基质金属蛋白酶及其组织抑制因子在子宫内膜癌中的表达及其意义

目的　研究基质金属蛋白酶 (matrixmetalloproteinases,MMPs) 2、9及其组织抑制因子(tissueinhibitorofmetalloproteinases ,TIMPs) 1、2在子宫内膜癌中的表达 ,探讨其与子宫内膜癌浸润转移的关系。方法　应用链霉菌抗生物素蛋白 过氧化物酶免疫组织化学方法和明胶酶谱法对 37例内膜癌及 7例绝经期妇女子宫内膜组织中MMP 2、MMP 9、TIMP 1、TIMP 2蛋白及其活性进行检测。结果　MMP 2、MMP 9及TIMP 1、TIMP 2蛋白主要分布在内膜癌细胞、血管内皮细胞及绝经期子宫内膜腺上皮细胞中 ,在间质细胞中也有少量表达。内膜癌细胞中 ,MMP 2、MMP 9及TIMP 1蛋白的表达 ,病理分级为G3内膜癌的强阳性率分别为 73%、2 0 %及 6 7% ,高于G2 (13%、0及 2 7% )、G1 者 (均为 0 ,P<0 0 5 ) ;深肌层浸润内膜癌的强阳性率分别为 6 3%、16 %及 6 8% ,高于浅肌层浸润的 8%、0及 0 (P<0 0 1) ;有淋巴结转移者的强阳性率分别为 4例中 4例、4例中 3例及 4例中 4例 ,高于无淋巴结转移者的 2 5 %、0及 2 5 % (P <0 0 5 ) ;手术病理分期为Ⅲ～Ⅳ期者强阳性率分别为 5例中 5例、5例中 3例及 5例中 5例 ,高于Ⅰ～Ⅱ期者的 30 %、0及 30 % (P <0 0 5 ) ;TIMP 2蛋白在不同病理分级、肌层浸润、淋巴结转移和手术病理分期的内膜癌细     

Inhibitory effect of ginsenoside-Rb2 on invasiveness of uterine endometrial cancer cells to the basement membrane.

Ginsenoside-Rb2 derived from ginseng inhibited invasiveness to the basement membrane of endometrial cancer cell lines Ishikawa. HHUA and HEC-1-A cells. These cells dominantly expressed matrix metalloproteinase (MMP)-2 (gelatinase A) among MMPs by zymography. Ginsenoside-Rb2 suppressed the expression and activity of MMP-2, but did not alter the expression of tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 in the cells. Therefore, ginsenoside-Rb2 might inhibit invasiveness to the basement membrane via MMP-2 suppression in some endometrial cancers, and can be used as a medicine for inhibition of secondary spreading of uterine endometrial cancers.     

Endometrial tumor invasiveness is related to metalloproteinase 2 and tissue inhibitor of metalloproteinase 2 expressions

Matrix metalloproteinase (MMPs) expression has been linked to gynecological tumor aggressiveness. The objective of this study was to determine MMP-2, MMP-7, and MMP-9 and tissue inhibitors of metalloproteinases (TIMP)-1 and TIMP-2 expression in endometrial malignancies and their relation to clinical and histologic parameters. Formalin-fixed, paraffin-embedded tumor samples from 50 patients with endometrial carcinoma treated between 1999 and 2004 were stained with specific monoclonal antibodies. The tumors were grouped according to the FIGO classification. The staining results were compared to histologic and clinical data. Semiquantitative analysis of MMP and TIMP expression showed a significant difference in TIMP-2 expression according to the histologic subtype (P = 0.03) and also a trend towards a difference in MMP-9 expression (P = 0.05). MMP-2 expression increased and TIMP-2 expression fell as the histologic grade increased (P = 0.0007, P < 0.0001, respectively). MMP-2 expression correlated with lymph node metastasis (P = 0.04), while TIMP-2 expression correlated with the depth of myometrial invasion (P = 0.01), vasculolymphatic space involvement (P = 0.02), and lymph node metastasis (P = 0.0003). These results support the involvement of MMPs and TIMPs in endometrial tumor growth and progression. High MMP-2 and low TIMP-2 expression were the most potent markers of endometrial tumors with a high risk of local and distant spread.     

EGFR过表达对子宫内膜癌细胞上皮间质转化的影响

目的:探讨表皮生长因子受体(EGFR)过表达对子宫内膜癌细胞上皮间质转化(EMT)的影响。方法:选取子宫内膜癌Ishikawa及KLE细胞株,利用脂质体将携带EGFR基因的质粒转染到Ishikawa细胞株建立EGFR过表达的Ishikawa细胞株,应用RT-PCR、Western blot检测上述3种细胞中EMT相关指标E-cadherin、α-catenin、N-cadherin、Vimentin的表达量。结果:低分化、高侵袭性的KLE细胞中EGFR呈高表达,且KLE细胞中上皮标志物E-cadherin和α-catenin的蛋白表达量均低于Ishikawa细胞(P<0.01),间质标志物N-cadherin和Vimentin的蛋白表达量则高于Ishikawa细胞(P<0.01),EGFR过表达处理后,Ishikawa细胞E-cadherin、α-catenin mRNA及蛋白表达量均显著降低(P<0.01),N-cadherin、Vimentin mRNA及蛋白表达量则显著升高(P<0.01)。结论:EGFR过表达引起子宫内膜癌细胞发生上皮间质转化。     

Differential expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 protein and mRNA in epithelial ovarian tumors

OBJECTIVE: Matrix metalloproteinase-9 (MMP-9) can degrade gelatin and type IV collagen and is known to play an important role in tumor cell invasion across the basement membrane. The tissue inhibitor of metalloproteinase-1 (TIMP-1) is able to prevent activation of pro-MMP-9 and forms a 1:1 complex with the active form of MMP-9. The aim of the present study was to investigate the expression of MMP-9 and TIMP-1 in benign, borderline, and invasive epithelial ovarian tumors. MATERIALS AND METHODS: A total of 90 patients with epithelial ovarian tumor were treated at the Brigham and Women's Hospital and were used as the study population. Immunohistochemistry and in situ hybridization were performed to detect protein and mRNA expression of MMP-9 and TIMP-1. RESULTS: In the 90 epithelial ovarian tumors tested, MMP-9 expression in tumor cells was found to be significantly enhanced in serous and mucinous ovarian carcinomas compared with benign and borderline tumors. We also observed the immunostaining of MMP-9 in stromal cells of benign, borderline, and invasive epithelial ovarian tumors. Moreover, the expression levels of TIMP-1 in tumor cells were significantly higher in borderline and invasive ovarian tumors than in benign tumors. CONCLUSION: Using an in situ hybridization technique, we disclosed a direct correlation between the presence of mRNA and protein expression for both MMP-9 and TIMP-1. The present data suggest that high levels of MMP-9 protein in invasive epithelial ovarian carcinoma are strongly associated with tumor cell invasion. Enhanced expression of TIMP-1 protein in borderline and invasive tumors indicates that endogenous TIMP-1 protein may play a paradoxical role in ovarian tumor progression.     

Expression of matrix metalloproteinase-26 and tissue inhibitors of metalloproteinases TIMP-3 and -4 in benign endometrium and endometrial cancer

OBJECTIVE: Matrix metalloproteinases (MMPs) and their physiological inhibitors, the tissue inhibitors of MMPs (TIMPs), play a key role in tumor cell invasion, angiogenesis, and growth. The aim of this study was to determine the expression and cellular distribution of MMP-26, TIMP-3, and TIMP-4 in endometrial cancers and benign endometrium throughout the menstrual cycle and the correlation with tumor histological subtype, stage, and grade. METHODS: Immunohistochemical analysis using polyclonal antibodies generated against pro- and active MMP-26, and mono- and polyclonal antibodies specific to TIMP-3 and TIMP-4, respectively, was performed. RESULTS: MMP-26, TIMP-3, and TIMP-4 are expressed in endometrial carcinomas (N = 86) and benign endometrium (N = 50) from various stages of the menstrual cycle. Semi-quantitative analysis of staining intensity indicated that endometrial carcinomas expressed more MMP-26, TIMP-3, and TIMP-4 compared to benign endometrium from the postmenopausal period, but not from the secretory phase of the menstrual cycle. The highest staining intensity was associated with endometrial epithelial cells, followed by vascular endothelial cells, myometrial smooth muscle cells, and endometrial stromal cells. Increased staining intensity of MMP-26 and TIMP-3 correlated with grade III tumors and MMP-26 and TIMP-4 with the depth of myometrial invasion in tumors histologically characterized as endometrioid adenocarcinoma, clear-cell, and papillary serous carcinoma staged/graded based on FIGO criteria. CONCLUSION: MMP-26 and TIMP-4 are expressed in endometrium and endometrial carcinoma and their elevated expression and correlation with myometrial invasion suggests that MMP-26 and TIMP-4 may play a key role in endometrial tumor progression.     

Matrix metalloproteinase 2 and tissue inhibitor of metalloproteinase 2 expression is not regulated by norgestimate or norelgestromin.

Steroid hormones regulate endometrial expression of matrix metalloproteinases (MMPs) and their inhibitors. Synthetic progestins are widely used in oral contraceptives and for hormone replacement therapy. To assess whether the synthetic progestins norgestimate and its derivative norelgestromin (17-deacetylnorgestimate) modulate the expression of MMPs, Ishikawa endometrial cancer cells were separately treated with 17 beta-estradiol, 17 alpha-hydroxyprogesterone, norgestimate and norelgestromin. Culture supernatants were assayed for MMPs 2, 3 and 9, and for tissue inhibitors of MMPs (TIMP-1 and TIMP-2) by enzyme-linked immunosorbent assays (ELISAs). No marked modulation of MMP-2 and TIMP-2 expression was observed upon incubation of the cells with the synthetic progestins. By ELISA, neither MMP-3 or MMP-9 nor TIMP-1 immunoreactivity was detected. Interestingly, TIMP-2 expression was down-regulated by 17 beta-estradiol and 17 alpha-hydroxyprogesterone.     

雌激素受体亚型α对子宫内膜癌细胞HEC-1B侵袭能力调控的研究

目的：特异性下调子宫内膜癌细胞中的ERα基因，探讨E胁亚型表达在子宫内膜癌侵袭中的作用。方法：将ERa的小干扰RNA（siRNA-small interfering RNA）转染子宫内膜癌细胞HEC-1B，通过RT-PCR和Western blot证实ERα基因的有效阻断。通过transwell小室法检测下调ERa基因表达前后HEC-1B细胞的侵袭能力；应用RT-PCR检测转染前后细胞MMP-2、MMP-9、TIMP-1和TIMP-2 mRNA表达水平的变化；Western blot及明胶酶谱分别检测细胞分泌TIMP-1、TIMP-2、MMP-2和MMP-9蛋白的水平。结果：（1）将ERα-siRNA转染HEC-1B细胞后，转染效率大于90％，ERα mRNA及蛋白的表达水平均明显下调（分别为72％，67％）；（2）下调ERα基因表达后，肿瘤细胞的侵袭能力下降（P〈0．05）；在mRNA水平和蛋白水平均可检测到ERα-siRNA组细胞的MMP-2、MMP-9表达下降（P〈0．05），TIMP-1、TIMP-2表达增加（P〈0．05）。结论：使用ERα-siRNA能够有效地阻断ERa基因表达；子宫内膜癌细胞中，17β-雌二醇对MMPs／TIMPs具有调节作用，这种作用可通过ERα介导；ERα表达水平影响子宫内膜癌细胞的侵袭能力。     

Human endometrial epithelial cells modulate the activation of gelatinase a by stromal cells

Metalloproteinases (MMPs) are central effectors in endometrial physiology. Their production is tightly regulated by ovarian steroids and cytokines. Using zymography, we investigated MMP-2 production by human endometrial cells treated with estradiol-17beta + progesterone (E(2)+P) and by various key cytokines in endometrial physiology (IL-1beta, LIF, TGF-beta, and TNF-alpha). No gelatinase activity was detected in the culture media of epithelial cells. In basal conditions, stromal cells produced the pro form of MMP-2. MMP-2 production/activation was not directly affected by cytokine treatment. Interestingly, activated MMP-2 was only detected after treatment of stromal cells with culture medium from epithelial cells. Cytokine treatment of epithelial cells increased the capacity of conditioned medium to stimulate stromal cells to activate MMP-2. As the tissue inhibitor of MMP-2 (TIMP-2) is a regulator of gelatinase A activity, its concentration was measured by ELISA. TIMP-2 production by stromal cells was not affected by cytokines or by epithelial cell-conditioned medium. These results strongly suggest that regulation of stromal MMP-2 activation involves soluble factor(s) derived from the epithelial compartment.     

Up-regulation of DNA methyltransferase 3B expression in endometrial cancers

OBJECTIVE: To understand the role of epigenetic regulation in the pathogenesis of endometrial cancer, we have characterized DNA methyltransferase 3B (DNMT3B) gene expression in normal, Grade I and Grade III endometrioid cancers, and examined DNMT3B promoter activities in endometrial cancer cell lines. METHODS: DNMT3B expression was measured in normal, Grade I, and Grade III endometrioid cancer samples. Real-time PCR and Western blot analysis were performed to compare DNMT3B mRNA and protein levels. DNMT3B levels were also compared among endometrial cell lines including those for Ishikawa, KLE, AN3, RL-95, HEC-1A, and HEC-1B. DNMT3B promoter reporter plasmids were constructed. Promoter activities in well and poorly differentiated cell lines were compared by in vitro reporter gene transfection. RESULTS: DNMT3B was significantly up-regulated in both Grade I and Grade III cancers as compared to normal controls. Western blot analysis confirmed the increased DNMT3B protein expression in cancer tissues. It was also found that the well-differentiated endometrial cell line, Ishikawa, expressed lower levels of DNMT3B than the poorly differentiated KLE cells, the expression patterns similar to those observed in tumor specimens. CONCLUSION: The results suggest that DNMT3B overexpression may play a significant role in endometrial cancer development. In addition, the transfection experiments indicated that DNMT3B promoters are more active in the poorly differentiated endometrial cancer cell lines, suggesting that the in vitro assay provides a useful model for studying the DNMT3B transactivation mechanism related to tumor transformation.     

Regulation of matrix metalloproteinases and their inhibitors in uterine endometrial cells of patients with and without endometriosis

OBJECTIVE: To determine whether alterations in the secretion and regulation of matrix metalloproteinases (MMPs) and their inhibitors are present in uterine endometrial cells from endometriosis patients. STUDY DESIGN: In an in vitro study, uterine endometrial cells from 19 regularly cycling women with and 32 without endometriosis were treated with diethyl stilbestrol, promegestone (R5020), interleukin-1 (IL-1) and tumor necrosis factor a (TNF-alpha). Culture supernatants were assayed for MMPs 1, 2, 3, and 9, and for tissue inhibitors of MMP (TIMP-1 and TIMP-2) by ELISA. RESULTS: MMP-3 was secreted in high concentrations, moderate concentrations were seen for MMP-1 and MMP-2, and very low concentrations for MMP-9. Substantially more TIMP-1 than TIMP-2 was secreted. MMP-1 and MMP-3 were uniformly attenuated by R5020, while MMP-2 was not influenced by hormone treatment. MMP-3 was upregulated by TNF-alpha in all samples while IL-1 only increased secretion in cells from endometriosis patients. CONCLUSION: The upregulation of MMP-3 by IL-1 may contribute to an increased invasiveness of uterine endometrial fragments in endometriosis patients.     

EGFR过表达降低人子宫内膜癌细胞孕激素敏感性的研究

目的:检测表皮生长因子受体(EGFR)基因在子宫内膜癌孕激素敏感细胞株Ishikawa及孕激素不敏感细胞株KLE的表达,探讨EGFR基因过表达对人子宫内膜癌细胞孕激素敏感性的影响。方法:实时定量PCR法和蛋白印迹法检测Ishikawa和KLE细胞中EGFR及PR-BmRNA和蛋白的表达。将EGFR全长cDNA真核表达质粒在脂质体介导下转染至Ishikawa细胞,同时以转染空载体和未转染的Ishikawa细胞为对照,分别应用实时定量PCR检测各组细胞EGFR、PR-BmRNA表达的变化,应用蛋白印迹法检测各组细胞EGFR、PR-B蛋白表达的变化;CCK-8法观察转染EGFR基因后Ishikawa细胞对孕激素敏感性的变化。结果:(1)Ishikawa细胞中,EGFRmRNA和蛋白的表达明显低于KLE细胞(P<0.001),而PR-BmRNA和蛋白的表达则显著高于KLE细胞(P<0.001);(2)稳定转染EGFR基因后,Ishikawa细胞中EGFRmRNA和蛋白的表达水平明显高于转染空载体和未转染的Ishikawa细胞(P<0.001),而PR-BmRNA和蛋白的表达水平则显著降低;(3)10-8、10-7、10-6、10-5mol/L的MPA对未转染和转染空载体的Ishikawa细胞的抑制作用显著(P<0.05),但对稳定过表达EGFR的Ishikawa细胞无明显抑制作用(P>0.05)。结论:转染EGFR基因能有效提高Ishikawa细胞内EGFR基因的表达,但可下调PR-B基因的表达使Ishikawa细胞对MPA不敏感。     

Expression of matrix metalloproteinase-26 and tissue inhibitor of matrix metalloproteinase-3 and -4 in endometrium throughout the normal menstrual cycle and alteration in users of levonorgestrel implants who experience irregular uterine bleeding

OBJECTIVE: To determine the expression of matrix metalloproteinase (MMP-26) and tissue inhibitor of MMP (TIMP) in the endometrium of women with normal menstrual cycles compared with users of levonorgestrel implants. DESIGN: Prospective observational study. SETTING: Academic research center. PATIENT(S): Fifty patients with normal menstrual cycles who requested permanent surgical sterilization (tubal ligation) and 35 users of levonorgestrel implants. INTERVENTION(S): Endometrial biopsy. MAIN OUTCOME MEASURE(S): Expression of MMP-26, TIMP-3, and TIMP-4 by immunohistochemistry and semiquantitative analysis of staining intensity by using the H score. RESULT(S): Endometrium from women with a normal menstrual cycle and users of levonorgestrel implants expresses MMP-26, TIMP-3, and TIMP-4. These substances are present in various types of endometrial cells; expression is strongest in surface and glandular epithelial cells, followed by vascular endothelial and endometrial stromal cells. Inflammatory and immune-related cells also stained strongly for MMP-26 and TIMPs. Semiquantitative analysis of the staining intensity of endometrial epithelial and stromal cells indicated that expression of MMP-26, TIMP-3, and TIMP-4 peaks during the early to mid-luteal phase. Expression of MMP-26 is elevated in users of levonorgestrel implants who experienced irregular uterine bleeding. CONCLUSION(S): Endometrial expression of MMP-26 and TIMP-4 is present throughout the menstrual cycle and is elevated during the early to mid-luteal phase in normally cycling women. Further elevations in MMP-26 are seen in users of levonorgestrel implants who experience irregular uterine bleeding. These substances thus seem to play a role in hormonal regulation and endometrial tissue remodeling.     

基质金属蛋白酶9及其组织抑制剂1在子宫内膜异位症组织中的表达

目的探讨基质金属蛋白酶9(MMP-9)及其组织抑制剂1(TTMP-1)在子宫内膜异位症(内异症)患者异位内膜及在位内膜中的表达,及其在内异症发病中的作用。方法选取38例根据美国生育学会修订的内异症分期法,诊断为内异症患者的卵巢内膜异位囊肿标本38份、腹膜红色病变标本16份及同期在位内膜35份组织作为研究组,以及非内异症患者的子宫内膜标本20份作为对照组。采用RT-PCR半定量技术,检测上述不同组织中MMP-9mRNA及TIMP-1mRNA的表达率及表达强度。结果两组所有标本均有TIMP-1mRNA表达,部分标本有MMP-9mRNA表达。研究组中,卵巢异位囊肿及腹膜红色病变组织MMP-9mRNA表达率分别为45%及56%,表达强度分别为0·46±0·22及0·33±0·12;同期在位内膜MMP-9mRNA表达率为57%,表达强度为0·49±0·28。卵巢异位囊肿、腹膜红色病变及同期在位内膜组织TIMP-1mRNA表达强度分别为1·67±0·79、1·45±0·68及2·31±1·21,前两者与后者比较,差异有统计学意义(P<0·05)。研究组在位内膜及对照组MMP-9的表达率分别为57%及45%;表达强度分别为0·49±0·28及0·29±0·12,两组比较,差异有统计学意义(P<0·05)。研究组在位内膜及对照组TIMP-1mRNA的表达强度分别为2·31±1·21及2·40±0·89。结论内异症患者在位内膜MMP-9mRNA的表达增强,促进了内膜的异位种植。异位内膜TIMP-1mRNA的表达减弱,可促使内异症病变的发展。     

MMP-2 and TIMP-2 expression correlates with poor prognosis in cervical carcinoma--a clinicopathologic study using immunohistochemistry and mRNA in situ hybridization.

OBJECTIVE: The spread of malignant neoplasms is closely associated with matrix and basement membrane degradation, mediated by various classes of proteolytic enzymes. Matrix metalloproteinases (MMP) appear to have a key role in the sequence of events that lead to local invasion and metastasis. The present study evaluated the role of matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinases-2 (TIMP-2), and membrane-type metalloproteinase (MT1-MMP) in cervical neoplasia. METHODS: We have analyzed 49 uterine cervical squamous cell carcinomas, 10 cases of high-grade cervical intraepithelial neoplasia (CIN II-III), and 10 control cervices for the presence of MMP-2, TIMP-2, and MT1-MMP using in situ hybridization. MMP-2 protein expression was evaluated using immunohistochemistry. Results were analyzed for possible correlation with disease outcome. RESULTS: MMP-2, TIMP-2, and MT1-MMP mRNA were localized to both stromal and tumor cells. However, an intense signal for MMP-2 was detected almost exclusively in tumor cells and was uniformly absent from CIN lesions and control cervices. Conversely, intense signals for TIMP-2 and MT1-MMP were detected in both stromal and tumor cells of invasive carcinomas, more often for the former. As with MMP-2, they were absent from CIN lesions. MMP-2 protein expression was enhanced in tumor cells compared to CIN cases and controls, significantly compared to the latter (P = 0.01). The presence of both MMP-2 and TIMP-2 mRNA in tumor cells correlated with advanced stage (P = 0.003 for MMP-2, P = 0.002 for TIMP-2) and with poor survival (P = 0.003 for MMP-2, P = 0.002 for TIMP-2) in univariate analysis. In addition, their presence in tumor cells intercorrelated (P = 0.002). In multivariate survival analysis, MMP-2 presence retained its association with survival (P = 0.004), in addition to patient age (P = 0.027) and advanced stage (P = 0. 0002). CONCLUSIONS: Both MMP-2 and TIMP-2 have a key role in extracellular matrix invasion in cervical carcinoma, largely through their elaboration by tumor cells. The presence of mRNA for both proteins is interrelated and is associated with poor survival.     

Hepatocyte growth factor (HGF) induces invasion of endometrial carcinoma cell lines in vitro.

OBJECTIVES: The overall goal of this study was to investigate the role of the hepatocyte growth factor (HGF)/Met pathway in the pathophysiology of invasive endometrial carcinoma. Our objectives were (1) to examine expression of HGF and Met in surgical endometrial carcinoma specimens and endometrial carcinoma cell lines, and (2) to determine if HGF would stimulate invasion of endometrial carcinoma cell lines in vitro. METHODS: Using RT-PCR and Western immunoblotting, endometrial carcinoma specimens and the endometrial carcinoma cell lines KLE, HEC-1A, HEC-1B, and RL-95 were examined for expression of HGF and Met. A Boyden chamber invasion assay using collagen type I coated 8-microm porous membranes was then used to determine if HGF would stimulate cell invasion. Last, we assessed the capacity of endometrial stromal cells, isolated from normal human endometrium, to produce HGF as determined by an enzyme-linked immunosorbent assay and to stimulate invasion of the KLE cell line. RESULTS: All of the endometrial carcinoma tissue samples were found to express Met mRNA, and two of four samples expressed HGF mRNA. However, the endometrial carcinoma cell lines expressed only Met and not HGF mRNA. Both the endometrial carcinoma tissue specimens and the endometrial carcinoma cell lines expressed the 140-kDa Met protein. HGF induced the invasion of the KLE and HEC-1A cells through the collagen-coated membranes in a dose-dependent fashion. The optimal concentration of HGF was between 10 and 100 ng/ml. HGF (10 ng/ml) stimulated KLE invasion 1.8-fold (P < 0.05) and HEC-1A invasion 6.5-fold (P < 0.05). During exposure to endometrial stromal cell conditioned medium containing HGF as determined by ELISA, invasion of the KLE cell line was stimulated 2.5-fold (P < 0.05). CONCLUSION: These results demonstrate that HGF stimulates the invasion of endometrial carcinoma cells in vitro. Since endometrial adenocarcinoma specimens express Met, these findings suggest that the HGF/Met pathway may play a role in the invasive progression of endometrial carcinoma.     

Progestin suppresses matrix metalloproteinase production in endometrial cancer

OBJECTIVES: Endometrial carcinoma (EC) is one of the few cancers where there is a clear relationship between excessive hormone stimulation and malignant transformation. In this study we have analyzed the effects of the female sex steroids estrogen and progesterone on matrix metalloproteinases (MMP-9 and -2) production in primary EC cells and EC cell lines. MMPs are implicated in cancer invasion via mechanisms including extracellular matrix degradation and the processing of a range of molecules, including growth factors and cytokines. METHODS: Cells were isolated from biopsies collected from three cancer patients undergoing hysterectomy for grade 1 endometrial adenocarcinoma and two patients undergoing procedures unrelated to EC. These cells plus the EC cell lines Ishikawa and HEC-1A were cultured without hormones or with medroxyprogesterone acetate (MPA), estradiol (E(2)), or these hormones in combination. Gelatin and reverse zymography were used to analyze MMPs and TIMPs, respectively, in culture medium. RT-PCR was used to characterize steroid receptor expression. RESULTS: Cell lines differed from primary cells in the range and abundance of MMPs secreted. Treatment with MPA significantly reduced proMMP-9, proMMP-2, and MMP-2 release from primary EC cancer and stromal cells. Treatment with E(2) alone or MPA + E(2) had no significant effect on MMP expression. Primary EC and stromal cells also showed a loss of the progesterone B receptor isoform. CONCLUSION: EC cells retain the suppression of MMPs by progesterone, seen in normal endometrial cells. These data provide a rationale for the use of progestin therapy in the treatment of early stage grade 1 endometrial carcinomas.     

与雌、孕激素受体相关的微小RNA在Ⅰ型和Ⅱ型子宫内膜癌中的差异表达

目的 探讨与雌激素受体α(Erα)和孕激素受体(PR)表达相关的微小RNA(microRNA,miRNA)在Ⅰ型和Ⅱ型内膜癌中的差异表达.方法 通过对两种细胞系Ishikawa和KLE细胞的裸鼠移植瘤组织进行病理形态学观察、免疫组化法检测其Erα、PR和p53蛋白的表达以及活细胞计数(CCK-8)法检测雌、孕激素作用后Ishikawa和KLE细胞的生长情况,对Ishikawa和KLE细胞进行组织学分型及鉴定;对无雌、孕激素环境下培养的lshikawa和KLE细胞,用miRNA微阵列芯片筛选Ishikawa和KLE中差异表达的miRNA;用miRANDA和TargetScan软件结合芯片筛选结果,预测Ishikawa和KLE细胞中可能以ESRI(翻译产物为Erα蛋白)和PGR(翻译产物为PR蛋白)为靶基因的miRNA;用实时荧光定量PCR技术验证在体内和体外培养的lshikawa和KLE细胞中以及10例内膜癌患者癌组织标本中其表达的差异性.结果 经组织学分型及鉴定显示,lshikawa细胞来源于Ⅰ型内膜癌,KLE细胞来源于Ⅱ型内膜癌;miRNA微阵列芯片筛选出Ishikawa和KLE细胞中差异表达的miRNA共126个;可能以ESRI为靶基因的miRNA为has-miR-99a与has-miR-100,可能以PGR为靶基因的miRNA为has-miR-378与has-miR-768-3p;实时荧光定量PCR技术验证显示,has-miR-100、99a、378、768-3p在体外和体内培养的Ishikawa和KLE细胞中的差异表达与miRNA微阵列芯片筛选的结果是一致的;Ⅰ型内膜癌组织中has-miR-100的表达明显低于Ⅱ型内膜癌组织(P＜0.01).结论 has-miR-100在Ⅱ型内膜癌组织中的表达明显高于Ⅰ型内膜癌,其靶基因可能为ESR1.