钠依赖性维生素C转运子2的维生素C摄取可通过绒毛膜癌细胞中的Sp1和TFAP2A转录因子诱导βhCG表达

Wu, X.; Department of Pharmacology, School of Medicine, Zhejiang University, No. 866, Yuhangtang Road, Hangzhou 310058, China; email:xiwu@zju.edu.cn

Context: Vitamin C [ascorbic acid (AA)] is transported by sodium-dependent vitamin C transporters (SVCT) 1 and 2, and our previous studies show AA induces a dramatic production of steroid hormones in human choriocarcinoma cells. However, whether AA induces the production of placental polypeptide hormones remains unknown. Here we investigated the mechanisms governing AA-induced β-human chorionic gonadotropin (hCG) expression. Methods: Frozen sections from human term placentas were used for immunostaining of SVCT, and βhCG mRNA expression and its production in primary human placental cytotrophoblasts and JEG-3 cells were examined by quantitative RT-PCR and ELISA, respectively. Knockdown of SVCT2, transcription factor activating enhancer-binding protein 2α (TFAP2A), or specificity protein-1 (Sp1) expression was achieved by retrovirus-mediated short hairpin RNA, and the transcriptional factors responsible for AA-induced βhCG expression was identified by reporter constructs. Results: Both SVCT1 and SVCT2 are expressed in human term placentas. SVCT2 is predominantly localized in the syncytial layer, whereas SVCT1 is predominantly distributed in the villous core. AA dramatically induces βhCG mRNA expression and its production in JEG-3 cells and primary human cytotrophoblasts, and knockdown of SVCT2 expression in JEG-3 cells significantly decreases AA-induced βhCG expression. Data from βhCG5 construct and its deletion mutants further indicate that AA induces βhCG5 transactivation through Sp1 and TFAP2A transcriptional factors, and silence of Sp1 and/or TFAP2A expression significantly decreased AA-induced βhCG5 reporter activity and βhCG expression as well. Conclusions: The present study revealed the novel effects of AA on polypeptide hormone, βhCG, production and the potential mechanisms governing AA-induced βhCG expression, suggesting the potentially indispensable roles of AA in placental endocrine and pregnant maintenance. Copyright © 2012 by The Endocrine Society.

Department of Pharmacology, School of Medicine, Zhejiang University, No. 866, Yuhangtang Road, Hangzhou 310058, China