Pan Tianying, He Guolin, Chen Meng, Bao Chenyi, Chen Yan, Liu Guangyu, Zhou Mi, Li Shuying, Xu Wenming, Liu Xinghui
Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu 610041, China.
Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu 610041, China.
Oncotarget. 2017 Sep 22;8(52):89824-89836. doi: 10.18632/oncotarget.21158. eCollection 2017 Oct 27.
In this study, we investigated the exact mechanism by which excessive CYP11A1 expression impairs the placentation process and whether this causes preeclampsia (PE) in an model.
In order to study CYP11A1 overexpression, BeWo cells were transfected with CYP11A1. Pregnenolone, progesterone, and testosterone levels were measured by enzyme linked immunosorbent assays, and levels of autophagy markers were quantified by western blotting and immunofluorescence. Trophoblastic cell invasion was assessed using transwell assays; BeWo cells were treated with testosterone and an androgen receptor (AR) inhibitor (flutamide) to elucidate the invasion mechanism. An adenovirus overexpression rat model was established to investigate CYP11A1 overexpression and the phenotype was examined. Furthermore, human placenta samples ( = 24) were used to determine whether PE patient placentas showed altered CYP11A1 and autophagy marker expression.
BeWo cells overexpressing CYP11A1 had significantly increased levels of pregnenolone, progesterone, and testosterone. Additionally, the expression levels of autophagy markers in CYP11A1-overexpressing BeWo cells were significantly increased. Trophoblast invasion was significantly reduced in CYP11A1-overexpressing cells as well as in cells treated with high testosterone. This reduction could be significantly rescued when cells were pretreated with flutamide. Overexpression of CYP11A1 in rat pregnancies led to PE-like symptoms and an over-activation of the AR-mediated pathway in the placenta. Elevated expression of CYP11A1 and autophagy markers could also be detected in PE placenta samples.
These results suggest that abnormally high expression of CYP11A1 induces trophoblast autophagy and inhibits trophoblastic invasion, which is associated with the etiology of PE.
在本研究中，我们调查了CYP11A1过度表达损害胎盘形成过程的确切机制，以及这是否会在大鼠模型中导致子痫前期（PE）。
为了研究CYP11A1的过表达，用CYP11A1转染BeWo细胞。通过酶联免疫吸附测定法测量孕烯醇酮、孕酮和睾酮水平，并通过蛋白质印迹法和免疫荧光法定量自噬标志物的水平。使用Transwell测定法评估滋养层细胞侵袭；用睾酮和雄激素受体（AR）抑制剂（氟他胺）处理BeWo细胞以阐明侵袭机制。建立腺病毒过表达大鼠模型以研究CYP11A1的过表达，并检查其表型。此外，使用人胎盘样本（n = 24）来确定PE患者胎盘是否显示CYP11A1和自噬标志物表达的改变。
过表达CYP11A1的BeWo细胞中孕烯醇酮、孕酮和睾酮水平显著升高。此外，过表达CYP11A1的BeWo细胞中自噬标志物的表达水平显著增加。过表达CYP11A1的细胞以及用高剂量睾酮处理的细胞中滋养层侵袭显著降低。当细胞用氟他胺预处理时，这种降低可以得到显著挽救。大鼠妊娠中CYP11A1的过表达导致类似PE的症状以及胎盘中AR介导的途径过度激活。在PE胎盘样本中也可以检测到CYP11A1和自噬标志物的表达升高。
这些结果表明，CYP11A1异常高表达诱导滋养层自噬并抑制滋养层侵袭，这与PE的病因有关。
