| Objective: Uterine leiomyomata are the most common benign tumors of the reproductive tract, occurring in 20-25% of women during the reproductive years over the age of 30 year, and account for over 30% of hysterectomies performed annually in China. Factors that promote the initial development of leiomyomata and regulate their growth and regression remain unknow. Similar to myometrium, leiomyomata are unicellar and mainly consist of smooth muscle cells that originate from myomertrium. However, unlike myometrium, which is quiescent, identification of mitotic activities and S phase fractions indicates the growing nature of leiomyomata. The ovarian steroids have been implicated as a major promoter of leiomyoma growth. Leiomyomata rarely develop before puberty, increase in size during pregnancy, and markedly regress after menopause. But, on the other hand, it has not been possible to demonstrate in vitro evidence of the direct growth stimulatory action of steroid hormones on isolated myometrial and leiomyoma cells. This finding indicates that intermediary factors, such as growth factors or cytokines, most likely playpivotal roles in this process.Transforming groth factor- 6s (TGF- 6s ) are members of a family of polypeptide growth factors , having three diferent isoforms in mamnials: TGF- 6 K TGF- 6 2 TGF-63 and three type receptors: TGF- 6RI, 6RIL 6RHI, which are encoded by separate genes. TGF- 6s are produced and secreted by a variety of cell types. They are now considered the prototype of multifunctional cytokines. Members of this family participate in setting up the basic body plan during embryogenesis; control the formation of neural tube, limbs, cartilage, bone and sexual organs; suppress epithelial cell growth, promote wound repair, and influence important immune and endocrine functions. Alteration in the activity of these factors in humans has been implicated in fibrosis, immunosuppression, cancer and other disorders. Among the three isoforms, TGF- 6 K 63 deserve us pay attention to.Genetic evidence show that TGF- 6RI 6RH are both requaied for TGF- 6s binding and signaling transduction. They are transmembrane serine-threonine kinases, and the 6RIII is a membrane - anchored proteoglycan without a signal structure. We hypothesize that TGF- 6s may function as an autoanne and/or paracrine factor to up-regulate the accumalation of extracellar matrix and to modulate cellular replication in leiomyomata In this present study , we try to investigate the expression of TGF-61 TGF-63 and TGF-6 type I and n receptor messenger ribonucleic acid ( mRNA ) and protein in uterine leiomyoma and myometrium as well asthe relationship between TGF-Bs and the development of leiomyoma.Material and methods: 1. Tissue collection: Themyometrial and leiomyoma tissues were obtaind from the uteris of the 30 patients with leiomyoma after hysterectomy conducted for benign disease. Myometrial samples were mostly from the uterine fundus, and leiomyoma samples were the center and the periphery of each tumor. Mean age of the patiens was 46.5. The women taking hormonallyactive medications were excluded from this study. The specimens were fixed in 4% neutral formaldenhyde immediately after the hysterectomy and then embedded in paraffin. 2 The Immunohistochemistry (IHC) Test: After the deparaffinization, the blocking of endogenous peroxidase was performed in H2O2/methanol. Membrane permeabilization was achieved by treatment of the sections with proteinase K for 2 hr in humidified chamders at room temperature with the fraction of anti- TGF- 61 , TGF-B3 TGF-BRI or TGF-BRII antiserum . Binding was visualized by using a biotin-labeled Biogenex anti-rabbit antibody, using DAB as the chromogen. Sections were counterstained with hematoeylin. On the control slides, the primary antibody was replaced by PBS. 3. In Situ Hybridization Test: After deparaffinization, the blocking of endogenous peroxidase was performed in H2O2/methanol. Membrane permeabilization was achieved by treatment of the sections with proteinase K and hybridized wi... |
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