Earticle

초록

영어

The monkey 20ɑ-hydroxysteroid dehydrogenase (20ɑ-HSD) is an enzyme responsible for the catabolism in converting progesterone into biological inactive 20a-hydroxyprogesterone, thus playing a key role during the estrous cycle or pregnancy and allowing ovulation and parturition to occur in most mammalian animals. We have previously shown that 20ɑ-HSD in pre-ovulation and pre-parturition was highly detected in ovarian and placental tissues and localized mainly in the syncytiotrophoblast of the placenta. In this study, we focus on the analysis of molecular characterization of the monkey 20ɑ-HSD promoter region by using reporter assay systems in a Chinese hamster ovary (CHO) K1 cells. A reporter assay, using constructs (—890-Luc, —513-Luc, —306-Luc, —273-Luc and —70-Luc) of various lengths of the 5'-flanking region, revealed that the region (Ap-1) between —281 and —274 bp was essential for transcriptional activity. The absence of Ap-1 site in -273-Luc dramatically decreased to the control levels. Thus, Ap-1 site (—281→—274) (5'-TGTCTCAT-3') plays a crucial role for monkey 20α-HSD gene transcription. Next, we analyzed the monkey 20α-HSD promoter activity with treatment of prolactin, PGF2α and Forskolin in CHO-K1 cells. The 2005bp monkey promoter-pGL3 was transfected to CHO cells and added various dose-dependent treatments after incubation 24 hours. For PGF2α 0.5, 1, 5, 10 and 50 μM added and incubated during 6 hours. For Forskolin 0.1, 1, 10, 50 and 100 μM added. And finally, the luciferase activity was measured after 24 hours. Also, human prolactin (3μg/ml) was used as inhibitor. In result, PGF2a was response to a dose-dependent activation. On the other hand Forskolin was detected dose-dependent inhibition in the 2005bp 20α-HSD promoterpGL3 construct. However, prolactin was not shown any significant different. Taken together, our results indicate that monkey 20α-HSD promoter activity inhibit by forskolin. Also, PGF2a remarkedly stimulates the activity of monkey 20α-HSD promoter. We demonstrated that monkey 20a-HSD promoter regulate by AP-transcription factor, PGF2a, and Foskolin.

키워드

저자

• Eun-Bi Seo [ Department of Animal Biotechnology, Graduate School of Future Fusion Technology, Hankyong National University ]
• Tsevelmaa Nanjidsuren [ Department of Animal Biotechnology, Graduate School of Future Fusion Technology, Hankyong National University ]
• Chae-Won Park [ Department of Animal Biotechnology, Graduate School of Future Fusion Technology, Hankyong National University ]
• Min-Su Kim [ Department of Animal Biotechnology, Graduate School of Future Fusion Technology, Hankyong National University ]
• Kwan-Sik Min [ Department of Animal Biotechnology, Graduate School of Future Fusion Technology, Hankyong National University ]

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간행물

표지보기 관심저널 등록

• 간행물명

  Reproductive & Developmental Biology(Supplement)

• 간기

  연간

• 수록기간

  2001~2017

• 십진분류

  KDC 527 DDC 636