell Biology, University of Connecticut Health Center, Farmington, Connecticut 06030, USA.
Endocrinology. 2009 Nov;150(11):4846-54. doi: 10.1210/en.2009-0730. Epub 2009 Oct 1.
To determine whether progesterone receptor membrane component 1 (PGRMC1) regulates the development and cisplatin (CDDP)-sensitivity of human ovarian tumors, PGRMC1 was depleted from a human ovarian cancer cell line, dsRed-SKOV-3 cells, using a short hairpin RNA knockdown approach. Compared with parental dsRed-SKOV-3 cells, the PGRMC1-deplete cells grew slower in vitro and did not show progesterone's (P4) antiapoptotic effect. In fact, P4 induced apoptosis in PGRMC1-deplete cells in a dose-dependent manner. When transplanted into the peritoneum of athymic nude mice, parental dsRed-SKOV-3 cells developed numerous tumors, which were classified as either typical or oxyphilic clear cell tumors. CDDP increased the percentage of apoptotic nuclei in typical clear cell tumors and P4 attenuated CDDP-induced apoptosis. In contrast, the percentage of apoptotic nuclei in oxyphilic clear cell tumors was low (< or =1%) and was not significantly affected by CDDP and/or P4. Compared with tumors derived from parental dsRed SKOV-3 cells, PGRMC1-deplete tumors: 1) developed in fewer mice, 2) formed less frequently, 3) appeared smaller, and 4) resulted in fewer oxyphilic clear cell tumors. These PGRMC1-deplete tumors were not responsive to CDDP's apoptotic effects. The failure to respond to CDDP could be due to their poorly developed microvasculature system as judged by percentage of CD31-stained endothelial cells and/or their increased expression of ATP-binding cassette transporters, which are involved in drug resistance. Taken together, these findings indicate that PGRMC1 plays an essential role in the development and CDDP sensitivity of human ovarian tumors.
为了确定孕激素受体膜成分1（PGRMC1）是否调节人类卵巢肿瘤的发生发展和顺铂（CDDP）敏感性，采用短发夹RNA敲低方法，从人卵巢癌细胞系dsRed-SKOV-3细胞中去除PGRMC1。与亲代dsRed-SKOV-3细胞相比，PGRMC1缺失的细胞在体外生长较慢，且未表现出孕激素（P4）的抗凋亡作用。事实上，P4以剂量依赖的方式诱导PGRMC1缺失细胞凋亡。当移植到无胸腺裸鼠的腹膜中时，亲代dsRed-SKOV-3细胞形成了许多肿瘤，这些肿瘤被分类为典型或嗜酸性透明细胞肿瘤。CDDP增加了典型透明细胞肿瘤中凋亡细胞核的百分比，而P4减弱了CDDP诱导的凋亡。相比之下，嗜酸性透明细胞肿瘤中凋亡细胞核的百分比很低（≤1%），且不受CDDP和/或P4的显著影响。与源自亲代dsRed SKOV-3细胞的肿瘤相比，PGRMC1缺失的肿瘤：1）在较少的小鼠中发生，2）形成频率较低，3）看起来较小，4）嗜酸性透明细胞肿瘤较少。这些PGRMC1缺失的肿瘤对CDDP的凋亡作用无反应。对CDDP无反应可能是由于其发育不良的微血管系统（根据CD31染色内皮细胞的百分比判断）和/或其参与耐药的ATP结合盒转运蛋白表达增加。综上所述，这些发现表明PGRMC1在人类卵巢肿瘤的发生发展和CDDP敏感性中起重要作用。
