Full Text:   <1385>

Summary:  <996>

CLC number: S828.2

On-line Access: 2017-07-05

Received: 2016-05-18

Revision Accepted: 2016-09-12

Crosschecked: 2017-06-16

Cited: 0

Clicked: 2984

Citations:  Bibtex RefMan EndNote GB/T7714


Hao Zhang


-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2017 Vol.18 No.7 P.626-634


Functional characterization of the promoter of carbonyl reductase 1 gene in porcine endometrial cells

Author(s):  Ai-ling Zhang, Xian-yue Sun, Qi Yin, Jian-hua Zeng, Zhe Zhang, Jia-qi Li, Hao Zhang

Affiliation(s):  Guangdong Development Center of Applied Ecology and Ecological Engineering in Universities, Biology and Food Engineering Institute, Guangdong University of Education, Guangzhou 510310, China; more

Corresponding email(s):   zhangmeixial@163.com, zhanghao@scau.edu.cn

Key Words:  Pig, Carbonyl reductase 1 (CBR1), Promoter, NF-κ, B, Endometrium

Ai-ling Zhang, Xian-yue Sun, Qi Yin, Jian-hua Zeng, Zhe Zhang, Jia-qi Li, Hao Zhang. Functional characterization of the promoter of carbonyl reductase 1 gene in porcine endometrial cells[J]. Journal of Zhejiang University Science B, 2017, 18(7): 626-634.

@article{title="Functional characterization of the promoter of carbonyl reductase 1 gene in porcine endometrial cells",
author="Ai-ling Zhang, Xian-yue Sun, Qi Yin, Jian-hua Zeng, Zhe Zhang, Jia-qi Li, Hao Zhang",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Functional characterization of the promoter of carbonyl reductase 1 gene in porcine endometrial cells
%A Ai-ling Zhang
%A Xian-yue Sun
%A Qi Yin
%A Jian-hua Zeng
%A Zhe Zhang
%A Jia-qi Li
%A Hao Zhang
%J Journal of Zhejiang University SCIENCE B
%V 18
%N 7
%P 626-634
%@ 1673-1581
%D 2017
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1600225

T1 - Functional characterization of the promoter of carbonyl reductase 1 gene in porcine endometrial cells
A1 - Ai-ling Zhang
A1 - Xian-yue Sun
A1 - Qi Yin
A1 - Jian-hua Zeng
A1 - Zhe Zhang
A1 - Jia-qi Li
A1 - Hao Zhang
J0 - Journal of Zhejiang University Science B
VL - 18
IS - 7
SP - 626
EP - 634
%@ 1673-1581
Y1 - 2017
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1600225

Prostaglandins (PGs) play a critical role in porcine reproduction, of which prostaglandin E2 (PGE2) and prostaglandin F2b1; (PGF2b1;) exert antiluteolytic and luteolysis actions, respectively. As a rate-limiting enzyme, bonyl reductase 1 (CbR1)%29&ck%5b%5D=abstract&ck%5b%5D=keyword'>b>carbonyl reductase 1 (CbR1) catalyzes the conversion of PGE2 to PGF2b1;. A high ratio of PGE2:PGF2b1; is beneficial to the establishment and maintenance of porcine pregnancy. PG is essential for the establishment of pregnancy which resembles the proinflammatory response and nuclear factor bA;b (bA%29&ck%5b%5D=abstract&ck%5b%5D=keyword'>b>NF-bA;b) is involved in the process. bioinformatic analysis has shown that bA%29&ck%5b%5D=abstract&ck%5b%5D=keyword'>b>NF-bA;b is a possible factor bound to two cis-regulatory elements in CbR1 b%5D=abstract&ck%5b%5D=keyword'>b>promoter. In this study, we cloned the 2997 bp (−2875/+122) of the b%5D=abstract&ck%5b%5D=keyword'>b>promoter, and constructed six 5'-deleted dual-luciferase reporter recombinant vectors. In endometrial cells, the region of P2 (−1640/+7) exhibited the greatest transcriptional activity at driving luciferase expression, but not significantly different from that of P1 (−2089/+7). The activity of P1, P2, and P3 (−1019/+7) was highly significantly higher than that of others (P<0.01), suggesting that two positive regulatory elements were likely present in the regions of −1640/−1019 and −1019/−647. The results also showed that the −1640/−647 region was indispensable for the b%5D=abstract&ck%5b%5D=keyword'>b>promoter. The results of chromatin immunoprecipitation (ChIP) demonstrated that the bA%29&ck%5b%5D=abstract&ck%5b%5D=keyword'>b>NF-bA;b subunit p65 binds to one site around −1545/−1531. Using four reference genes, we found that the over-expression of p65 enhanced the expression of CbR1 (P<0.05) in porcine endometrial epithelial cells, while knockdown of the p65 did not down-regulate the CbR1 expression. These results indicated that bA%29&ck%5b%5D=abstract&ck%5b%5D=keyword'>b>NF-bA;b (p65) could bind to the special element of CbR1 gene b%5D=abstract&ck%5b%5D=keyword'>b>promoter in porcine endometrial epithelial cells in vitro. The binding site of bA%29&ck%5b%5D=abstract&ck%5b%5D=keyword'>b>NF-bA;b was a positive regulator for the CbR1 gene b%5D=abstract&ck%5b%5D=keyword'>b>promoter, but was not necessary for the basic expression.


创新点:发现CBR1基因启动子在猪子宫内膜受到炎性因子核转录因子kappa B(NF-κB)成员p65调控。p65对该启动子具有正向调节作用,但是对于CBR1基因的表达并不是必需的。

关键词:猪;CBR1基因;启动子;核转录因子kappa B(NF-κB);子宫内膜

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Ackerman, W.E., Summerfield, T.L.S., Vandre, D.D., et al., 2008. Nuclear factor-kappa B regulates inducible prostaglandin E synthase expression in human amnion mesenchymal cells. Biol. Reprod., 78(1):68-76.

[2]Bazer, F.W., Thatcher, W.W., Matinat-Botte, F., et al., 1991. Composition of uterine flushings from Large White and prolific Chinese Meishan gilts. Reprod. Fertil. Dev., 3(1):51-60.

[3]Blitek, A., Ziecik, A.J., 2004. Prostaglandins F and E2 secretion by porcine epithelial and stromal endometrial cells on different days of the oestrous cycle. Reprod. Domest. Anim., 39(5):340-346.

[4]Blitek, A., Mendrzycka, A.U., Bieganska, M.K., et al., 2007. Effect of steroids on basal and LH-stimulated prostaglandins F and E2 release and cyclooxygenase-2 expression in cultured porcine endometrial stromal cells. Reprod. Biol., 7(1):73-88.

[5]Christenson, L.K., Farley, D.B., Anderson, L.H., et al., 1994. Luteal maintenance during early pregnancy in the pig: role for prostaglandin E2. Prostaglandins, 47(1):61-75.

[6]Frasor, J., Weaver, A.E., Pradhan, M., et al., 2008. Synergistic up-regulation of prostaglandin E synthase expression in breast cancer cells by 17β-estradiol and proinflammatory cytokines. Endocrinology, 149(12):6272-6279.

[7]Gadsby, J.E., Lovdal, J.A., Britt, J.H., et al., 1993. Prostaglandin F2α receptor concentrations in corpora lutea of cycling, pregnant, and pseudopregnant pigs. Biol. Reprod., 49(3):604-608.

[8]Geisert, R.D., Yelich, J.V., 1997. Regulation of conceptus development and attachment in pigs. J. Reprod. Fertil. Suppl., 52:133-149.

[9]Ghosh, D., Sawicki, M., Pletnev, V., et al., 2001. Porcine carbonyl reductase. structural basis for a functional monomer in short chain dehydrogenases/reductases. J. Biol. Chem., 276(21):18457-18463.

[10]Ghosh, S., May, M.J., Kopp, E.B., 1998. NF-κB and Rel proteins: evolutionarily conserved mediators of immune responses. Annu. Rev. Immunol., 16:225-260.

[11]Guo, C., Wang, W., Liu, C., et al., 2014. Induction of PGF2α synthesis by cortisol through GR dependent induction of CBR1 in human amnion fibroblasts. Endocrinology, 155(8):3017-3024.

[12]Inoue, J., Gohda, J., Akiyama, T., et al., 2007. NF-κB activation in development and progression of cancer. Cancer Sci., 98(3):268-274.

[13]Kalabus, J.L., Cheng, Q., Blanco, J.G., 2012. MicroRNAs differentially regulate carbonyl reductase 1 (CBR1) gene expression dependent on the allele status of the common polymorphic variant rs9024. PLoS ONE, 7(11):e48622.

[14]King, A.E., Collins, F., Klonisch, T., et al., 2010. An additive interaction between the NFκB and estrogen receptor signalling pathways in human endometrial epithelial cells. Hum. Reprod., 25(2):510-518.


[16]Laird, S.M., Tuckerman, E.M., Cork, B.A., et al., 2000. Expression of nuclear factor kappa B in human endometrium; role in the control of interleukin 6 and leukaemia inhibitory factor production. Mol. Hum. Reprod., 6(1):34-40.

[17]Lakhman, S.S., Chen, X., Gonzalez-Covarrubias, V., et al., 2007. Functional characterization of the promoter of human carbonyl reductase 1 (CBR1). Role of XRE elements in mediating the induction of CBR1 by ligands of the aryl hydrocarbon receptor. Mol. Pharmacol., 72(3):734-743.

[18]Lindström, T.M., Bennett, P.R., 2005. The role of nuclear factor kappa B in human labour. Reproduction, 130(5):569-581.

[19]Mathew, D.J., Sellner, E.M., Green, J.C., et al., 2011. Uterine progesterone receptor expression, conceptus development, and ovarian function in pigs treated with RU 486 during early pregnancy. Biol. Reprod., 84(1):130-139.

[20]Miura, T., Taketomi, A., Nishinaka, T., et al., 2013. Regulation of human carbonyl reductase 1 (CBR1, SDR21C1) gene by transcription factor Nrf2. Chem. Biol. Interact., 202(1-3):126-135.

[21]Pavlová, S., Klucska, K., Vašíček, D., et al., 2011. Transcription factor NF-κB (p50/p50, p65/p65) controls porcine ovarian cells functions. Anim. Reprod. Sci., 128(1-4):73-84.

[22]Ross, J.W., Ashworth, M.D., Mathew, D., et al., 2010. Activation of the transcription factor, nuclear factor kappa-B, during the estrous cycle and early pregnancy in the pig. Reprod. Biol. Endocrinol., 8:39.

[23]Schieber, A., Frank, R.W., Ghisla, S., 1992. Purification and properties of prostaglandin 9-ketoreductase from pig and human kidney. Identity with human carbonyl reductase. Eur. J. Biochem., 206(2):491-502.

[24]Shen, J., Zhou, C., Zhu, S., et al., 2014. Comparative transcriptome analysis reveals early pregnancy-specific genes expressed in peripheral blood of pregnant sows. PLoS ONE, 9(12):e114036.

[25]Sugino, N., Karube-Harada, A., Taketani, T., et al., 2004. Withdrawal of ovarian steroids stimulates prostaglandin F2α production through nuclear factor-κB activation via oxygen radicals in human endometrial stromal cells: potential relevance to menstruation. J. Reprod. Dev., 50(2):215-225.

[26]Tanaka, M., Ohno, S., Adachi, S., et al., 1992. Pig testicular 20β-hydroxysteroid dehydrogenase exhibits carbonyl reductase-like structure and activity. cDNA cloning of pig testicular 20β-hydroxysteroid dehydrogenase. J. Biol. Chem., 267(19):13451-13455.

[27]Telleria, C.M., Goyeneche, A.A., Stocco, C.O., et al., 2004. Involvement of nuclear factor kappa B in the regulation of rat luteal function: potential roles as survival factor and inhibitor of 20α-hydroxysteroid dehydrogenase. J. Mol. Endocrinol., 32(2):365-383.

[28]Waclawik, A., Ziecik, A.J., 2007. Differential expression of prostaglandin (PG) synthesis enzymes in conceptus during peri-implantation period and endometrial expression of carbonyl reductase/PG 9-ketoreductase in the pig. J. Endocrinol., 194(3):499-510.

[29]Waclawik, A., Jabbour, H.N., Blitek, A., et al., 2009. Estradiol-17β, prostaglandin E2 (PGE2), and the PGE2 receptor are involved in PGE2 positive feedback loop in the porcine endometrium. Endocrinology, 150(8):3823-3832.

[30]Wang, S., Li, J., Zhang, A., et al., 2011. Selection of reference genes for studies of porcine endometrial gene expression on gestational day 12. Biochem. Biophys. Res. Commun., 408(2):265-268.

[31]Weems, C.W., Weems, Y.S., Randel, R.D., 2006. Prostaglandins and reproduction in female farm animals. Vet. J., 171(2):206-228.

[32]Zhang, H., Wang, S., Liu, M., et al., 2013. Differential gene expression in the endometrium on gestation day 12 provides insight into sow prolificacy. BMC Genomics, 14(1):45.

[33]Zhang, Z., Paria, B.C., Davis, D.L., 1991. Pig endometrial cells in primary culture: morphology, secretion of prostaglandins and proteins, and effects of pregnancy. J. Anim. Sci., 69(7):3005-3015.

[34]Ziecik, A.J., 2002. Old, new and the newest concepts of inhibition of luteolysis during early pregnancy in pig. Domest. Anim. Endocrinol., 23(1-2):265-275.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE