Expression and role of c-myc protooncogene in murine preimplantation embryonic development

Journal of Assisted Reproduction and Genetics - The present study was conducted to investigate the expression and possible role of the c-mycprotooncogene in preimplantation embryos by using...     

端粒酶逆转录酶在子宫内膜异位症中的表达及意义

目的:研究端粒酶逆转录酶(hTERT)在子宫内膜异位症(EMs)在位内膜及异位内膜组织中的表达,探讨hTERT在EMs发生过程中的作用。方法:应用免疫组化SP法检测54例EMs患者及15例正常子宫内膜中hTERT的表达情况,应用显微镜及相关软件评估hTERT在EMs患者在位内膜及异位内膜组织中的表达,以及其随着月经周期变化的表达情况。分析子宫内膜异位病灶组织中hTERT表达与CA125、月经周期、r-AFs分期及年龄之间的相关关系。结果:EMs异位内膜病灶处hTERT表达水平显著高于在位内膜、正常子宫内膜(P0.05);EMs在位内膜显著高于正常子宫内膜(P0.05)。正常内膜和EMs在位内膜中,月经周期变化对hTERT的表达无明显影响。EMs病灶处hTERT的表达水平与血清中CA125水平呈正相关(r=0.367,P0.05),而与月经周期、r-AFs分期和年龄呈无明显相关性。结论:hTERT在EMs中异常表达,表明hTERT在EMs的发生过程中起着一定的作用。     

Gene expression of adhesion molecules and matrix metalloproteinases in endometriosis.

Various types of cell adhesion molecules and matrix metalloproteinases (MMPs) seem to play an important role in the invasion process of endometriosis; however, limited investigation has focused on their gene expression in human peritoneal endometriotic lesions. A total of 63 endometriotic tissues were surgically obtained from 35 women with endometriosis, which included 43 pigmented and 20 non-pigmented lesions. Gene expression levels of E-cadherin, alpha- and beta-catenin, MMP-2, MMP-9 and membrane-type 1 (MT1)-MMP in these endometriotic lesions were compared with those in normal eutopic endometrium obtained from 12 women without endometriosis. MMP-2, MMP-9 and MT1-MMP mRNA expression in pigmented lesions was significantly higher than that in normal endometrium (p < 0.05), whereas E-cadherin, alpha- and beta-catenin mRNA expression was not suppressed in endometriotic lesions. There was a close correlation between MMP-2 or MT1-MMP and E-cadherin, alpha- or beta-catenin gene expression in 63 endometriotic tissues examined (p < 0.01). Immunohistochemical expression of E-cadherin, alpha- and beta-catenin in glandular epithelial cells was positive not only for all of seven cases with normal eutopic endometrium but also for 9 of 11 with ovarian endometriosis. MMP expression in ectopic endometrium was much greater than that in eutopic endometrium. These results suggest that endometriotic tissues expressing MMPs might be invasive and simultaneously possess cell-to-cell adhesion property in pelvic peritoneal foci.     

卵巢子宫内膜异位囊肿中PTTG、bFGF和c-myc的表达及意义

目的:探讨PTTG、bFGF和c-myc在卵巢子宫内膜异位囊肿的表达及意义。方法:用免疫组化法检测60例卵巢子宫内膜异位囊肿患者异位内膜、在位内膜及34例正常子宫内膜组织中PTTG、bFGF及c-myc的表达水平。结果:在卵巢子宫内膜异位囊肿患者异位内膜中,PTTG、bFGF及c-myc的阳性表达率分别是80%、76.7%及66.7%,其阳性率明显高于在位内膜(P<0.05)及正常子宫内膜组织,在位内膜亦高于正常对照组内膜(P<0.05)。等级相关分析PTTG、bFGF及c-myc呈正相关关系。结论:卵巢子宫内膜异位囊肿中PTTG表达明显高于正常子宫内膜组织,可能是通过上调bFGF和c-myc蛋白表达在卵巢子宫内膜异位囊肿的发生发展中发挥了作用。     

DNA microarray analysis of gene expression markers of endometriosis.

OBJECTIVE: To use DNA microarray technology to examine differential gene expression in uterine endometrium versus endometriosis implants. DESIGN: Pilot study. SETTING: Volunteers in an academic research environment. PATIENT(S): Premenopausal women scheduled for surgery for suspected endometriosis. INTERVENTION(S): Surgical excision of endometriosis tissue and uterine endometrial biopsy. MAIN OUTCOME MEASURE(S): Gene expression. RESULT(S): The expression of eight genes from a total of 4,133 genes on the DNA microarray was increased in endometriosis implants compared with uterine endometrium. The eight genes were beta-actin, alpha-2 actin, vimentin, 40S ribosomal protein S23, Ig-lambda light chain, Ig germline H chain G-E-A region gamma-2 constant region gene, major histocompatibility complex class 1,C, and complement component 1 S subcomponent. CONCLUSION(S): The data demonstrate that the DNA microarray is an effective tool for the identification of differentially expressed genes between uterine and ectopic endometrium; further study of the genes identified herein will expand our understanding of the nature of endometriosis and assist in the eventual development of new treatments for endometriosis.     

Chorionic gonadotrophin and c-myc expression in growing and growth-inhibited (intermediate) trophoblasts.

FEG-3 cells are a clonal line of human choriocarcinoma and resemble villous cytotrophoblasts which are the stem cells for the syncytiotrophoblast in the placenta. FEG-3 cells synthesize and secrete the alpha subunit of human chorionic gonadotrophin (hCG). Treatment of FEG-3 cells with the chemotherapeutic drug (1 microM) methotrexate (MTX) results in an increase in nuclear diameter. Cell division is blocked and a decrease in c-myc mRNA levels in observed. The effects on cell growth and c-myc mRNA expression are reversible, and cells treated with MTX for 48 h retain their proliferative potential. Assessment of placental hormone gene expression reveals that a member of the human growth hormone gene family is expressed at extremely low levels and is unaffected by MTX treatment. Alpha and beta chorionic gonadotrophin (hCG) levels are increased by MTX treatment, but levels decrease following removal of MTX. In contrast to hCG in FEG-3 cells, non-trophoblastic or ectopic production of alpha hCG in human cervical carcinoma cells is inhibited by MTX treatment. These data indicate that MTX will induce morphological and biochemical changes in FEG-3 cells. They reveal an inverse relationship between c-myc and hCG RNA expression, and suggest different mechanisms govern trophoblast versus non-trophoblast production of alpha hCG.     

Increased expression of endoglin in the eutopic endometrium of women with endometriosis.

OBJECTIVE: To compare the angiogenic activities of endothelial cells in the eutopic endometrium of women with and without endometriosis. DESIGN: Vessels with active angiogenesis were identified using the monoclonal antibody to endoglin. SETTING: University department of obstetrics and gynecology. PATIENT(S): Twenty women with histologically confirmed endometriosis after laparotomy or laparoscopy. Women with carcinoma in situ of uterine cervix, but no evidence of endometriosis (n = 20), served as control subjects. INTERVENTION(S): Formalin-fixed, paraffin-embedded archival tissues were sectioned and stained. MAIN OUTCOME MEASURE(S): Number of vessels stained with monoclonal antibody to endoglin. RESULT(S): For all menstrual phases, the mean number of vessels with endoglin expression was significantly greater in patients with endometriosis compared with control subjects. In each menstrual phase, a significant difference was observed only during the late secretory phase. Within the group with endometriosis, the mean numbers of vessels with endoglin expression in stages I and II were not different from the numbers in stages III and IV. CONCLUSION(S): This study shows the expression of endoglin in the eutopic endometrium of women with endometriosis is significantly increased and the increase is observed only in the late secretory phase. It is suggested from these findings that activation of angiogenesis in the eutopic endometrium might be a key factor in the pathogenesis of endometriosis.     

Immunohistochemical assessment of superoxide dismutase expression in the endometrium in endometriosis and adenomyosis.

OBJECTIVE: To determine the expression of superoxide dismutase (SOD) in the endometrium during the menstrual cycle in endometriosis and adenomyosis. DESIGN: Immunohistochemical identification of SOD in endometrial tissues using the monoclonal antibody. SETTING: Department of obstetrics and gynecology in a university hospital. PATIENT(S): The subjects were divided into three groups: 36 patients with endometriosis, 38 patients with histologically proven adenomyosis, and 47 fertile control subjects. INTERVENTION(S): Endometrium was biopsied throughout the menstrual cycle. MAIN OUTCOME MEASURE(S): Semiquantitative immunostaining (evaluation nomogram) score for endometrial cells. RESULT(S): The analyses revealed phase-dependent changes in the expression of SODs in the glandular and surface epithelia during the menstrual cycle in fertile controls. Specifically, the expression of copper, zinc SOD was weakest in the early and midproliferative phases, then gradually increased, and was most marked in the early and midsecretory phases. The expression of manganese SOD reached a peak in the late secretory phase. The expression of both SODs in endometriosis and adenomyosis was persistently higher than the control levels throughout the menstrual cycle. CONCLUSION(S): The exaggerated expression of both SODs in the endometrium throughout the menstrual cycle suggests that superoxide plays a key role in infertility in endometriosis and adenomyosis.     

Pleuroperitoneal endometriosis.

A nearly unique case is presented of pleural and peritoneal endometriosis with bloody pleural effusion and 4700 cc of bloody ascites. Theories of pathogenesis, differential diagnosis, and treatment are discussed. This represents another of the protean manifestations of endometriosis to complement the many others described in the literature.     

Sema3A及其受体在子宫内膜异位症中的表达及意义

目的 检测Sema 3A及其受体在子宫内膜异位症中的表达,探讨Sema 3A及其受体在子宫内膜异位症的发生和发展中的意义。方法 收集2013年1月1日至12月31日在中山大学附属第一医院妇科住院并行手术治疗的腹膜子宫内膜异位症（PEM）患者24例、卵巢子宫内膜异位囊肿（OEM）患者24例和结直肠子宫内膜异位症（CREM）患者20例,采用免疫组化SP法检测异位病灶腺上皮细胞中Sema 3A、Plexin A1和NRP-1的表达,并与26例非子宫内膜异位症患者在位内膜（EuE-NEM）、22例子宫内膜异位症患者在位内膜（EuE-EM）中的表达进行比较。结果 Sema 3A在EuE-EM组腺上皮细胞中的表达比EuE-NEM组腺上皮细胞中的表达高（309.09±66.61、207.69±56.02）,差异有统计学意义（P<0.01）。PEM、OEM、CREM的异位病灶腺上皮细胞之间的Sema 3A、Plexin A1、NRP-1的免疫组织化学HSCORE评分差异均无统计学意义（P>0.05）。PEM组、OEM组、CREM组中异位病灶腺上皮细胞中的Sema 3A、Plexin A1、NRP-1的表达均比内膜异位症患者组的在位内膜和非内膜异位症患者组的在位内膜腺上皮细胞中的表达高（P值均<0.05）。结论 Sema 3A及其受体在不同类型内膜异位症病灶腺上皮细胞中的表达增高,提示其高表达可能参与了子宫内膜异位症的发生发展。     

miRNA-135a对子宫内膜异位症HOXA10基因表达的调控的研究

目的:探讨miRNA-135a对子宫内膜异位症(EMs)中HOXA10基因表达的调控作用。方法:选取2013年1月到12月在天津市中心妇产科医院因EMs和单纯卵巢囊肿行腹腔镜手术的患者各80例。留取EMs患者的宫腔内膜组织(EMs在位内膜组)和异位内膜组织(EMs异位内膜组)以及单纯卵巢囊肿患者的宫腔内正常内膜组织。分别采用转染技术、实时荧光定量PCR技术检测miRNA-135a和HOXA10 mRNA的表达。结果:miRNA-135a和HOXA10 mRNA在不同月经周期表达水平不同。增殖期和分泌中期,EMs在位和异位内膜组织中miRNA-135a表达水平显著高于正常内膜,HOXA10 mRNA表达水平显著低于正常内膜,差异均有统计学意义(P0.01);EMs在位和异位内膜组织比较,差异均无统计学意义(P0.05)。经miRNA-135a、miRNA-135a抑制剂转染后,子宫内膜间质细胞中HOXA10 mRNA表达水平显著下降和升高,差异均有统计学意义(P0.01)。结论:EMs患者子宫内膜中HOXA10异常表达受miRNA-135a调控,miRNA-135a高表达抑制了HOXA10基因表达。     

Livin和Caspase-9在子宫内膜异位症中的表达

目的探讨凋亡调控因子Livin和Caspase-9在子宫内膜异位症发生中的作用及二者之间的关系。方法选取子宫内膜异位症患者30例作为研究对象，分别取其在位内膜（A组）和异位内膜（B组）进行研究。选取正常子宫内膜标本30例作为对照（C组）。采用免疫组织化学SP法检测三组组织中Livin和Caspase-9蛋白，以图像中阳性区平均灰度值为检测数据。结果①Livin在A、B组[（142．55±36．27），（141．69±26．61）]的表达较C组（113．26±10．22）升高，Caspase-9在A、B组[（120．37±23．31），（119．37±17．03）]的表达较C组（154．78±10．02）降低，差异均有统计学意义（P〈0．05），二者在A、B组间比较差异均无统计学意义（P〉0．05）；②A、B组中Livin和Caspase-9的表达存在负相关性（r1=-0．609，r2=-0．443，P1〈0．01，P2〈0．05）；③Livin在A、C组增生期[（141．63±36．69），（112．01±8．93）]和分泌期[（143．75±37．16），（115．13±12．07）]表达差异均无统计学意义（P〉0．05），Caspase-9在A组增生期（118．23±24．69）和分泌期（124．07±21．31）表达差异无统计学意义（P〉0．05），在C组增生期（150．93±3．04）较分泌期（159．18±13．20）表达降低，差异具有统计学意义（P〈0．05）。结论子宫内膜异位症患者在位内膜中Livin表达增加，Caspase-9表达减少，导致患者子宫内膜细胞抗凋亡能力增强，从而在子宫内膜异位症的发生中起到一定的作用。     

Statins inhibit monocyte chemotactic protein 1 expression in endometriosis

Statins are potent inhibitors of the endogenous mevalonate pathway. Besides inhibiting cholesterol biosynthesis, statins may also demonstrate anti-inflammatory properties. Inflammation is implicated in the attachment and invasion of endometrial cells to the peritoneal surface and growth of ectopic endometrium by inducing proliferation and angiogenesis. In this study, the effect of statins on monocyte chemotactic protein 1 (MCP-1) expression in endometriotic implants in nude mouse model and in cultured endometriotic cells was evaluated. In mouse model, simvastatin decreased MCP-1 expression in a dose-dependent manner in endometriotic implants (P < .05). Similarly, both simvastatin and mevastatin revealed a dose-dependent inhibition of MCP-1 production in cultured endometriotic cells (P < .01). This inhibitory effect of the statins on MCP-1 production was reversed by the downstream substrates of the mevalonate pathway. Moreover, statins decreased MCP-1 messenger RNA expression in cultured endometriotic cells (P < .05). In conclusion, statins exert anti-inflammatory effect in endometriotic cells and could provide a potential treatment of endometriosis in the future.     

Differential expression of claudins in human endometrium and endometriosis

Membrane proteins of the claudin superfamily are important components of cellular tight and adherens junctions. Although their exact function remains unclear, these proteins may play a role in tissue remodeling, a process which is associated with several diseases including endometriosis. In the present work we analyzed the expression of 13 members of the claudin family in the endometrium and peritoneum by microarray analysis. Real-time polymerase chain reaction and immunohistochemistry in human endometrium and peritoneal endometriotic lesions were performed for validation of the expression of claudin-1, -3, -4, -5 and -7. Diminished expression of claudin-3, -4 and -7 in ectopic endometrium was frequently observed as indicated by all three methods. In contrast to a higher expression of claudin-5 mRNA detected in bulk biopsies of ectopic endometrium, immunohistochemistry revealed no alteration of claudin-5 protein expression in glandular cells of endometriosis samples. The downregulation of various members of the claudin family may contribute to endometrial cell detachment and increase the number of cells invading pelvic organs.