Preoperative Treatment of Uterine Leiomyomas: Clinical Findings and Expression of Transforming Growth Factor-β3 and Connective Tissue Growth Factor

Abstract

Objective: To evaluate the clinical features and the expression of transforming grwoth factor-β3 (TGF-β3) and connective tissue growth factor (CTGF) in myometrium and uterine leiomyomas after preoperative treatment with gonadotropin-releasing hormone-analogs (GnRH-a) and tibolone.

Methods: Twenty-three patients received 3.75 mg leuprolide acetate depot for 4 months. Twenty-two ptients received the same therapy plus 2.5 mg tibolone daily. Patients underwent uterine surgery after therapy. Twenty-two untreated patients underwent surgery directly. Hematologic tests, bone mineral density (BMD) measurement, and ultrasonographic evaluation of uterine volume were performed before and after treatment. Menorrhagia and pelvic pain were evaluated with a visual analog scale. Hot flushes were recorded in daily diaries. Immunohistochemical expression of TGF-β3 and CTGF in myometrium and myoma samples was evaluated semiquantitatively.

Results: After therapy, hemoglobin and iron levels similarly increased in both groups. BMD significantly decreased only in the GnRH-a group. Uterine volume similarly decreased in both groups. No patient had menorrhagia or pelvic pain at the end of therapy. The number of hot flushes increased after the first month in the GnRH-a group; in the GnRH-a plus tibolone group, it remained constant and was lower. In untreated cases, TGF-β3 and CTGF smooth muscle cell immunoexpression was lower in myometrium than in leiomyomas. After medical treatment, growth factor immunoexpression remained unchanged in myometrial samples and was reduced in leiomyomas. Endothelial cells showed strong immunopositivity, both in untreated and in treated cases.

Conclusion: This study focuses on the effects of GnRH-a and tibolone on TGF-β3 and CTGF expression in myometrium and myomas and supports the hypothesis of a pathogenetic role of these growth factors in uterine fibromatosis.

Keywords

Uterine leiomyomas GnRH-analog Tibolone TGF-β3 CTGF

Get full access to this article

View all access options for this article.

References

1. Palomba S , Pellicano M , Affinito P , Di Carlo C , Zullo F , Nappi C . Effectiveness of short-term administration of tibolone plus gonadotropin-releasing hormoen analogue on the surgical outcome of laparoscopic myomectomy. Fertil Steril 2001;75:429-433.

2. Palomba S , Affinito P , Di Carlo C , Bifulco G , Nappi C . Long term administration of tibolone plus gonadotropin-releasing hormone agonist for treatment of uterine leiomyomas: Effectiveness and effects on vasomotor symptoms, bone mass, and lipid profile. Fertil Steril 1999;72:889-895.

3. Chegini N , Ma C , Tang XM , Williams RS . Effects of GnRH analogues, “add-back” steroid therapy, antiestrogen and antiprogestins on leiomyoma and myometrial smooth muscle cell growth and transforming growth factor-beta expression. Mol Hum Reprod 2002;8:1071-1078.

4. Hulboy DL , Rudolph LA , Matrisian LM . Matrix metalloproteinases as mediators of reproductive function. Mol Hum Reprod 1997;3:27-45.

5. Ignotz RA , Massague J . Transforming growth factor-B stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem 1986;261:4337-4345.

6. Bruner KL , Rodgers WH , Gold LI , et al. Transforming growth factor beta mediates the progesterone suppression of an epithelial metalloproteinase by adjacent stroma in the human endometrium. Proc Natl Acad Sci U S A 1995;92:7362-7366.

7. Ma C , Chegini N . Regulation of matrix metalloproteinases (MMPs) and their tissue inhibitors in human myometrial smooth muscle cells by TGF-β1. Mol Hum Reprod 1999;5:950-954.

8. Derynck R , Lindquist PB , Lee A , et al. A new type of transforming growth factor-β, TGF-β3. EMBO J 1988;7:3737-3743.

9. Arici A , Sozen I . Transforming growth factor-β3 is expressed at high levels in leiomyoma where it stimulates fibronectin expression and cell proliferation. Fertil Steril 2000;73:1006-1011.

10.

10. Dou Q , Tamuzzer RW , Williams RS , Schultz GS , Chegini N . Differential expression of matrix metalloproteinases and their tissue inhibitors in leiomyomata: A mechanism for gonadotropin releasing hormone agonist-induced tumor regression. Mol Hum Reprod 1997;3:1005-1014.

11.

11. Dou Q , Zhao Y , Tarnuzzer RW , et al. Suppression of transforming growth factor-beta (TGF beta) and TGF beta receptor messenger ribonucleic acid and protein expression in leiomyometa in women receiving gonadotropin-releasing hormone agnoist therapy. J Clin Endocrinol Metab 1996;81:3222-3230.

12.

12. Pawar S , Kartha S , Tobak FG . Differential gene expression in migrating renal epithelial cells after wounding. J Cell Physiol 1995;165:556-565.

13.

13. Xie S , Sukkar MB , Issa R , Oltmanns U , Nicholson AG , Chung KF . Regulation of TGF-beta 1-induced connective tissue growth factor expression in airway smooth muscle cells. Am J Physiol Lung Cell Mol Physiol 2005;288:L68-L76.

14.

14. Brigstock DR . The connective tissue growth factor/cysteinerich61/nephroblastoma overexpressed (CCN) family. Endocrine Rev 1999;20:189-206.

15.

15. Bradham DM , Igarashi A , Potter RL , Grotendorst GR . Connective tissue growth factor: A cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10. J Cell Biol 1999;114:1285-1294.

16.

16. Igarashi A , Okochi H , Bradham DM , Grotendorst GR . Regulation of connective tissue growth factor gene expression in human skin fibroblasts during wound repair. Mol Biol Cell 1993;4:637-645.

17.

17. Igarashi A , Nashiro K , Kikuchi K , et al. Connective tissue growth factor gene expression in tissue sections from localized scleroderma, keloid, and other fibrotic skin disorders. J Invest Dermatol 1996;106:729-733.

18.

18. Hishikawa K , Oemar BS , Tanner FC , Nakaki T , Fujii T , Luscher TF . Overexpression of connective tissue growth factor gene induces apoptosis in human aortic smooth muscle cells. Circulation 1999;100:2108-2112.

19.

19. Allen JT , Knight RA , Bloor CA , Spiteri MA . Enhanced insulin like growth factor binding protein 2 (connective tissue growth factor) expression in patients with idiopathic pulmonary fibrosis and pulmonary sarcodiasis. Am J Respir Cell Mol Biol 1992;21:693-700.

20.

20. Ito Y , Aten J , Bende RJ , et al. Expression of connective tissue growth factor in human renal fibrosis. Kidney Int 1998;53:853-861.

21.

21. Igarashi A , Hayashi N , Nashiro K , Takehara K . Differential expression of connective tissue growth factor gene induces apoptosis in human aortic smooth muscle cells. Circulation 1998;25:143-148.

22.

22. Uzumcu M , Al Homsi MF , Ball DK , et al. Localization of connective tissue growth factor in human uterine tissues. Mol Hum Reprod 2000;6:1093-1098.

23.

23. Weston G , Trajstmann AC , Gargett CE , Manuelpillai U , Vollenhoven BJ , Rogers PAW . Fibroids display and anti-angiogenic pattern gene expression profile when compared with adjacent myometrium. Mol Hum Reprod 2003;9:541-549.

24.

24. Palomba S , Affintio P , Tommaselli GA , Nappi C . A clinical trial on the effects of tibolone association with gonadotropin-releasing hormone analogues for the treatment of uterine leiomyomata. Fertil Steril 1998;70:111-118.

25.

25. Wiznitzer A , Marbach M , Hazum E , Insler V , Sharoni Y , Levy J . Gonadotropin releasing hormone specific binding sites in uterine leiomyomata. Biochem Biophys Res Commun 1998;152:1326-1331.

26.

26. Chegini N , Rong H , Dou Q , Kipersztok S , Williams RS . Gonadotropin-releasing hormone (GnRH) and GnRH receptor gene expression in human myometrium and leiomyomata and the direct action of GnRH analogs on myometrial smooth muscle cells and interaction with ovarian steroids in vitro. J Clin endocrinol Metab 1996;81:3215-3221.

27.

27. Lee BS , Nowak RA . Human leiomyoma smooth muscle cells show increased expression of transforming growth factor-β3 (TGF-β3) and altered responses to the antiproliferative effects of TGF-β. J Clin Endocrinol Metab 2001;86:913-920.

28.

28. Frazier KS , Williams S , Kothapalli D , Klapper H , Grotendorst GR . Stimulation of fibroblast cell growth, matrix production, and granulation tissue formation by connective tissue growth factor. J Invest Dermatol 1996;107:404-411.

29.

29. Di Lieto A , De Falco M , Staibano S , et al. Effects of gonadotropin-releasing hormone agonists on uterine volume and vasculature and on the immunohistochemical expression of basic fibroblast growth factor (bFGF) in uterine leiomyomas. Int J Gynecol Pathol 2003;22:353-358.