Increased Midkine and Estrogen Receptor-β Expression in Human Non-Small Cell Lung Cancer

Objective： Midkine （MK）, a new member of the heparin-binding growth factor family, has been found recently to have a high expression level in many tumor specimens including lung carcinoma. Estrogens may be involved in lung carcinogenesis, and estrogen receptors, mainly estrogen receptor-β （ER-β）, are present and functional in normal lung and tumor cell lines and tissues. In addition, estrogens and growth factors may promote the progression of human non-small cell lung cancer （NSCLC）. Previously, we have immunohistochemically demonstrated that MK and ER-β proteins were overexpressed in NSCLC and their expression levels were both significantly negatively correlated with the pathological classification. The purpose of this study was to further verify their expression and its correlation with NSCLC. Methods： Taking NSCLC tissues and their corresponding paraneoplastic and normal lung as research objects, we further examined the expression of MK and ER-β by meas of RT-PCR, in situ hybridization and Western blot analyses at the levels of messenger RNA （mRNA） and protein, respectively. Results： The increased MK and ER-β mRNA expression was found in NSCLC by RT-PCR and in situ hybridization analyses. Furthermore, Western blot analysis also displayed increased expression of MK and ER-β proteins in NSCLC. Finally, their correlation analysis at the levels of mRNA and protein expression in NSCLC demonstrated that MK protein level was significantly correlated to estrogen receptor-β （P〈0.01, rs=0.535）; in spite of their correlation at the mRNA level, there was no remarkable difference between MK and ER-β （P〉0.05, rs=0.178）. Conclusion： All these results in the present study confirmed that MK and ER-β were overexpressed in human NSCLC.

AP-2α对大肠癌细胞侵袭生长及ER-β表达的影响

目的:研究转录因子激活蛋白-2α基因(transcription factor activator protein-2α,AP-2α)对大肠癌SW620细胞侵袭生长的影响,并探讨其对雌激素受体β基因(estrogen receptor-β,ER-β)表达的影响及其可能的分子机制。方法:利用脂质体介导重组质粒pcDNA3.1(+)-AP-2α与空质粒pcDNA3.1(+)转染入SW620大肠癌细胞,采用基质胶黏附实验与Transwell实验检测细胞的体外黏附、侵袭生长与迁移能力,采用实时定量PCR、Western blotting、免疫荧光细胞化学检测细胞中AP-2α和ER-β在基因和蛋白水平的表达,以电泳迁移率分析(electrophoretic mobility shift assay,EMSA)方法检测转染AP-2α基因后肿瘤细胞中AP-2α的DNA结合活性及其与ER-β的结合能力。结果:AP-2α转染SW620细胞后抑制了细胞的体外黏附、侵袭与迁移能力(均P<0.05);同时细胞中ER-β基因的mRNA和蛋白表达水平显著升高(P<0.05);EMSA结果显示,AP-2α转染的SW620细胞中表达的AP-2α蛋白与ER-β基因启动子区域发生特异性结合。结论:AP-2α转染SW620细胞可显著抑制大肠癌SW620细胞体外黏附、侵袭与迁移能力,其分子机制很可能与AP-2α直接作用于ER-β基因启动子区域从而影响ER-β基因的表达有关。

Expression of voltage.gated sodium channel Nav1.5 in non.metastatic colon cancer and its associations with estrogen receptor(ER)-βexpression and clinical outcomes

Background: Voltage-gated sodium channel 1.5(Nav1.5) potentially promotes the migratory and invasive behaviors of colon cancer cells. Hitherto, the prognostic significance of Nav1.5 expression remains undetermined. The present study aimed to explore the associations of Nav1.5 expression with clinical outcomes and estrogen receptor-β(ER-β)expression in non-metastatic colon cancer patients receiving radical resection.Methods: A total of 269 consecutive patients with pathologically confirmed stages Ⅰ-Ⅲ colon cancer who underwent radical resection were selected. Nav1.5 and ER-β expression was detected by using immunohistochemistry(IHC)on tissue microarray constructed from paraffin-embedded specimens. IHC score was determined according to the percentage and intensity of positively stained cells. Statistical analysis was performed with the X-tile method, k coefficient, Chi square test or Fisher's exact test, logistic regression, log-rank test, and Cox proportional hazards models.Results: We found that Nav1.5 was commonly expressed in tumor tissues with higher mean IHC score as compared with matched tumor-adjacent normal tissues(5.1 ± 3.5 vs. 3.5 ± 2.7, P < 0.001).The high expression of Nav1.5 in colon cancer tissues was associated with high preoperative carcinoembryonic antigen level [odds ratio(OR) = 2.980;95% confidential interval(CI)1.163-7.632; P = 0.023] and high ER-β expression(OR = 2.808; 95% CI 1.243-6.343;p = 0.00 3). Log-rank test results showed that high Nav1.5 expression contributed to a low 5-year disease-free survival(DFS) rate in colon cancer patients(77.2% vs. 92.1%, P = 0.048), especially in patients with high ER-β expression tumor(76.2% vs. 91.3%, P = 0.032). Analysis with Cox proportional hazards model demonstrated that high Nav1.5 expression[hazard ratio(HR) = 2.738; 95% CI 1.100-6.819;P = 0.030] and lymph node metastasis(HR = 2.633; 95% CI 1.632-4.248; P < 0.001) were prognostic factors for unfavorable DFS in colon cancer patients.Conclusions: High expression of Nav1.5 was associated with high expression of ER-β and indicated unfavorable oncologic prognosis in patients with non-metastatic colon cancer.

phytoestrogens/insoluble fibers and colonic estrogen receptor β: randomized, double-blind, placebo-controlled study

AIM:To assess the safety and effect of the supplementation of a patented blend of dietary phytoestrogens and insoluble fibers on estrogen receptor (ER)-β and biological parameters in sporadic colonic adenomas. METHODS:A randomized, double-blind placebo-controlled trial was performed. Patients scheduled to undergo surveillance colonoscopy for previous sporadic colonic adenomas were identified, and 60 eligible patients were randomized to placebo or active dietary intervention (ADI) twice a day, for 60 d before surveillance colonoscopy. ADI was a mixture of 175 mg milk thistle extract, 20 mg secoisolariciresinol and 750 mg oat fiber extract. ER-β and ER-α expression, apoptosis and proliferation (Ki-67 LI) were assessed in colon samples. RESULTS:No adverse event related to ADI was recorded. ADI administration showed a significant increases in ER-β protein (0.822 ± 0.08 vs 0.768 ± 0.10, P = 0.04) and a general trend to an increase in ER-β LI (39.222 ± 2.69vs 37.708 ± 5.31,P = 0.06), ER-β/ER-α LI ratio (6.564 ± 10.04 vs 2.437 ± 1.53, P = 0.06), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (35.592 ± 14.97 vs 31.541 ± 11.54, P = 0.07) and Ki-67 (53.923 ± 20.91 vs 44.833 ± 10.38, P = 0.07) approximating statistical significance. A significant increase of ER-β protein (0.805 ± 0.13 vs 0.773 ± 0.13,P = 0.04), mRNA (2.278 ± 1.19vs 1.105 ± 1.07, P < 0.02) and LI (47.533 ± 15.47 vs 34.875 ± 16.67,P < 0.05) and a decrease of ER-α protein (0.423 ± 0.06vs 0.532 ± 0.11,P < 0.02) as well as a trend to increase of ER-β/ER-α protein in ADI vs placebo group were observed in patients without polyps (1.734 ± 0.20 vs 1.571 ± 0.42, P = 0.07). CONCLUSION:The role of ER-β on the control of apoptosis, and its amenability to dietary intervention, are supported in our study.

Estrogen and insulin synergistically promote endometrial cancer progression via crosstalk between their receptor signaling pathways

Objective: Despite evidence that estrogens and insulin are involved in the development and progression of many cancers, their synergistic role in endometrial carcinoma(EC) has not been analyzed yet.Methods: Here, we investigated how estrogens act synergistically with insulin to promote EC progression. Cell growth in vitro and in vivo, effects of estradiol and insulin on apoptosis and cell cycle distribution, and expression and activation of estrogen receptor(ER), insulin receptor(InsR), and key proteins in the PI3K and MAPK pathways were examined after combined stimulation with estradiol and insulin.Results: Compared to EC cells treated with estradiol or insulin alone, those treated with both estradiol and insulin exhibited stronger stimulation. Estradiol significantly induced phosphorylation of InsR-β and IRS-1, whereas insulin significantly induced phosphorylation of ER-α. In addition, treatment with both insulin and estradiol together significantly increased the expression and phosphorylation of Akt, MAPK, and ERK. Notably, InsR-β inhibition had a limited effect on estradiol-dependent proliferation,cell cycle, and apoptosis, whereas ER-α inhibition had a limited insulin-dependent effect, in EC cell lines. Insulin and estradiol individually and synergistically promoted EC xenograft growth in mice.Conclusions: Estrogen and insulin play synergistic roles in EC carcinogenesis and progression by activating InsR-β and ER-α,promoting a crosstalk between them, and thereby resulting in the activation of downstream PI3K/Akt and MAPK/ERK signaling pathways.

Correlation and Significance of Midkine and Estrogen Receptor Beta Protein Expression in Non-Small Cell Lung Cancer

OBJECTIVE Midkine (MK),a new member of the heparin-binding growth factor family,was found recently to have a highexpression level in many carcinoma specimens,including thoseof the esophagus,gall bladder,pancreas,colorectum,breast andlung.Estrogen receptor beta (ER-β),a recently cloned estrogenreceptor subtype,was also found to be highly expressed in lungtumor tissue,in contrast to a lower level of expression in normallung tissue.However,few relevant studies on these proteins havebeen published.The aims of our study were to investigate theexpression of midkine and ER-β proteins in non-small cell lungcancer (NSCLC) and to examine the relationship between theirexpression and the clinicopathologic data as well as to analyse thecorrelation of their expression in NSCLC.METHODS By immunohistochemistry,MK and ER-β were ex-amined in 24 surgically resected cases of NSCLC with their corre-sponding paraneoplastic and normal lung tissues.RESULTS MK and ER-β were overexpressed in NSCLC.Thelevels of MK and ER-β expression in NSCLC were found to be sig-nificantly negatively correlated with the pathological classification(P=0.042 and 0.021,respectively),and their expression decreasedwith a raise in the classification.Spearman's correlation analysisshowed that the correlation of their expression in NSCLC wasstrong (correlation coefficient[r_s]= 0.535,P=0.007<0.01).CONCLUSION The expression levels of MK and ER-β to someextent reflect the malignant degree of NSCLC,and their combineddetection may be of great value in early diagnosis,treatments ofpatients with NSCLC and can predict the prognoses.

Involvement of Estrogen Receptors in the Anxiolytic-Like Effect of Phytoestrogen Genistein in Rats with 12-Weeks Postovariectomy

Phytoestrogens are natural compounds found in some vegetables, and they replicate many of the physiochemical and physiological properties of estrogens, including the regulation of mood. The phytoestrogen genistein exerts anxiolytic-like effects in rats with a chronic absence of ovarian hormones, but the mechanism involved in this effect remains to be explored. The present study explored the participation of estrogen receptor-β in the anxiolytic-like effect of genistein (1.0 mg/kg, i.p., for 4 days) in Wistar rats with 12-weeks postovariectomy, considered as experimental model of post-surgical menopause. In the light/dark test, a useful tool for anxiety study and for the screening of anxiolytic drugs, genistein reduced the latency to enter and increased the time spent in the light compartment and significantly increased the frequency and time spent exploring the light compartment compared with the control group, which is considered as an anxiolytic-like effect at experimental level. All behavioral effects produced by genistein in the light/dark test were blocked by previous tamoxifen administration (5.0 mg/kg, s.c., for 6 days), a non selective antagonist for estrogen receptor-β. The effects produced by genistein or tamoxifen in this test were not related to significant changes in general motor activity evaluated in the open field test. In conclusion, the specific contribution of present investigation was identify that estrogen receptor-β is involved in the anxiolytic-like effect produced by phytoestrogen genistein in rats with a long-term absence of ovarian hormones;supporting the hypothesis that estrogen receptor-β participates in the regulation of anxiety associated with low concentration of ovarian hormones and in the anxiolytic-like effects produced by natural estrogenic compounds such as phytoestrogens.

米非司酮下调不全流产患者蜕膜雌激素受体(ER)及孕激素受体(PR)表达

目的了解不同剂量米非司酮对人工流产术后不全流产残留物中绒毛膜促性腺激素-β(β-human chorionic gonadotrophin,β-hCG)及雌激素受体(estrogen receptor,ER)、孕激素受体(progesterone receptor,PR)表达的影响。方法 60例2009年1至9月自愿在复旦大学附属妇产科医院计划生育门诊就诊的早孕人工流产术后不全流产妇女,随机分为4组:A、B、C组术前分别服用米非司酮1 400 mg(100mg,bid×7d)、700 mg(50mg,bid×7d)、350 mg(25 mg,bid×7d),D组为对照组,每组各1 5例。除D组直接行清宫术,其他组于停药后行清宫术。应用免疫组织化学EnVision法染色,半定量测定残留物中β-hCG及ER、PR水平。结果 4组β-hCG在绒毛滋养叶细胞胞质的表达差异无统计学意义(P>0.05)。ER、PR在蜕膜上皮细胞和间质细胞核中的表达,实验组较对照组表达均降低,差异均有统计学意义(P<0.05)。经两两秩和检验(修正α=0.05):ER在蜕膜上皮细胞核中的表达为A组较C、D组明显降低,差异有统计学意义(P<0.05);ER在蜕膜间质细胞核中的表达为A组较其他组明显降低,差异有统计学意义(P<0.05);A、B、C组较D组表达降低,差异有统计学意义(P<0.05)。PR在蜕膜上皮细胞核中,A、B、C组较D组表达降低,差异有统计学意义(P<0.05)。PR在蜕膜间质细胞核中的表达A组较其他组表达明显降低,差异有统计学意义(P<0.05);B组较C、D组表达降低,差异有统计学意义(P<0.05)。结论不同剂量的米非司酮对人工流产术后不全流产残留物中β-hCG的表达影响不明显。对蜕膜细胞中的ER、PR表达有抑制作用,此抑制作用可能是米非司酮治疗不全流产的作用机制之一。1 400 mg米非司酮治疗效果最显著。

ERα和ERβ在非小细胞性肺癌组织中的表达及与其临床病理的相关性研究

目的探讨雌激素受体(ER)α和β在非小细胞性肺癌组织中的表达及与其临床病理的相关性。方法选取经组织病理学检查确诊的非小细胞肺癌220例患者作为研究对象。采用免疫组化方法检测ERα和ERβ表达水平。结果两种雌激素受体表达于细胞核内,其中ERα阳性表达率为4.09%,ERβ阳性表达率为86.36%;ERα阳性表达与临床分期、病理分化程度有关:Ⅰ期患者ERα阳性表达率高于分期晚的患者(9.38%>0.00%,χ2=12.121,P<0.01);高分化患者ERα阳性表达率高于中低分化患者(14.29%>3.00%>0.00%,χ2=14.756,P<0.01)。ERβ阳性表达与患者临床分期、分化程度及病理类型有关:Ⅰ期患者ERα阳性表达率最低(78.12%,χ2=10.732,P=0.01),腺癌及高分化非小细胞性肺癌患者ERβ具有更高表达率(70.7%,χ2=7.749,P=0.02;83.3%,χ2=12.738,P<0.01)。结论 ERα和ERβ在非小细胞肺癌患者中表达情况与其临床病理特点具有相关性,提示雌激素受体表达可能在非小细胞肺癌的发生、发展中起重要作用。

育龄妇女雌激素受体β基因RsaⅠ多态性调查

目的 雌激素受体 β是一种核调节蛋白 ,其介导雌激素发挥多重作用。雌激素受体 β基因突变与某些疾病的发生有关 ,本文调查了雌激素受体基因RsaⅠ突变在女性人群中的分布。方法 采用PCR -RFLP方法检测ER β基因第 5外显子 1 0 82位核苷酸G -A突变。结果 1 0 1名女性人群中 ,ERβ基因RsaⅠ纯合突变型基因的频率为 9 9% ,杂合突变型基因的频率为 4 4 5 5 % ,野生型基因的频率为 4 5 5 5 % ,R等位基因的频率为 32 1 8,r等位基因的频率为 6 7 82 %。结论 陕西农村女性人群ERβ基因RsaⅠ多态性高于德国、新加坡妇女的突变率。需进一步研究该基因突变与疾病的相关性。

肝组织内性甾体激素受体与慢性乙型肝炎病毒复制及其抗原表达的相关性

目的探讨肝组织内雄激素受体(AR)和雌激素受体-α(ERα)含量对慢性乙型肝炎(CHB)患者血清HBV DNA载量、HBsAg水平和肝组织内HBsAg、HBcAg表达水平影响。方法 135例经肝活组织检查的CHB患者入选本研究,其中HBeAg阳性80例,HBeAg阴性55例。肝组织内AR mRNA和ERαmRNA采用反转录定量PCR检测。结果HBeAg阳性患者,血清HBV DNA载量与肝组织内AR mRNA和ERαmRNA含量均无显著相关性(r=0.132,P=0.241和r=0.130,P=0.249),血清HBsAg水平与肝组织内AR mRNA和ERαmRNA含量均呈显著正相关(r=0.355,P=0.001和r=0.246,P=0.028);肝组织内HBcAg表达水平与肝组织内AR mRNA和ERαmRNA含量均呈显著正相关(rs=0.438,P=0.000和rs=0.352,P=0.002),肝组织内HBsAg表达水平与肝组织内AR mRNA和ERαmRNA含量无显著相关性(rs=0.112,P=0.333和rs=0.024,P=0.836)。HBeAg阴性患者,血清HBV DNA载量与肝组织内AR mRNA和ERαmRNA含量均无显著相关性(r=-0.256,P=0.061和r=-0.121,P=0.385),血清HBsAg水平与肝组织内AR mRNA和ERαmRNA含量均无显著相关性(r=0.130,P=0.348和r=0.165,P=0.234);肝组织内HBcAg表达水平与肝组织内AR mRNA和ERαmRNA含量均无显著相关性(rs=-0.053,P=0.701和rs=-0.203,P=0.137),肝组织内HBsAg表达水平与肝组织内AR mRNA和ERαmRNA含量无显著相关性(rs=0.159,P=0.247和rs=0.192,P=0.160)。多因素方差分析和有序Logistic回归分析的结果显示,肝组织内AR mRNA为影响HBeAg阳性患者血清HBsAg水平和肝组织内HBcAg表达水平的独立因素。结论肝组织内AR表达水平是影响CHB患者HBV抗原表达的一个独立因素。

祛异康对子宫内膜异位症大鼠ER、β-EP的影响

目的研究中药祛异康对子宫内膜异位症模型大鼠雌激素受体(ER)和β-内啡肽(β-EP)含量的影响,探讨其治疗子宫内膜异位症的作用机理。方法将40只健康雌性大鼠随机分为4组,造模成功后,A组、B组大鼠用生理盐水灌胃,C组大鼠用丹那唑混悬液灌胃,D组大鼠用祛异康煎剂灌胃,4W后择动情期处死,用免疫组化方法检测在位、异位子宫内膜组织ER的表达及放射免疫法检测血浆中β-EP的含量。结果祛异康组与模型组比较可显著降低在位内膜、异位内膜中ER的表达(P<0.05;P<0.01);并可显著提高血浆中β-EP的含量(P<0.05)。结论祛异康治疗EMT的作用机理可能通过降调在位、异位内膜中ER的表达及提高血浆中β-EP的含量,而改善神经内分泌功能,抑制异位内膜的生长,使异位内膜萎缩,间质体积缩小,内膜腺体减少而达到治疗效果。

Cadmium-induced neurotoxicity: still much ado

Cadmium（Cd） is a highly toxic heavy metal that accumulates in living system and as such is currently one of the most important occupational and environmental pollutants. Cd reaches into the environment by anthropogenic mobilization and it is absorbed from tobacco consumption or ingestion of contaminated substances. Its extremely long biological half-life（approximately 20-30 years in humans） and low rate of excretion from the body cause cadmium storage predominantly in soft tissues（primarily, liver and kidneys） with a diversity of toxic effects such as nephrotoxicity, hepatotoxicity, endocrine and reproductive toxicities. Moreover, a Cd-dependent neurotoxicity has been also related to neurodegenerative diseases such as Alzheimer＇s and Parkinson＇s diseases, amyotrophic lateral sclerosis, and multiple sclerosis. At the cellular level, Cd affects cell proliferation, differentiation, apoptosis and other cellular activities. Among all these mechanisms, the Cd-dependent interference in DNA repair mechanisms as well as the generation of reactive oxygen species, seem to be the most important causes of its cellular toxicity. Nevertheless, there is still much to find out about its mechanisms of action and ways to reduce health risks. This article gives a brief review of the relevant mechanisms that it would be worth investigating in order to deep inside cadmium toxicity.

TGF-β stimulation of EMT programs elicits non-genomic ER-α activity and anti-estrogen resistance in breast cancer cells

Aim:Estrogen receptor-α(ER-α)activation drives the progression of luminal breast cancers.Signaling by transforming growth factor-β(TGF-β)typically opposes the actions of ER-α;it also induces epithelial-mesenchymal transition(EMT)programs that promote breast cancer dissemination,stemness and chemoresistance.The impact of EMT programs on nongenomic ER-αsignaling remains unknown and was studied herein.Methods:MCF-7 and BT474 cells were stimulated with TGF-βto induce EMT programs,at which point ER-αexpression,localization,and nongenomic interactions with receptor tyrosine kinases and MAP kinases(MAPKs)were determined.Cell sensitivity to anti-estrogens both before and after traversing the EMT program was also investigated.Results:TGF-β-stimulated MCF-7 and BT474 cells to acquire EMT phenotypes,which enhanced cytoplasmic accumulation of ER-αwithout altering its expression.Post-EMT cells exhibited:(1)elevated expression of EGFR and IGF1R,which together with Src formed cytoplasmic complexes with ER-α;(2)enhanced coupling of EGF,IGF-1 and estrogen to the activation of MAPKs;and(3)reduced sensitivity to tamoxifen,an event reversed by administration of small molecule inhibitors against the receptors for TGF-β,EGF,and IGF-1,as well as those against MAPKs.Conclusion:EMT stimulated by TGF-βpromotes anti-estrogen resistance by activating EGFR-,IGF1R-,and MAPK-dependent nongenomic ER-αsignaling.

A Meta-Analysis of Lymphatic Vessel Invasion Correlated with Pathologic Factors in Invasive Breast Cancer

Objectives: The invasive breast cancer is divided into four clinical subtypes: Luminal A-like, Luminal B-like, HER-2 positive, and triple-negative according to the expression status of estrogen receptor (ER), progesterone receptor(PR), human epidermal growth factor receptor-2 (HER-2) and Ki-67. The prognosis and treatment strategy vary with subtypes. The current studies have reported the relation between lymphatic vessel invasion (LVI) and the expression status of ER, PR, HER-2, Ki-67 in invasive breast cancer, but the results were debatable. So the meta-analysis was conducted to confirm the relation between LVI and the four factors. Methods: Literature was searched by entering the terms: breast AND (neoplasm OR cancer OR carcinoma) AND (lymphovascular OR “lymph vessel” OR “lymphatic vessel” invasion OR carcinoma embolus) AND (ER OR estrogen receptor OR PR OR progesterone receptor OR HER-2 OR human epidermal growth factor receptor-2 OR Ki-67 OR clinicopathological) in Pubmed. The merged odds ratio (OR) and 95% confidence interval (CI) were estimated using fixed-effect model. Review Manager 5.2 was used to analysis the relation between LVI and the expression status of ER, PR, HER-2, Ki-67 in invasive breast cancer respectively. The fail-safe number was used to estimate publication bias. Results: The analysis included 5 studies, LVI positive rate was significant lower in ER positive, PR positive, HER-2 negative, low Ki-67 expression group statistically. The OR and 95% CI were 0.6(0.44 - 0.81), 0.64(0.43 - 0.95), 1.52(1.03 - 2.24), 5.29(1.53 - 18.35) respectively.Conclusions:?LVI was significantly correlated with the expression status of ER, PR, HER-2 and Ki-67 in invasive breast cancer. Furthermore, LVI was consistent with poor prognostic expression status of the four factors.

Alteration of ERβ gene Rsal polymorphism may contribute to reduced fertilization rate and embryonic developmental competence

This paper aims to determine the possible role of estrogen receptor-β （ERβ） gene Rsal polymorphism on sperm fertility and early embryonic development in humans. Three groups of Chinese men were recruited： in vitro fertilization （IVF） group, including 374 couples who underwent conventional IVF; intracytoplasmic sperm injection （ICSI） group, including 294 couples who underwent an ICSI procedure using ejaculated sperm; and azoospermic group, consisting of 197 couples who underwent ICSI using either testis or epididymis sperm. Rsal polymorphism in the ERβ gene was detected by polymerase chain reaction （PCR）-restriction fragment length polymorphism technique; fertilization and high-quality embryo rates were evaluated for each group. In each group, no significant differences were found in the overall rates of fertilization and high-quality embryos among GG, AG and AA genotypes. However, the proportion of cycles possessing a satisfactory high-quality embryo rate with the AA genotype was significantly lower than that in the wild-type GG genotype from each group. These results demonstrated that sperm possessing the ERβ RsalA genotype may have reduced fertilization ability and decreased early embryonic developmental potential, which could directly or indirectly contribute to the low fertilization rate and early embryonic developmental arrest in some cases.

Correlation of expression of ER,PR and c-erbB-2 with ultrasonographic features in invasive ductal cancer of breast

Objective To analyze the relationship of the expression level of estrogen receptor(ER),progesterone receptor(PR)and human epidermal growth factor receptor-2(HER-2,c-erbB-2)of breast invasive ductal carcinoma(IDC)with ultrasonographic characteristics.Methods Totally 104 patients with IDCs confirmed pathologically were involved in this study.ER,PR and c-erbB-2 expression in the IDC specimens was determined by immunohistochemical staining technique.The correlation between the ultrasonographic features and the positive expression of ER,PR or c-erbB-2 was analyzed.Results The positive expression rate of ER and PR in the group with tumor spiculation sign and posterior acoustic attenuation was higher than that in the group without.The positive expression rate of ER differed significantly(P<0.05)while that of PR did not(P>0.05).The over-expression rate of c-erbB-2 in the group of microcalcification,sufficient blood flow and axillary lymph node metastasis was higher than that in the group of non-microcalcification,deficient blood flow or without axillary lymph node metastasis(P<0.05).The expression of ER,PR and c-erbB-2 was not related to the size of tumor(P>0.05).Conclusion The expression of ER and c-erbB-2 is closely related to the ultrasonographic characteristics of IDC,which may,to some extent,reflect the expression level of ER and c-erbB-2.

DNA甲基化抑制物对雌激素受体β在乳腺癌细胞中表达的影响及意义

目的:研究甲基化抑制物作用于乳腺癌细胞后对其ER-β表达的影响及意义。方法:抽取细胞总RNA,采用半定量逆转录-聚合酶链反应(RT-PCR)检测MCF-7与MDA-MB-231中ER-β的表达状况。在两种细胞中按作用浓度加入甲基化抑制药物5-AZA-CdR,反应一定时间后用RT-PCR的方法测定药物处理后ER-β的表达情况,分析影响及意义。结果:MCF-7的ER-β平均表达水平(0.54±0.44),药物处理后平均表达水平(0.76±0.53),较未经药物处理的细胞明显升高(P<0.05);MDA-MB-231是雌激素受体阴性的细胞株,不表达ER-β,药物处理后ER-β出现表达平均水平为(0.32±0.11),P<0.05。结论:DNA去甲基化可以诱导ER-β表达增强或再表达,对于提高乳腺癌的内分泌治疗疗效和指导新辅助化学治疗有重大意义。

Quantitative profiles of the mRNAs of ER-α and its novel variant ER-α36 in breast cancers and matched normal tissues

Objective: The novel estrogen receptor-α (ER-α) variant ER-α36 is reported to be functional in the es-trogen signaling pathway and is related to tamoxifen resistance in breast cancer. However, ER-α36 tends to be a favorable factor for survival in patients without tamoxifen therapy. To investigate the mechanisms behind this paradox, we determined the differences between the transcriptional profiles of ER-α36 and full-length ER-α (ER-α66) in breast cancers and matched normal tissues. Methods: We analyzed ER-α36 and ER-α66 messenger RNA ( mRNA) levels in 74 pairs of breast cancers and matched normal tissues using a real-time quantitative polymerase chain reaction (PCR) assay, and correlated the results with their clinicopathological characteristics. Results: Breast cancers expressed lower ER-α36 mRNA levels than matched normal tissues regardless of their ER-α66 expression status. Down-regulation of ER-α36 mRNA was correlated with local progression, lymph node metastasis, and advanced cancer stage. The level of ER-α66 mRNA was lower in ER-α negative breast cancers compared with matched normal tissues. No differences in ER-α66 mRNA levels were observed during cancer progression. Conclusion: Down-regulation of ER-α36 is associated with carcinogenesis and progression of breast cancer.

ERβ1基因及其剪切变异体真核表达载体的构建与鉴定

目的应用基因克隆技术构建雌激素受体β1(ERβ1)及其5-8外显子缺失变异体ERβ△5-8(文中简称ERβ△),为ERβ功能的研究奠定基础。方法组织中提取总RNA,RT-PCR法扩增基因ERβ1及ERβ△,限制性酶切并克隆于以带EGFP绿色荧光基因及Myc报告基因的非融合真核细胞表达质粒IRES2-EGFP,菌群转化扩增后琼脂糖凝胶电泳,限制性酶切后行DNA测序鉴定。结果 PCR扩增及限制性酶切法证实重组真核表达质粒的构建成功。测序表明ERβ1长度为1593 bp,ERβ△长度为972 bp,与GeneBank上公布的蛋白编码区序列一致。结论成功构建ERβ1基因及其剪切变异体ERβ△真核表达载体。