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.

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.

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.

大鼠实验性隐睾复位固定术后雌激素受体α表达的变化

目的：研究实验性大鼠隐睾复位固定术对生精细胞凋亡及雌激素受体α（ERn）表达的影响。方法：建立大鼠左侧隐睾模型，模型建立7d后实施左侧隐睾复位固定术，采用TUNEL法检测睾丸生精细胞凋亡，免疫组织化学和RT-PCR检测各组睾丸ERα表达的变化。结果：隐睾内生精细胞凋亡增加，ERa表达降低；复位固定术能降低生精细胞凋亡，增强睾丸内ERα表达。复位固定术后24d生精细胞凋亡指数明显低于复位固定术前的隐睾组；复位固定术后48d生精上皮组织结构已基本恢复，其凋亡指数及ERα表达与假手术组比较，差异无统计学意义。结论：ERα与隐睾内生精细胞凋亡密切相关，隐睾复位固定术能通过恢复睾丸内ERα的表达，抑制生精细胞凋亡。

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。

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.

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.

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.

Transcriptional activation of Nlp by estrogen-ERa in breast cancer

Estrogen receptor-α(ERα) is the key transcription factor that regulates cell proliferation and homeostasis. In this pathway, estrogen plays an important role in genomic instability and cell cycle regulation processes and the mechanisms of its action are multifaceted. In this study, we showed that estrogen regulates genomic instability through promoting the expression of Nlp, a BRCA1-associated centrosomal protein which is involved in microtubule nucleation, spindle formation, chromosomal missegregation and abnormal cytokinesis. We demonstrated that the expression of Nlp is strongly associated with ERα and FOXA1 level in clinical breast cancer samples with poor clinical outcomes to breast cancer patients.Addition of estrogen in the ER-positive breast cancer cells resulted in elevation of NLP mRNA.Significantly, we identified that estrogen-ERα is capable of regulating Nlp expression through specifically binding ERα to the proximal region and the Estrogen Responsive Elements(ERE) enhancer in the distal region of NLP gene. Reporter assays demonstrated that estrogen directly activated Nlp promoter. ChIP assay results showed that E2-ERα directly bound to the EREs of Nlp. Therefore, overexpression of Nlp in breast cancer exhibits a hormone-dependent pattern, and estrogen participates in the regulation of genome instability and cell cycle in breast cancer cells partially through transcriptional activation of NLP gene. Overexpression of Nlp enhances the malignant progression of ERα-positive breast cancer cells in vitro, whereas knockdown of Nlp suppresses this biological effects in ERα-positive breast cancer cells.ERα/Nlp axis may serve as a promising target against breast cancer.

Environmental microcystin exposure triggers the poor prognosis of prostate cancer: Evidence from case-control, animal, and in vitro studies

Microcystin-leucine-arginine(MC-LR)is positively linked with multiple cancers in humans.However,the association between MC-LR and the risk and prognosis of prostate cancer has not been conducted in epidemiological studies.No reported studies have linked MC-LR exposure to the poor prognosis of prostate cancer by conducting experimental studies.The content of MC-LR was detected in most of the aquatic food in wet markets and supermarkets in Nanjing and posed a health risk for consumers.MC-LR levels in both prostate cancer tissues and serum were significantly higher than controls.The adjusted odds ratio(OR)for prostate cancer risk by serum MC-LR was 1.75(95%CI:1.21-2.52)in the whole subjects,and a positive correlation between MC-LR and advanced tumor stage was observed.Survival curve analysis indicated patients with higher MC-LR levels in tissues exhibited poorer overall survival.Human,animal,and cell studies confirmed that MC-LR exposure increases the expression of estrogen receptor-α(ERα)and promotes epithelial-mesenchymal transition(EMT)in prostate cancer.Moreover,MC-LR-induced decreased E-cadherin levels,increased vimentin levels,and increased migratory and invasive capacities of prostate cancer cells were markedly suppressed upon ERαknockdown.MC-LR-induced xenograft tumor growth and lungmetastasis in BALB/c nude mice can be effectively alleviated with ERαknockdown.Our data demonstrated that MC-LR upregulated vimentin and downregulated E-cadherin through activating ERα,promoting migration and invasion of prostate cancer cells.Our findings highlight the role of MC-LR in prostate cancer,providing new perspectives to understand MC-LR-induced prostatic toxicity.