促性腺激素释放激素受体，芳香化酶P450，雌、孕激素受体在子宫在位与异位内膜的表达与临床意义
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摘要
研究背景:子宫内膜异位症(Endometriosis EM)是最常见的妇科疾病之一,主要发生于生育年龄女性,近年来发病率呈逐年上升趋势,但其病因及发病机制至今未完全清楚。近年来研究表明,子宫内膜异位症是激素依赖性疾病。子宫内膜异位症的发生发展具有激素依赖性,异位组织的激素受体的表达状况直接影响子宫内膜异位症的发生和发展。目前已明确的异位组织表达的受体有雌激素受体(ER)、孕激素受体(PR)等。近年来研究发现,子宫内膜异位症患者的异位内膜组织中有促性腺激素释放激素受体(GnRHR)表达,推测GnRH可能通过其受体介导,以旁分泌或自分泌的机制作用于靶细胞,对异位内膜细胞的种植和生长直接产生作用。细胞色素芳香化酶P450(P450arom)是雌激素合成过程中的关键酶。研究表明芳香化酶可促进内异病灶局部合成雌激素,通过与雌激素受体(ER)结合促进异位病灶的生长。因此,研究子宫在位与异位内膜组织各种激素受体的表达,有助于进一步探讨性激素在子宫内膜异位症发病机制中的作用,以更有效地利用激素抑制剂治疗子宫内膜异位症。本研究采用免疫组化的方法通过比较GnRHR,P450芳香酶,雌、孕激素受体在子宫在位內膜和不同部位的异位内膜中的表达及其相关性的分析,对子宫内膜异位症发病机制进行进一步的探讨,从而为更有效地运用GnRH拮抗剂,芳香化酶抑制剂等治疗子宫内膜异位症提供临床参考。
资料与方法:收集行手术治疗的40名内异症患者在位内膜和不同部位的异位内膜组织标本共83份。按内异发生的不同部位分组如下:Ⅰ组(在位内膜组)25份;Ⅱ组(卵巢内异组)23份;Ⅲ组(阴道直肠隔内异组)11份;Ⅳ组(腹
Background: Endometriosis is a common and frequently occurring disease. The effect of its treatment is often unsatisfying because the occurrence and progression of ectopic endometrium are far from being completely understood. We conclude that it would be related to various pathegenisis by its various manifestation. Endometriosis is considered as a kind of hormone-dependent disease. Studies have demonstrated that the occurrence and development of endometriosis have hormone dependence. The expression of hormone receptors in ectopic tissues could effect the growth and implantation of endometriosis. The estrogen receptor(ER), progesterone receptor(PR) had been detected in actopic endometrium. Present studies have found that gonadotropin-releasing hormone receptor(GnRHR) express in ectopic endometrial cells and speculated whether GnRH mediated by its receptor may also exert a direct modulatory effect on endometriosis growth by autocrine-paracrine regulators. Aromatase cytothrome P450 (P450arom) is a key enzyme during the estrogen synthesis, the expression of this gene in
endometriosis suggest that ectopic endometrium may compose estrogen by paracrine secretion to anticipate the ocurrence and progression of the disease. Studies on the expression of sex hormone receptors in eutopic and ectopic endometrium will make great contribution to detecting the role of hormone in endometriosis mechanism and to treating endometriosis with hormone inhibitor better.Objective: to determine whether GnRHR, P450arom, ER and PR can be expressed in eutopic and ectopic endometrium with different location and to find the relationship among them; to detect the role of GnRHR, P450arom,ER and PR in endometriosis mechanism; to give clinic reference for using GnRHa and aromatase inhibitor better in the endometriosis treatment.Samples and Method: This study included 40 patients who had undergone operation because of endometriosis. All are at the stage of III and IV, according to the r-AFS. Their eutopic endometrium were at the proliferative phase. No hormone was used within 3 months. 83 tissues from different locations were taken from the patients, which were clinically and pathologically confirmed. Tissues from different locations had been formalin-fixed and pareffin-embedded. Normal endometrium were obtained from 20 fertile women who had endometrial curettage before put in IUD. Meanwhile, the proliferative phase of the tissues were confirmed by pathological assay. Groups: Endometriosis group: eutopic
endometrium, 25 cases; ovarian endometriosis tissues, 23 cases; vagina-rectum endometriosis tissues, 11 cases; peritoneum endometriosis tissues, 8 case; uterine serosa endometriosis tissues, 16 cases. Control group: normal endometrium, 20 cases. Two-steps immunohistochemically. Staining intensity was assigned with a semiquantitative HSCORE standard. Statistical Analysis: Analysis of variance for measurement data; Chi-square test for enumeration data; SPSS 11.5 for analysis.Results: GnRHR could be expressed with different levels at different locations. Positive reactive substance was localized in the cytoplasm of glandular cells, and had negative reaction in the nucleolus. The tissues had high expression in glandular epithelial cells while having weak or no expression in stromal cells. P450arom expression was similar to GnRHR, which were localized in the cytoplasm, primarily in glandular epithelial cells. ER and PR were expressed to a higher level in the nucleolus in both the glandular and stromal cells.The expression of GnRHR in diseased endometrium showed significantly higher than that in normal endometrium(P<0.001).; Expressions of GnRHR in ovary and vagina-rectum endometriosis were no difference with those in other groups(p>0.05); Expressions of GnRHR in peritoneum and uterine serosa endometriosis were higher than those in normal endometrial tissues(P<0.05).The expression of P450arom in eutopic endometrium were
significantly higher than that in normal endometrium (P<0.001); Expressions of P450arom in ovary, vagina-rectum
资料与方法:收集行手术治疗的40名内异症患者在位内膜和不同部位的异位内膜组织标本共83份。按内异发生的不同部位分组如下:Ⅰ组(在位内膜组)25份;Ⅱ组(卵巢内异组)23份;Ⅲ组(阴道直肠隔内异组)11份;Ⅳ组(腹
Background: Endometriosis is a common and frequently occurring disease. The effect of its treatment is often unsatisfying because the occurrence and progression of ectopic endometrium are far from being completely understood. We conclude that it would be related to various pathegenisis by its various manifestation. Endometriosis is considered as a kind of hormone-dependent disease. Studies have demonstrated that the occurrence and development of endometriosis have hormone dependence. The expression of hormone receptors in ectopic tissues could effect the growth and implantation of endometriosis. The estrogen receptor(ER), progesterone receptor(PR) had been detected in actopic endometrium. Present studies have found that gonadotropin-releasing hormone receptor(GnRHR) express in ectopic endometrial cells and speculated whether GnRH mediated by its receptor may also exert a direct modulatory effect on endometriosis growth by autocrine-paracrine regulators. Aromatase cytothrome P450 (P450arom) is a key enzyme during the estrogen synthesis, the expression of this gene in
endometriosis suggest that ectopic endometrium may compose estrogen by paracrine secretion to anticipate the ocurrence and progression of the disease. Studies on the expression of sex hormone receptors in eutopic and ectopic endometrium will make great contribution to detecting the role of hormone in endometriosis mechanism and to treating endometriosis with hormone inhibitor better.Objective: to determine whether GnRHR, P450arom, ER and PR can be expressed in eutopic and ectopic endometrium with different location and to find the relationship among them; to detect the role of GnRHR, P450arom,ER and PR in endometriosis mechanism; to give clinic reference for using GnRHa and aromatase inhibitor better in the endometriosis treatment.Samples and Method: This study included 40 patients who had undergone operation because of endometriosis. All are at the stage of III and IV, according to the r-AFS. Their eutopic endometrium were at the proliferative phase. No hormone was used within 3 months. 83 tissues from different locations were taken from the patients, which were clinically and pathologically confirmed. Tissues from different locations had been formalin-fixed and pareffin-embedded. Normal endometrium were obtained from 20 fertile women who had endometrial curettage before put in IUD. Meanwhile, the proliferative phase of the tissues were confirmed by pathological assay. Groups: Endometriosis group: eutopic
endometrium, 25 cases; ovarian endometriosis tissues, 23 cases; vagina-rectum endometriosis tissues, 11 cases; peritoneum endometriosis tissues, 8 case; uterine serosa endometriosis tissues, 16 cases. Control group: normal endometrium, 20 cases. Two-steps immunohistochemically. Staining intensity was assigned with a semiquantitative HSCORE standard. Statistical Analysis: Analysis of variance for measurement data; Chi-square test for enumeration data; SPSS 11.5 for analysis.Results: GnRHR could be expressed with different levels at different locations. Positive reactive substance was localized in the cytoplasm of glandular cells, and had negative reaction in the nucleolus. The tissues had high expression in glandular epithelial cells while having weak or no expression in stromal cells. P450arom expression was similar to GnRHR, which were localized in the cytoplasm, primarily in glandular epithelial cells. ER and PR were expressed to a higher level in the nucleolus in both the glandular and stromal cells.The expression of GnRHR in diseased endometrium showed significantly higher than that in normal endometrium(P<0.001).; Expressions of GnRHR in ovary and vagina-rectum endometriosis were no difference with those in other groups(p>0.05); Expressions of GnRHR in peritoneum and uterine serosa endometriosis were higher than those in normal endometrial tissues(P<0.05).The expression of P450arom in eutopic endometrium were
significantly higher than that in normal endometrium (P<0.001); Expressions of P450arom in ovary, vagina-rectum
引文
1.石一复,主编.子宫内膜异位症.第一版.上海:上海科学技术出版社,2002.12-23.
2. Casan EM, Raga F, Bonilla-Musoles F, et al. Human oviductal gonadotropin-release hormone in fertilization, early embryonic development, and implantation[J]. J Clin Endocrinol Metab, 2000, 85(4):1377-1381.
3. Tieva A, Stattin P, Wikstrom P, et al. Gonadotropin-realeasing hormone receptor expressionin the human prostate[J]. Prostate, 2002, 47(4): 276-284.
4. Grundlker C, Gunthert AR, Millar RP, et al. Expression of gonadotropin-releasing hormone Ⅱ(GnRH-Ⅱ) receptor in human endometrial and ovarian cancer cells and effects of GnRH-Ⅱ on tumor cell proliferation[J]. J Clin Endacrinol Metab, 2002, 87(3): 1427-1430.
5. Mangia A, Tommasi S, Reshkin SJ, et al. Gonadotropin releasing hormone receptor in primary breast cancer: comparison of immunohistochemical, radioligand and Western blot analyses[J]. Oncol Rep, 2002, 9(5): 1127-1132.
6. Borroni R, Di Blasio AM, Gaffuri B, et al. Expression of GnRH receptor gene in human ectopic endometrial cells and inhibition of their proliferation by leuprolide acetate[J]. Mol Cell Endocrinology, 2000, 159(1-2): 37-43.
7. Graddy LH, Kowalski AA, Simmen FA et al. Multiple isoforms of porcine aromatase are encoded by there distinct genes. J Steroid Biochem Mol Biol, 2000, 73(1):49-52.
8. Kitawaki J, Noguchi T, Amastu T et al. Expression of aromatase cytochrome P450protein and messenger ribonucleic acid in human endometriostic and adenomyotic tissues but not in normal endometrium. Biol Reprod, 1997, 57(3):514-520
9. Bulum SE, Zeitoun KM, Takayama K at al. Estrogen biosynthesis in endometriosis: molecular basis and clinical relevance. J Mol Endocrinol, 2000, 25:35-42.
10. Jiang J, Wu RF, Wang ZH, et al. Effect of mifepristone on estrogen and progesterone receptors in human endometrial and endometriotic cells in vitro. Fetil Steril, 2002, 77(5): 995-1000.
11. Vignali M. Molecular action of GnRH analogues on ectopic endometrial cells[J], Gynecol Obstet Invest, 1998, 45 suppl 1:2-5, discussion 35.
12. Shalev E, Leung PC. Gonadotropin-releasing hormone and reproductive medicine[J]. J Obstet Gynaecol Can, 2003, 25 (2) : 98-113.
13.陈咏健,李美芝.子宫内膜细胞色素P450芳香化酶的表达研究进展.生殖医学杂志,2001,10(2):118-121.
14. Kitawaki J, Kusuki I, Koshiba H et al. Detection of aromatase cytochrome P450 in endometrial biopsy specimens as a diagnostic test for endometriosis. Fertil Steril, 1999, 72:1100-1106.
15. Takayama K, Zeitoun K, Gunby RT, et al. Treatment of severe postmenopausal endometriosis with an aromatase ingibitor. Fertil Steril, 1998, 69: 709-713.
16. Matsuzaki S, Fukaya T, Uehara S, et al. Characterization of messenger RNA expression of estrogen receptor-alpha and -beta in patients with ovarian endometriosis. Fertil Steril, 2000, 73: 1219-1225.
17.郎景和.子宫内膜异位症的研究与设想.中华妇产科杂志,2003,38:478-480.
18. Ishihara H, Kitawaki J, Kado N, et al. Gonadotropin-releasing hormone agonist and danazol normalize aromatase cytochrome P450 expression in eutopic endometrium from women with endometriosis, adenomyosis, or leimyomas. Fertile Steril, 2003, 79 Suppl 1: 735-742.
2. Casan EM, Raga F, Bonilla-Musoles F, et al. Human oviductal gonadotropin-release hormone in fertilization, early embryonic development, and implantation[J]. J Clin Endocrinol Metab, 2000, 85(4):1377-1381.
3. Tieva A, Stattin P, Wikstrom P, et al. Gonadotropin-realeasing hormone receptor expressionin the human prostate[J]. Prostate, 2002, 47(4): 276-284.
4. Grundlker C, Gunthert AR, Millar RP, et al. Expression of gonadotropin-releasing hormone Ⅱ(GnRH-Ⅱ) receptor in human endometrial and ovarian cancer cells and effects of GnRH-Ⅱ on tumor cell proliferation[J]. J Clin Endacrinol Metab, 2002, 87(3): 1427-1430.
5. Mangia A, Tommasi S, Reshkin SJ, et al. Gonadotropin releasing hormone receptor in primary breast cancer: comparison of immunohistochemical, radioligand and Western blot analyses[J]. Oncol Rep, 2002, 9(5): 1127-1132.
6. Borroni R, Di Blasio AM, Gaffuri B, et al. Expression of GnRH receptor gene in human ectopic endometrial cells and inhibition of their proliferation by leuprolide acetate[J]. Mol Cell Endocrinology, 2000, 159(1-2): 37-43.
7. Graddy LH, Kowalski AA, Simmen FA et al. Multiple isoforms of porcine aromatase are encoded by there distinct genes. J Steroid Biochem Mol Biol, 2000, 73(1):49-52.
8. Kitawaki J, Noguchi T, Amastu T et al. Expression of aromatase cytochrome P450protein and messenger ribonucleic acid in human endometriostic and adenomyotic tissues but not in normal endometrium. Biol Reprod, 1997, 57(3):514-520
9. Bulum SE, Zeitoun KM, Takayama K at al. Estrogen biosynthesis in endometriosis: molecular basis and clinical relevance. J Mol Endocrinol, 2000, 25:35-42.
10. Jiang J, Wu RF, Wang ZH, et al. Effect of mifepristone on estrogen and progesterone receptors in human endometrial and endometriotic cells in vitro. Fetil Steril, 2002, 77(5): 995-1000.
11. Vignali M. Molecular action of GnRH analogues on ectopic endometrial cells[J], Gynecol Obstet Invest, 1998, 45 suppl 1:2-5, discussion 35.
12. Shalev E, Leung PC. Gonadotropin-releasing hormone and reproductive medicine[J]. J Obstet Gynaecol Can, 2003, 25 (2) : 98-113.
13.陈咏健,李美芝.子宫内膜细胞色素P450芳香化酶的表达研究进展.生殖医学杂志,2001,10(2):118-121.
14. Kitawaki J, Kusuki I, Koshiba H et al. Detection of aromatase cytochrome P450 in endometrial biopsy specimens as a diagnostic test for endometriosis. Fertil Steril, 1999, 72:1100-1106.
15. Takayama K, Zeitoun K, Gunby RT, et al. Treatment of severe postmenopausal endometriosis with an aromatase ingibitor. Fertil Steril, 1998, 69: 709-713.
16. Matsuzaki S, Fukaya T, Uehara S, et al. Characterization of messenger RNA expression of estrogen receptor-alpha and -beta in patients with ovarian endometriosis. Fertil Steril, 2000, 73: 1219-1225.
17.郎景和.子宫内膜异位症的研究与设想.中华妇产科杂志,2003,38:478-480.
18. Ishihara H, Kitawaki J, Kado N, et al. Gonadotropin-releasing hormone agonist and danazol normalize aromatase cytochrome P450 expression in eutopic endometrium from women with endometriosis, adenomyosis, or leimyomas. Fertile Steril, 2003, 79 Suppl 1: 735-742.