Studies on leptin and leptin receptor gene expression in myometrium and uterine myomas of GnRH analogue-treated women

Aim: To test if treatment with GnRH analogue, which leads to a significant reduction in myoma volume, changes expression of leptin genes and gene coding leptin receptor isoforms in uterine myomas and in the surrounding unaltered myometrium.

Methods: Using RT-PCR, expression of leptin genes and leptin receptor genes was studied in myomas and in the surrounding myometrium in women with uterine myomas, untreated or treated with GnRH analogue. In the randomly selected cases presence of leptin protein and of leptin receptor proteins was examined also by Western blotting.

Results: Expression of leptin genes was demonstrated both in myomas and in the surrounding myometrium, and a similar pattern of expression was found for leptin receptor isoforms. The results of RT-PCR were confirmed by Western blotting, which documented the identical distribution of leptin proteins and leptin receptor proteins in studied tissues. Treatment with GnRH analogue had no effect on the expression pattern of studied genes.

Conclusion: The results of the present study on the administration of GnRH analogue to females with myomas suggest that no direct or immediate inter-relationship exists between expression of leptin genes in uterine myomas on one hand and estrogen, progesterone and leptin levels in the blood on the other. Expression seems to be of a more durable nature but factors that induce such expression remain unknown.

Leptin; Leptin receptor; Uterine myoma; GnRH analogue

[1] Andersen J.: "Growth factors and cytokines in uterine leiomyomas". Semin. Reprod. Endocrinol., 1996, 14, 269.

[2] Senturk L.M., Sozen J., Gutierrez L., Arici A.: "Interleukin 8 production and interleukin 8 receptor expression in human myometrium and leiomyoma". Am. J. Obstet Gynecol., 2001, 184, 559.

[3] Cramer S.F., Patel A.: "The frequency of uterine leiomyomas". Am. J. Clin. Pathol., 1990, 94, 435.

[4] Tsibris J.C.M., Segars J., Coppola D., Mane S., Wilbanks G.D., Obrien W.F. et al.: "Insights from gene arrays on the developmental and growth regulation of uterine leiomyomata". Fertil. Steril., 2002, 78, 114.

[5] Nowicki M., Adamkiewicz G., Brye W., Kokot F.: "The influence of lutemizing hormone-releasing hormone analog on serum leptin and body composition in women with solitary uterine myoma". Am. J. Obstet. Gynecol., 2002, 186, 340.

[6] Wizniter A., Marbach M., Hazum E., Insler V., Sharoni Y., Levy J.: "Gonadotropin-releasing specific binding sites in uterine leiomyomata". Biochem. Biophys. Res. Commun., 1988, 16, 152, 1326.

[7] Colgan T.J., Pendergast S., LeBlanc M.: "The histopathology of uterine leiomyomas following treatment with gonadotropin-releasing hormone analogues". Hum. Pathol., 1993, 24, 1073.

[8] Palomba S., Sammartino A., DiCarlo C., Affinito P., Zullo F., Nappi C.: "Effects of raloxifene treatment on uterine leiomyomas in postrnenopausal women". Fertil. Steril., 2001, 76, 38.

[9] Murphy A.A., Morales A.J., Kettel L.M., Yen S.S.C: "Regression of uterine leiomyomata to the antiprogesterone RU 486: doseresponse effect". Fertil. Steril., 1995, 64, 187.

[10] Shouzu M., Sumitani H., Segawa T., Yang H.J., Murakami K., Inoue M.: "Inhibition of in situ expression of aromatase P450 in leiomyoma of the uterus by leuprorelin acetate". J. Clin. Endocrinol. Metab., 2001, 86, 5405.

[11] Wilson E.A., Yang F., Rees D.: "Estradiol and progesterone binding in uterine leiomyomata and in normal uterine tissues". Obstet. Gynecol., 1980, 55, 20.

[12] Puuka M.J., Kontula K.K., Kauppila A.J.I., Janne 0.A., Vihko R.K.: "Estrogen receptor in human myoma tissue". Mol. Cell. Endocrinol., 1976, 6, 35.

[13] Tamaya T., Motoyama T., Ohno Y. Ide N., Tsurusak:i T., Okada H.: "Estradiol 17-b, progesterone, and 5-alpha-dihydrotestosterone receptors of uterine myometrium and myoma in the human subject". J. Steroid. Biochem., 1979, 10, 615.

[14] Buttram V., Reiter R.C.: "Uterine leiomyomata: etiology, symptomatology, and management". Fertil Steril., 1981, 36, 433.

[15] Van der Heijden 0., Chiu H.C., Park T.C., Takahashi H., LiVolsi V.A., Risinger J.I. et al.: "Allelotype analysis of uterine leiomyoma: localization of a potential tumor suppressor gene to a 4-cM region of chromosome 7 q". Mo/. Carcinog., 1998, 23, 243.

[16] Vikhlyaeva E.M., Khodzhaeva Z.S., Fantschenko N.D.: "Familial predisposition to uterine leiomyomas". Int. J. Gynacol. Obstet., 1995, 51, 127.

[17] Markowska A., Belloni A.S., Ruecinski M., Parenti A.R., Nardelli G.B., Drews K. et al.: "Leptin and leptin receptor expression in the myometrium and uterine myomas: Is leptin involved in tumor development?". Int. J. Oncol., 2005, 27, 1505.

[18] Markowska A., Rucinski M., Drews K., Malendowicz L.K.: "Further studies on leptin receptor expression in myometrium and uterine myomas". Eur. J. Gynaecol. Oncol., 2005, 26, 517.

[19] Deurenberg P., Weststrate J.A., Seidell J.C.: "Body mass index as a measure of body fatness: age and sex-specific prediction formulas". Br. J. Nutr., 1991, 65, 105.

[20] Stewart E.A.: "Uterine fibroids". Lancet, 2001, 337, 293.

[21] Rein M.S., Barbieri R.L., Friedman A.J.: "Progesterone: A critical role in pathogenesis uterine myomas". Am. J. Obstet Gynecol., 1995, 172, 14.

[22] Rein M.S., Friedman A.J., Stuart J.M., MacLanghlin D.T.: "Fibroid and myometrial steroid receptors in women treated with gonadotropin-releasing hormone agonist leuprolide acetate". Fertil. Steril., 1990, 53, 1018.

[23] Friedman A.J., Harrison-Atlas D., Barbieri R.L., Benacerraf B., Gleason R., Schiff J.: "A randomized, placebo-controlled, doubleblind study evaluating the efficacy of leuprolide acetate depot in the treatment of uterine leiomyomata". Fertil. Steril., 1989, 51,251.

[24] Wu M.H., Chuang P.C., Chen H.M., Lin C.C., T sai S.J.: "Increased leptin expression in endometriosis cells is associated with endometrial stromal cell proliferation and leptin gene up-regulation". Mol. Hum. Reprod., 2002, 8, 456.

[25] Sreenan J.J., Prayson R.A., Biscotti C.V., Thornton M.H., Easley K.A., Hart W.R.: "Histopathologic findings in 107 uterine leiomyomas treated with leuprolide acetate compared with 126 controls". Am.J. Surg. Pathol., 1996, 20, 427.

[26] Nakamura Y., Yoshimura Y.: "Treatment of uterine leiomyomas in perimenopausal women with gonadotropine- releasing hormone agonists". Clin. Obstet. Gynecol. 1993, 36, 660.

[27] Matalliotakis I.M., Koumantaki Y.G., Neonaki M.A., Goumenou A.G., Koumantakis G.E., Kuriakou D.S., Koumantakis E.E.: "Increase in serum leptin concentrations among women with endometriosis during danazol and leuprolide depot treatments". Am. J. Obstet Gynecol., 2000, 183, 58.

[28] Nowicki M., Bryc W., Kokot F.: "Hormonal regulation of appetite and body mass in patients with advanced prostate cancer with combined androgen blockade". J. Endocrinol. Invest., 2001, 24, 31.

[29] Krotkiewski M., Laudin K., Dahlgren E., Jonson P.O., Holm G.: "Effect of two modes of antiandrogen treatment on insulin sensivity and serum leptin in women with PCOS". Gynecol. Obstet. Invest., 2003, 55, 88.

[30] Witchel S.F.,A rslanian S.,L ee P.A.: "Leptin concentrations in precocious puberty or untimely puberty with and without GnRH analogue therapy". J. Pediatr. Endocrinol. Metab., 1999, 12, 839.

[31] Tommaselli G.A., Di Carlo C., Bifulco G., Di Spiezio Sardo A., Pellicano M., Nappi C.: "Serum leptin levels in patients with premenopausal syndrome treated with GnRH analogues alone and association with tibolone". Clin. Endocrinol., 2003, 59, 716.

[32] Mix H., Widjaja A., Jandl 0., Comberg M., Kaul A., Goke M. et al.: "Expression of leptin and leptin receptor isoforms in the human stomach". Gut, 2000, 47,481.

[33] Lima-Couy I., Cervero A., Bonilla-Musoles F., Pellicer A., Simon C.: "Ehdometrial leptin and leptin receptor expression in women with severe/moderate endometriosis". Mol. Human Reprod., 2004, 10, 777.

[34] Vigano P., Somigliana E., Matrone R., Dubini A., Barron C., Vignali M., Di Blasio A.M.: "Serum leptin concentrations in endometriosis". J. Clin. Endocrinol. Metab., 2002, 87, 1085.

[35] Matarese G., Alviggi C., Sanna V., Howard J.K., Lord G.M., Carravetta C. et al.: "Increased leptin levels in serum and peritoneal fluid of patients with pelvic endometriosis". J. Clin. Endocrinol. Metab., 2000, 85, 2483.

[36] Ercolani L., Florence B., Denaro M., Alexander M.: "Isolation and complete sequence of a functional human glyceraldehydes-3-phosphate dehydrogenase gene". J. Biol. Chem., 1988, 263, 15335.

[37] Brossart P., Keilholz U., Scheibenbongen C., Mohler T., Willhauck M., Hunstein W., et al.: "Detection of residual tumor cells in patients with malignant melanoma responding to immunotherapy". J. lmmunother., 1994, 15, 38

[38] Isse N., Ogawa Y., Tamura N., Masuzaki H., Mori K., Okazaki T., et al.: "Structural organization and chromosomal assignment of the human obese gene". J. Biol. Chem., 1995, 46, 27728.

[39] Chung W.K., Power-Kehoe L., Chua M., Lee R., Leibel R.L.: "Genomic structure of the human OB receptor and identification of two novel intronic microsatellites". Genome Res., 1996, 12, 1192.

[40] Bennet B.D., Solar G.P., Yuan J.Q., Mathias J., T homas Gr., Matthews W.: "A role of leptin and its cognate receptor in hematopoesis". Current Biology, 1996, 6, 1170.

[41] Cioffi J.A., Shafer A.W., Zupancic T.J., Smith-Gbur J., Mikhail A., Platika D. et al.: "Novel B219/0B. Receptor isofom应Possible role of leptin in hematopoesis and reproduction". Nature Med., 1996, 2, 585.