CCAAT/enhancer Binding Protein Beta Regulates Aromatase Expression Via Multiple and Novel Cis-regulatory Sequences in Uterine Leiomyoma

Overview

Journal J Clin Endocrinol Metab

Publisher Oxford University Press

Specialty Endocrinology

Date 2008 Jan 10

PMID 18182446

Citations 12

Authors

Hiroshi Ishikawa

Veysel Fenkci

Veysel Fencki

Erica E Marsh

Ping Yin

Dong Chen

You-Hong Cheng

Scott Reisterd

Zhihong Lin

Serdar E Bulun

Affiliations

Soon will be listed here.

Abstract

Context: Control of aromatase expression in uterine leiomyoma has significant clinical implications because aromatase inhibitors reduce tumor growth and associated irregular uterine bleeding. The mechanisms that regulate aromatase expression in leiomyoma are unknown.

Objectives: We previously demonstrated that the cAMP-responsive proximal promoters I.3 and II regulate aromatase expression in vivo in uterine leiomyoma tissue. Here, we investigated the cellular and molecular mechanisms responsible for promoter I.3/II usage.

Results: In smooth muscle cells isolated from leiomyoma (LSMCs), dibutyryl cAMP significantly induced aromatase mRNA and enzyme activity. Reporter constructs of promoter I.3/II deletion and site-directed mutants with selective disruption of cis-regulatory elements in the -517/-16 bp region revealed that five out of seven elements, including three CCAAT/enhancer binding protein (C/EBP) binding sites and two cAMP response elements, were essential for cAMP-induced promoter activity. EMSAs demonstrated that nuclear extracts from LSMCs contain complexes assembled on four of the five cis-elements, with C/EBP binding sites, including a novel -245/-231 bp sequence, clearly associating with C/EBPbeta. Chromatin immunoprecipitation assays revealed that C/EBPbeta binds specifically to the promoter I.3/II region in intact cells. Dibutyryl cAMP significantly induced nuclear C/EBPbeta protein levels in LSMCs in a time-dependent manner. Conversely, knockdown of C/EBPbeta dramatically suppressed cAMP-induced aromatase mRNA and enzyme activity.

Conclusions: C/EBPbeta, which binds to multiple cis-regulatory elements in promoter I.3/II, is a key factor in the transcriptional complex controlling aromatase expression in uterine leiomyoma cells. Definition of this mechanism further may assist in designing inhibitors of aromatase specific for leiomyoma tissue.

Citing Articles

The efficacy of progestins in managing pain associated with endometriosis, fibroids and pre-menstrual syndrome: a systematic review.

Allen C, Banerjee S, Karoshi M, Humaidan P, Tahmasebi F Arch Gynecol Obstet. 2025; .

PMID: 40067480 DOI: 10.1007/s00404-025-07957-0.

Lipopolysaccharide alters CEBPβ signaling and reduces estradiol production in bovine granulosa cells.

Dickson M, Sheldon I, Bromfield J CABI Agric Biosci. 2023; 3(1).

PMID: 37576606 PMC: 10419969. DOI: 10.1186/s43170-022-00133-3.

Uterine Fibroids and Progestogen Treatment: Lack of Evidence of Its Efficacy: A Review.

Donnez J J Clin Med. 2020; 9(12).

PMID: 33291422 PMC: 7762035. DOI: 10.3390/jcm9123948.

Lung metastases from benign uterine leiomyoma: does 18-FDG-PET/CT have a role to play?.

Saadeh F, Riain C, Cormack C, Gleeson N Ir J Med Sci. 2018; 188(2):619-624.

PMID: 30074141 DOI: 10.1007/s11845-018-1876-0.

Paracrine Pathways in Uterine Leiomyoma Stem Cells Involve Insulinlike Growth Factor 2 and Insulin Receptor A.

Moravek M, Yin P, Coon 5th J, Ono M, Druschitz S, Malpani S J Clin Endocrinol Metab. 2017; 102(5):1588-1595.

PMID: 28324020 PMC: 5443334. DOI: 10.1210/jc.2016-3497.

References

Zeitoun K, Bulun S . Aromatase: a key molecule in the pathophysiology of endometriosis and a therapeutic target. Fertil Steril. 1999; 72(6):961-9. DOI: 10.1016/s0015-0282(99)00393-3. View

Sumitani H, Shozu M, Segawa T, Murakami K, Yang H, Shimada K . In situ estrogen synthesized by aromatase P450 in uterine leiomyoma cells promotes cell growth probably via an autocrine/intracrine mechanism. Endocrinology. 2000; 141(10):3852-61. DOI: 10.1210/endo.141.10.7719. View

Stewart E . Uterine fibroids. Lancet. 2001; 357(9252):293-8. DOI: 10.1016/S0140-6736(00)03622-9. View

Meng L, Zhou J, Sasano H, Suzuki T, Zeitoun K, Bulun S . Tumor necrosis factor alpha and interleukin 11 secreted by malignant breast epithelial cells inhibit adipocyte differentiation by selectively down-regulating CCAAT/enhancer binding protein alpha and peroxisome proliferator-activated receptor gamma:.... Cancer Res. 2001; 61(5):2250-5. View

Zhou J, Gurates B, Yang S, Sebastian S, Bulun S . Malignant breast epithelial cells stimulate aromatase expression via promoter II in human adipose fibroblasts: an epithelial-stromal interaction in breast tumors mediated by CCAAT/enhancer binding protein beta. Cancer Res. 2001; 61(5):2328-34. View

Farquhar C, Steiner C . Hysterectomy rates in the United States 1990-1997. Obstet Gynecol. 2002; 99(2):229-34. DOI: 10.1016/s0029-7844(01)01723-9. View

Yang S, Fang Z, Suzuki T, Sasano H, Zhou J, Gurates B . Regulation of aromatase P450 expression in endometriotic and endometrial stromal cells by CCAAT/enhancer binding proteins (C/EBPs): decreased C/EBPbeta in endometriosis is associated with overexpression of aromatase. J Clin Endocrinol Metab. 2002; 87(5):2336-45. DOI: 10.1210/jcem.87.5.8486. View

Clyne C, Speed C, Zhou J, Simpson E . Liver receptor homologue-1 (LRH-1) regulates expression of aromatase in preadipocytes. J Biol Chem. 2002; 277(23):20591-7. DOI: 10.1074/jbc.M201117200. View

Shozu M, Sumitani H, Segawa T, Murakami K, Kasai T, Inoue M . Overexpression of aromatase P450 in leiomyoma tissue is driven primarily through promoter I.4 of the aromatase P450 gene (CYP19). J Clin Endocrinol Metab. 2002; 87(6):2540-8. DOI: 10.1210/jcem.87.6.8533. View

10.

Gurates B, Sebastian S, Yang S, Zhou J, Tamura M, Fang Z . WT1 and DAX-1 inhibit aromatase P450 expression in human endometrial and endometriotic stromal cells. J Clin Endocrinol Metab. 2002; 87(9):4369-77. DOI: 10.1210/jc.2002-020522. View

11.

Shozu M, Murakami K, Segawa T, Kasai T, Inoue M . Successful treatment of a symptomatic uterine leiomyoma in a perimenopausal woman with a nonsteroidal aromatase inhibitor. Fertil Steril. 2003; 79(3):628-31. DOI: 10.1016/s0015-0282(02)04761-1. View

12.

Sofi M, Young M, Papamakarios T, Simpson E, Clyne C . Role of CRE-binding protein (CREB) in aromatase expression in breast adipose. Breast Cancer Res Treat. 2003; 79(3):399-407. DOI: 10.1023/a:1024038632570. View

13.

Cheng Y, Richardson B, Hubert M, Handwerger S . Isolation and characterization of the human syncytin gene promoter. Biol Reprod. 2003; 70(3):694-701. DOI: 10.1095/biolreprod.103.023473. View

14.

Rivera J, Christopoulos S, Small D, Trifiro M . Hormonal manipulation of benign metastasizing leiomyomas: report of two cases and review of the literature. J Clin Endocrinol Metab. 2004; 89(7):3183-8. DOI: 10.1210/jc.2003-032021. View

15.

Ackerman G, Smith M, Mendelson C, MACDONALD P, Simpson E . Aromatization of androstenedione by human adipose tissue stromal cells in monolayer culture. J Clin Endocrinol Metab. 1981; 53(2):412-7. DOI: 10.1210/jcem-53-2-412. View

16.

Otubu J, Buttram V, BESCH N, BESCH P . Unconjugated steroids in leiomyomas and tumor-bearing myometrium. Am J Obstet Gynecol. 1982; 143(2):130-3. DOI: 10.1016/0002-9378(82)90640-8. View

17.

Poli V, Mancini F, Cortese R . IL-6DBP, a nuclear protein involved in interleukin-6 signal transduction, defines a new family of leucine zipper proteins related to C/EBP. Cell. 1990; 63(3):643-53. DOI: 10.1016/0092-8674(90)90459-r. View

18.

Descombes P, Schibler U . A liver-enriched transcriptional activator protein, LAP, and a transcriptional inhibitory protein, LIP, are translated from the same mRNA. Cell. 1991; 67(3):569-79. DOI: 10.1016/0092-8674(91)90531-3. View

19.

Rossi M, Chegini N, Masterson B . Presence of epidermal growth factor, platelet-derived growth factor, and their receptors in human myometrial tissue and smooth muscle cells: their action in smooth muscle cells in vitro. Endocrinology. 1992; 130(3):1716-27. DOI: 10.1210/endo.130.3.1311246. View

20.

Bulun S, Simpson E, Word R . Expression of the CYP19 gene and its product aromatase cytochrome P450 in human uterine leiomyoma tissues and cells in culture. J Clin Endocrinol Metab. 1994; 78(3):736-43. DOI: 10.1210/jcem.78.3.8126151. View