子宫富于细胞平滑肌瘤中肥大细胞增加的机制研究

目的 探讨子宫富于细胞平滑肌瘤中肥大细胞增加的机制。方法 采用双重免疫组化染色法,检测30例子宫富于细胞平滑肌瘤（观察组）、30例普通型平滑肌瘤（对照组）和15例子宫平滑肌肉瘤（肉瘤组）患者瘤组织肥大细胞中增殖抗原Ki-67的表达,同时检测肥大细胞和平滑肌瘤细胞中趋化因子正常T细胞表达和分泌的活性调节蛋白（RANTES）、Eotaxin、单核细胞趋化蛋白1（MCP-1）和转化生长因子B（TGF-B）的表达。结果 3组患者子宫平滑肌瘤组织中,肥大细胞基本未见Ki-67的表达。观察组RANTES、Eotaxin和TGF-β的阳性表达率分别为78％、89％、91％,而在对照组和肉瘤组中其阳性表达率分别为60％、81％、86％和39％、44％、59％,观察组中3种趋化因子的表达均较对照组和肉瘤组明显升高,差异均有统计学意义（P〈0．01）。其中,RANTES和Eotaxin的阳性表达率均与肥大细胞数量呈正相关关系（r=0．655,0．543,P〈0．01）。而在3组瘤组织中,均未见MCP-1的表达。结论 子宫富于细胞平滑肌瘤中肥大细胞数量的增加,主要是由于肥大细胞向肿瘤局部募集所致。肥大细胞的局部募集与平滑肌瘤细胞分泌的RANTES和Eotaxin有关,而与TGF-β和MCP-1无关。     

p21~(WAF1/CIP1)和P53蛋白在子宫平滑肌肿瘤的表达和意义

目的 :探讨p2 1WAF1/CIP1和P5 3蛋白在子宫平滑肌肿瘤 (USMTs)的表达和意义。方法 :应用免疫组织化学法检测 2 0例普通型子宫平滑肌瘤 (UL) ,18例子宫平滑肌肉瘤 (LMS)及 70例交界性子宫平滑肌瘤 (BLM) :包括 4 0例富于细胞型子宫平滑肌瘤 (CL)、13例奇异型子宫平滑肌瘤 (BL)、4例核分裂活跃型子宫平滑肌瘤 (ML)、10例不典型子宫平滑肌瘤 (AL)及 3例恶性潜能未定型子宫平滑肌瘤 (STUMP)的p2 1WAF1/CIP1和P5 3蛋白的表达。结果 :LMS组p2 1WAF1/CIP1蛋白表达阳性率明显高于UL组 (P <0 .0 1)和BLM组 (P<0 .0 1) ;LMS组P5 3蛋白表达阳性率显著高于UL组 (P <0 .0 1)和BLM组 (P <0 .0 1)。结论 :LMS的发病机制涉及P5 3基因功能的丧失 ;p2 1WAF1/CIP1在USMT细胞的增殖和灭活中可能起到不同的作用     

CCL28通过增强MMP-2活力促进滋养细胞浸润功能

目的:探究趋化因子CCL28对滋养细胞生物学行为的影响及其调控机制。方法:分离培养并鉴定原代人早孕蜕膜基质细胞(DSCs),ELISA法检测人DSCs与人正常绒毛膜滋养细胞系HTR8/Svneo的CCL28的分泌。采用噻唑兰(MTT)、Annexin V-FITC、Transwell方法检测人工合成的CCL28对HTR8/Svneo细胞的增殖、凋亡及浸润能力的影响。RT-PCR及明胶酶谱法阐明HTR8/Svneo细胞浸润功能改变的机制。结果:HTR8/Svneo细胞与蜕膜基质细胞均能分泌CCL28,两者共培养能促进CCL28的分泌(P0.01)。人工合成的CCL28不影响滋养细胞的增殖(P0.05)和凋亡(P0.05),但能增强细胞的浸润能力(P0.01),且主要通过受体CCR10介导。RT-PCR与明胶酶谱法提示,CCL28通过提高滋养细胞内MMP-2活力来增强其浸润能力。结论:CCL28通过与受体CCR10的结合,以自分泌或旁分泌的方式增强滋养细胞MMP-2活力来促进滋养细胞的浸润功能。     

Characterization of mast cells according to their content of tryptase and chymase in normal and neoplastic human uterine cervix

Abstract. Cabanillas-Saez A, Schalper JA, Nicovani SM, Rudolph MI. Characterization of mast cells according to their content of tryptase and chymase in normal and neoplastic human uterine cervix.
Mast cells (MC) have been associated with diverse human cancers. The primary function of these cells is to store and release a number of biologically active mediators, including the serine proteases tryptase and chymase. These proteases have been closely related with angiogenesis and tumor invasion, two critical steps during tumor progression. In the present work we analyzed the presence of MC in human uterine cervix from both normal and neoplastic tissues by using metachromatic, immunohistochemical, and enzymohistochemical staining. Tryptase-positive (MC_T)– and tryptase/chymase-positive (MC_TC)–mast cells were found in both normal and neoplastic tissues. The phenotype predominantly expressed in normal tissues as well as in benign and malignant lesions of the uterine cervix was the MC_T. The total number of MC remained constant through the different stages of malignant transformation (cervical intraepithelial neoplasia grade 1–3) but a significant increase in the invasive carcinoma (IC) group was observed, this increase being mainly due to the MC_T phenotype. Furthermore, we detected abundant MC_T but not MC_TC infiltrating tumors in sections of IC. Regarding the potent angiogenic properties described for tryptase, these findings suggest that in advanced stages of malignancy the significant number of MC_T distributed within the cervical tissues could provide an effective mechanism to create the abundantly vascularized microenvironment required for tumor cells to proliferate and disseminate.     

早孕期人绒毛趋化因子CCL2及其受体CCR2的表达

目的:分析人早孕绒毛组织趋化因子CCL2及其受体CCR2的转录和翻译表达。方法:收集人早孕期胎盘组织30例,分离出绒毛组织,采用半定量RT-PCR、免疫组织化学法分析正常人早孕绒毛组织CCL2/CCR2的表达。结果:人早孕绒毛滋养细胞中等水平转录和翻译CCL2及CCR2。结论:CCL2/CCR2表达于母-胎界面的胎儿面绒毛组织,可能参与滋养细胞功能调控。     

CCR5/CCR5 ligand-induced myeloid-derived suppressor cells are related to the progression of endometriosis

Research questionImmunological disorders have been reported to promote the progression of endometriosis. Several recent studies have shown that myeloid-derived suppressor cells (MDSC) drive the progression of endometriosis. The aim of this case–control study was to test whether CCR5 and its ligands drive MDSC accumulation and play a role in the progression of endometriosis.DesignThirty-six endometriosis patients and 20 controls were recruited. All subjects underwent laparoscopy. An ELISA kit was used to define CCR5 ligands in plasma and peritoneal fluid from endometriosis patients; flow cytometry was then used to characterize CCR5⁺MDSC in peripheral blood and peritoneal fluid.ResultsData showed that endometriosis patients displayed a significantly higher production of plasma CCL3 (P = 0.046) and peritoneal fluid CCL3/5 (P = 0.042/0.036) compared with those from the uterine leiomyoma group. Furthermore, the concentrations of peritoneal fluid CCL5 were elevated in late stage patients compared with those from the uterine leiomyoma group. Accumulation of blood CCR5⁺Mo-MDSC was detected in endometriosis patients compared with those from both the ovarian dermoid cysts and uterine leiomyoma groups. Endometriosis patients also showed an elevation of CCR5⁺MDSC and CCR5⁺Mo-MDSC in peritoneal fluid samples compared with uterine leiomyoma samples. It was also found that enrichment of CCR5⁺MDSC (r = 0.6807; P < 0.0001) and CCR5⁺Mo-MDSC (r = 0.6893; P < 0.0001) were correlated with enhanced production of CCL5 in peritoneal fluid from endometriosis patients.ConclusionsThis study showed that CCR5 and its ligands could drive the progression of endometriosis by enhancing the accumulation of MDSC. These findings might produce a promising treatment that targets CCR5⁺MDSC for endometriosis patients.     

子宫交界性平滑肌瘤的病理诊断及临床特征

在子宫平滑肌瘤与肉瘤之间存在一组交界性肿瘤，包括子宫富于细胞型，奇异型，核分裂活跃型，不典型及恶性潜能未定型平滑肌瘤。本研究对原诊断为子宫富于细胞型平滑肌效４２例，进行重新诊断，同时收集其临床资料并随访。结果：４２例中，富于细胞型平滑肌瘤５例，奇异型平滑肌瘤１例，不典型平滑肌瘤７例，恶性潜能未定型平滑肌瘤２例，肉瘤１例及普通平滑肌瘤２６例。在细胞丰富的基础上，细胞异型与核分裂的关系密切；不同月经周     

CCL2通过抑制白细胞介素-24表达增强人绒毛膜滋养细胞株的细胞活力

目的探讨白细胞介素-24(interleukin-24,IL-24)对人滋养细胞活力的调节作用。方法免疫组织化学法检测IL-24及其受体(IL-20R1、IL-20R2和IL-22R1)在正常早孕期绒毛组织的表达;In-cell Western法分析重组胸腺基质淋巴细胞生成素(thymic stromal lymphopoietin,TSLP)和趋化因子CCL2对人绒毛膜滋养细胞株HTR-8/SVneo细胞中IL-24表达的影响;MTT法分析重组IL-24和CCL2对HTR-8/SVneo细胞活力的影响。结果 IL-24及其受体均在正常早孕期绒毛组织滋养细胞中强阳性表达。与对照组相比,重组CCL2体外刺激后HTR-8/SVneo细胞IL-24表达水平显著下降(P0.001)。重组IL-24处理后,HTR-8/SVneo细胞活力显著降低(P0.05或P0.001);相反,IL-24中和性抗体明显促进其细胞活力(P0.05或P0.01),重组CCL2(100 ng/mL)可体外增强HTR-8/SVneo细胞活力(P0.01),但这一作用可被重组IL-24所抑制。结论 CCL2通过抑制IL-24分泌增强人滋养细胞株HTR-8/SVneo细胞的体外活力,从而可能利于囊胚植入和胎盘发育。     

HMGI-C和HMGI(Y)基因在子宫肌瘤中的表达

目的:研究HMGI-C、HMGI(Y)基因在子宫肌瘤及子宫肌层中的表达。方法:采用Real-timePCR、Westernblot、免疫组化方法检测HMGI-C、HMGI(Y)基因在30例子宫肌瘤患者的肌瘤组织和子宫肌层组织中的表达。结果:HMGI-CmRNA及蛋白在肌瘤组织中的表达显著高于肌层组织(P<0.01)。HMGI(Y)mRNA在肌瘤组织与肌层组织中的表达无统计学差异(P>0.05),而HMGI(Y)蛋白在肌瘤组织中的表达显著高于肌层组织(P<0.05)。结论:HMGI-C、HMGI(Y)基因的异常表达与子宫肌瘤的发生有关。     

白细胞介素17诱导子宫内膜癌HEC-1-B细胞对单核细胞趋化的研究

目的:探讨白细胞介素17(IL-17)诱导人子宫内膜癌HEC-1-B细胞对单核细胞的趋化作用,及其对趋化因子CCL2、CCL5表达的影响。方法:分别采用0ng/ml(对照组)、1ng/ml、10ng/ml、100ng/ml浓度的IL-17诱导人子宫内膜癌HEC-1-B细胞,24h后收集细胞上清液置于transwell小室的下室,上室放入THP-1细胞,计数进入下室的THP-1细胞并计算趋化指数;采用荧光定量PCR方法和ELISA方法检测1ng/ml、10ng/ml、100ng/ml IL-17对HEC-1-B细胞CCL2、CCL5 mRNA基因表达以及蛋白表达水平的影响。结果:单核细胞的趋化能力与IL-17呈浓度依赖性,趋化指数在对照组及1ng/ml、10ng/ml、100ng/ml 3个实验组分别为2.05±0.17、2.93±0.22、3.53±0.28及4.57±0.37,4组的差异有统计学意义(P<0.01)。各实验组细胞CCL2及CCL5 mRNA和蛋白表达水平与对照组比较明显升高(P<0.05),表达水平均随着IL-17浓度增加而增高。结论:IL-17诱导后的子宫内膜癌细胞液可以促进单核细胞的趋化作用,其机制可能与IL-17促进HEC-1-B细胞分泌趋化因子CCL2、CCL5的表达上调有关。     

Positron emission tomography and leiomyomas: clinicopathologic analysis of 3 cases of PET scan-positive leiomyomas and literature review

INTRODUCTION: Studies have suggested that PET scans can differentiate between leiomyomas and leiomyosarcomas. Our experience, however, shows that PET scan-positive smooth muscle tumors are not necessarily malignant. CASE REPORTS: Three patients with cancer underwent PET imaging. In all three, the most worrisome finding was a PET scan-positive uterine tumor. After surgical extirpation, all three uterine tumors were found to be benign smooth muscle neoplasms. DISCUSSION: To explore the potential reason these tumors were positive on PET imaging, we performed a detailed histopathologic and immunohistochemical study of all specimens. Pathologic evaluation revealed a leiomyoma, a cellular leiomyoma, and a stromomyoma. There was no association between an increased Ki67 (proliferative) index and positivity on PET imaging. Increased vascularity, however, appeared to be a feature common to the leiomyomas that were PET-positive.     

Estrogen and progesterone receptor expression in patients with uterine smooth muscle tumors

OBJECTIVE: To investigate the expression of estrogen and progesterone receptor in uterine leiomyomas, smooth muscle tumors of uncertain malignant potential (STUMP), and leiomyosarcomas (LMS), and to assess the correlation between steroid receptor expression and clinicopathologic parameters in LMS. DESIGN: Estrogen/progesterone receptor expression was investigated by immunohistochemistry. SETTING: Department of Gynecology and Obstetrics of the University Hospital of Vienna. PATIENT(S): Twenty-six women with leiomyoma, 24 with STUMP, and 21 with LMS of the uterus. INTERVENTION(S): Formalin-fixed, paraffin-embedded tissues were sectioned and stained. MAIN OUTCOME MEASURE(S): Number of tumor cells stained. RESULT(S): Significant differences regarding the frequency of estrogen receptor expression were observed between LMS and leiomyoma and STUMP and leiomyoma (P<.05). The progesterone receptor expression did significantly differ between LMS and STUMP (P=.05), and LMS and leiomyoma (P<.05). In uterine LMS, the relationship between estrogen/progesterone receptor expression and clinicopathologic parameters did not reach statistical significance (P>.05), and neither of the markers studied revealed prognostic significance (P>.05). CONCLUSION(S): The present study observed significant differences of steroid receptor expression between uterine leiomyoma, STUMP, and LMS. Our data indicate that the progesterone receptor may be an especially useful marker to distinguish cases of malignant smooth muscle tumors in which histological features are ambiguous or borderline.     

Histamine receptors in the female reproductive system. Part I. Role of the mast cells and histamine in female reproductive system

Histamine isolated from many different tissues, acts via three types of histamine receptors: H1, H2 and H3. In peripheral tissues histamine is mainly stored in mast cells (MC). Presence of mast cells was proved also in mammals' uteri. In human uterus the majority of mast cells are located close to smooth muscle cells. It might indicate that MC plays a role in tissue remodelling during the menstrual cycle. The quantity and activity of mast cells is in connection with hormonal status of the organism. Although there are some differences, human uterine mast cells are similar to the mast cells isolated from other tissues. It is suggested that histamine is important for normal ovulation, blastocyst implantation, placental blood flow regulation, lactation and contractile activity of uterus. Histamine may also play a role in pathological processes such as pre-eclampsia or preterm delivery. The participation of mast cells and histamine in blastocyst implantation is very controversial. In W/Wv mice (without mast cells) normal implantation was observed. It denies the main role of mast cells in this process but dos not exclude histamine action. In mice the major source of histamine are uterine epithelial cells during early pregnancy. The influence of cytokines on blastocyst implantation and the role of histamine in cytokines release from the uterine mast cells are also very unclear.     

Leiomyoma of the ovary

Ovarian leiomyoma is a rarity. Usually it is not associated with any clinical symptoms. Macro- and microscopically it is indistinguishable from the uterine leiomyoma. Treatment consists of simple removal of the tumor. The possible histogenesis of this tumor is reviewed. It is concluded that the ovarian leiomyoma probably originates from the smooth muscle cells of the ovarian blood vessel or from the smooth muscle fibers near the attachment of the ovarian ligament.     

Cotyledonoid leiomyoma of the uterus: report of a case.

A 46-year-old woman presented with a pelvic mass. At the time of operation a large, exophytic, multinodular tumor extended into the peritoneal cavity and right broad ligament from a pedunculated attachment to the uterus in the region of the right cornu. On external examination the lesion had the appearance of cotyledonoid dissecting leiomyoma. On microscopic examination bulbous processes were composed of benign smooth muscle arranged in interlacing fascicles or swirls; there was focal hydropic degeneration. Significant nuclear atypia, mitotic activity, and coagulative tumor necrosis were not encountered. No intravascular involvement was present. There was no demonstrable parent leiomyoma or intramural dissecting component, and thus the case differed from previously reported cases of both cotyledonoid dissecting leiomyoma and intramural dissecting leiomyoma. This tumor represents another variation in the group of benign uterine smooth muscle tumors with unusual growth patterns.     

Ultrastructural study of cultured smooth muscle cells from uterine leiomyoma and myometrium under the influence of sex steroids

Upon the hypothesis that the differentiation or development of smooth muscle cells in both myometrium and leiomyomas may be under the regulation of sex steroids, myometrial and myoma cells were cultured in the presence of sex steroids and their morphological changes were studied ultrastructurally. The cells from the explants of both leiomyoma and myometrium at the 14th day in either control or E (10(-8) M 17-beta-estradiol)-added medium showed extremely well-developed cytoplasmic organelles with few filaments located at the marginal portion of the cytoplasm. These ultrastructural features resembled those of a dedifferentiated stage of smooth muscle cells in vitro. At the 21st day, these cells showed a moderate number of cytoplasmic filaments with dense bodies. These ultrastructural features resembled those of a redifferentiated stage of smooth muscle cells in vitro. In both E- and P (10(-7) M progesterone)-added medium, the cultured smooth muscle cells from both leiomyoma and myometrium showed an increased number of myofilaments with dense bodies even at the 14th day to the 21st day of culture in comparison to the cells either in control or estrogen-added medium. Therefore, the cultured smooth muscle cells in the medium with E and P lacked their dedifferentiated phase in vitro and/or the differentiation or maintenance of their filaments might be under the influence of both estrogen and progesterone. On the observation that the cultured smooth muscle cells from both leiomyoma and myometrium showed very similar ultrastructural changes under the influence of sex steroids, it was suggested that leiomyoma may have the same progenitor cells as myometrium.     

Monocyte chemoattractant protein-1 (MCP-1/CCL2) in experimental autoimmune orchitis

Experimental autoimmune orchitis (EAO) is characterized by an interstitial mononuclear cell infiltrate and a severe lesion of seminiferous tubules with germ cells that undergo apoptosis and sloughing. The mechanism by which immune cells migrate and extravasate in the testicular interstitium is poorly understood. The aim of this study was to detect the variations in the expression of monocyte chemoattractant protein-1 (MCP-1/CCL2) and its receptor in the testis of rats undergoing autoimmune orchitis. EAO was induced in Sprague–Dawley adult rats by active immunization with an emulsion of testicular homogenate and complete Freund adjuvant using Bordetella pertussis as co-adjuvant. Control rats injected with saline and adjuvants and normal untreated rats were also studied. By ELISA we observed a significant increase of MCP-1 in the testicular fluid (TF) and in the conditioned medium obtained from cultures of testicular macrophages of rats with EAO compared with control groups. By immunohistochemistry, an increase in MCP-1 expression was observed in mononuclear, endothelial, Leydig and peritubular cells. MCP-1 immunoreactivity was also detected in Sertoli cell cytoplasm of rats with severe orchitis. A 2-fold increase in the number of mononuclear cells that express CCR2 was also found in rats with orchitis compared with controls. In conclusion, we demonstrated in vivo that MCP-1 is highly expressed in testicular interstitial cells suggesting that this chemokine has an important role in recruiting immune cells to the testis in rats undergoing autoimmune orchitis.     

Immunohistochemical markers in differential diagnosis of endometrial stromal sarcoma and cellular leiomyoma

OBJECTIVE: Distinction of endometrial stromal sarcoma (ESS) from benign smooth muscle proliferations like cellular leiomyoma (CL) is sometimes problematic. The purpose of this study was to evaluate the potential utility of a panel of antibodies in the differential diagnosis of ESS and CL. METHODS: Using a standard streptavidin-biotin method, the expression of desmin, alpha smooth muscle actin (SMA), calponin h1, h-caldesmon, estrogen receptor (ER), progesterone receptor (PR), CD10, CD44v3, proliferating cell nuclear antigen (PCNA), and mast cells (MCs) were evaluated in 26 cases of ESS (21 low grade, 5 high grade), 25 CL (17 common CL, 8 highly CL), 25 myometria, and 25 endometria. RESULTS: Among ESS, 20 of 26, 17 of 26, 9 of 26, 12 of 26, 14 of 26, and 22 of 26 were positive for expression of desmin, SMA, calponin h1, ER, PR, and CD10, respectively, while only 2 of 26 were positive for CD44v3 and all were entirely negative for h-caldesmon. Of CL, all were positive for SMA, calponin h1, PR, and CD44v3; 24 of 25, 24 of 25, and 19 of 25 were positive for desmin, h-caldesmon, and ER, respectively, whereas 1 of 25 focally marked with antibodies to CD10. There was no significant difference of PCNA expression between ESS and CL, although the ESS cases tended to have higher values. The MC counts were significantly higher in the CL group than in the ESS group (P < 0.01). When using the cut-off value of seven MCs per HPF to distinguish ESSs from CLs, the sensitivity and specificity of this cut-off value were 92.9% and 100%, respectively. CONCLUSIONS: A panel of h-caldesmon, CD10, and CD44v3 should be used and will distinguish ESS from CL in most cases. In addition, counting the number of MCs might be useful as part of a multivariate approach to the differential diagnosis of them. But the biological function of MC and CD44v3 in these tumors is worthy of further investigation.     

Decreased serum level of macrophage inflammatory chemokine-3beta/CCL19 in preterm labor and delivery

OBJECTIVE: Chemokines are small soluble molecules which mediate leukocyte migration and may be involved in the pathophysiology of preterm labor. We aimed to determine if serum concentrations of selected chemokines are changed in preterm labor and delivery. STUDY DESIGN: A novel array-based enzyme-linked immunosorbent assay was used to quantitate serum levels of nine chemokines from a single sample: MDC/CCL22, TARC/CCL17, ITAC/CXCL11, I-309/CCL1, IP-10/CXCL10, MIP-1alpha/CCL3, -1beta/CCL4, -3alpha/CCL20 and -3beta/CCL19. Women in preterm labor who delivered (n = 17), women at preterm pregnancy not in labor (n = 13) and women in labor at term (n = 8) participated. RESULTS: In the preterm delivery group of patients, the MIP-3beta/CCL19 concentration was in mean (+/-S.D.) 70.4+/-31.7 pg/mL, which was significantly lower than that in preterm gravidas not in labor of 123+/-34 pg/mL (p < 0.001) and those in labor at term of 118+/-25.6 pg/mL (p < 0.01). The other measured chemokines did not differ significantly. CONCLUSIONS: Of a small number of examined chemokines, we were able to show that one of them, MIP-3beta/CCL19 was significantly lower in women with preterm labor and delivery. Whether or not this chemokine has a potential as biochemical marker of preterm delivery remains to be determined.