Estrogen, estrogen receptors, and hepatocellular carcinoma: Are we there yet?

A protective role of the sex steroid hormone estrogenin hepatocellular carcinoma(HCC) was suggested a few decades ago according to clinical data showing higher HCC morbidity and mortality among males. Several recent studies further confirmed the anti-cancer effects of estrogen in the liver. However, it remains to be identified how to exploit estrogen signalling within clinical settings for HCC treatment. There are several unresolved issues related to the estrogen pathway in liver cells. The main problems include the absence of a clear understanding of which estrogen receptor(ER) isoform is predominantly expressed in normal and malignant liver cells, the ER isoform expression difference between males and females, and which ER isoform should be targeted when designing HCC therapy. Some of those questions were recently addressed by Iyer and coauthors. The current editorial review critically analyses the study by Iyer et al(WJG, 2017) that investigated the expression of ER subtypes in liver samples collected from patients with a healthy liver, hepatitis C virus cirrhosis, and HCC. ER presence was evaluated in association with gender, intracellular localization, inflammation marker NF-kB, and proliferation-related effector cyclin D1. The study limitations and advantages are discussed in light of recent advances in the HCC and estrogen signalling areas.

Estrogen receptor expression in chronic hepatitis C and hepatocellular carcinoma pathogenesis

AIM To investigate gender-specific liver estrogen receptor(ER) expression in normal subjects and patients with hepatitis C virus(HCV)-related cirrhosis and hepatocellular carcinoma(HCC).METHODS Liver tissues from normal donors and patients diagnosed with HCV-related cirrhosis and HCV-related HCC were obtained from the NIH Liver Tissue and Cell Distribution System. The expression of ER subtypes, ERα and ERβ, were evaluated by Western blotting and real-time RT-PCR. The subcellular distribution of ERα and ERβ was further determined in nuclear and cytoplasmic tissue lysates along with the expression ofinflammatory [activated NF-κB and IκB-kinase(IKK)] and oncogenic(cyclin D1) markers by Western blotting and immunohistochemistry. The expression of ERα and ERβ was correlated with the expression of activated NF-κB, activated IKK and cyclin D1 by Spearman's correlation. RESULTS Both ER subtypes were expressed in normal livers but male livers showed significantly higher expression of ERα than females(P < 0.05). We observed significantly higher m RNA expression of ERα in HCV-related HCC liver tissues as compared to normals(P < 0.05) and ERβ in livers of HCV-related cirrhosis and HCV-related HCC subjects(P < 0.05). At the protein level, there was a significantly higher expression of nuclear ERα in livers of HCV-related HCC patients and nuclear ERβ in HCV-related cirrhosis patients as compared to normals(P < 0.05). Furthermore, we observed a significantly higher expression of phosphorylated NF-κB and cyclin D1 in diseased livers(P < 0.05). There was a positive correlation between the expression of nuclear ER subtypes and nuclear cyclin D1 and a negative correlation between cytoplasmic ER subtypes and cytoplasmic phosphorylated IKK in HCV-related HCC livers. These findings suggest that dysregulated expression of ER subtypes following chronic HCVinfection may contribute to the progression of HCVrelated cirrhosis to HCV-related HCC.CONCLUSION Gender differences were observed in ERα expression in normal livers. Alterations in ER subtype expression observed in diseased livers may influence genderrelated disparity in HCV-related pathogenesis.

Role of sex steroid receptors in pathobiology of hepatocellular carcinoma

The striking gender disparity observed in the incidence of hepatocellular carcinoma (HCC) suggests an important role of sex hormones in HCC pathogenesis. Though the studies began as early as in 1980s,the precise role of sex hormones and the significance of their receptors in HCC still remain poorly understood and perhaps contribute to current controversies about the potential use of hormonal therapy in HCC. A comprehensive review of the existing literature revealed several shortcomings associated with the studies on estrogen receptor (ER) and androgen receptor (AR) in normal liver and HCC. These shortcomings include the use of less sensitive receptor ligand binding assays and immunohistochemistry studies for ERα alone until 1996 when ERβ isoform was identified. The animal models of HCC utilized for studies were primarily based on chemical-induced hepatocarcinogenesis with less similarity to virus-induced HCC pathogenesis. However,recent in vitro studies in hepatoma cells provide newer insights for hormonal regulation of key cellular processes including interaction of ER and AR with viral proteins. In light of the above facts,there is an urgent need for a detailed investigation of sex hormones and their receptors in normal liver and HCC. In this review,we systematically present the information currently available on androgens,estrogens and their receptors in normal liver and HCC obtained from in vitro,in vivo experimental models and clinical studies. This information will direct future basic and clinical research to bridge the gap in knowledge to explore the therapeutic potential of hormonal therapy in HCC.

Liver expression of steroid hormones and Apolipoprotein D receptors in hepatocellular carcinoma

AIM: To evaluate the tissular expression of Androgen (A), Estrogen (E) and Progesterone (Pg) receptors, and Apolipoprotein D (ApoD), in liver tumors from resected hepatocellular carcinoma (HCC) cases in order to assess their possible relationship to prognosis. METHODS: We performed an immunohistochemical study using tissue microarrays (containing more than 260 cancer specimens, from 31 HCC patients and controls) to determine the presence of specif ic antibodies against AR, ER, PgR and ApoD, correlating their findings with several clinico-pathological and biological variables. The staining results were categorized using a semi-quantitive score based on their intensity, and the percentage of immunostained cells was measured. RESULTS: A total of 21 liver tumors (67.7%) were positive for AR; 16 (51.6%) for ER; 26 (83.9%) for PgR and 12 (38.7%) stained for ApoD. We have found a wide variability in the immunostaining score values for each protein, with a median (range) of 11.5 (11.5-229.5) for AR; 11.1 (8.5-65) for ER; 14.2 (4-61) for PgR; and 37.7 (13.8-81.1) for ApoD. A history of heavy ethanol consumption, correlated positively with AR and PgR and negatively with ER status. HCV chronic infection also correlated positively with AR and PgR status. However, the presence of ApoD immunostaining did not correlate with any of these variables. Tumors with a positive immuno-staining for PgR showed a better prognosis. CONCLUSION: Our results indicate a moderate clinical value of the steroid receptor status in HCC, emphasizing the need to perform further studies in order to evaluate the possible role of new hormonal-based therapies.

Comprehensive Analysis of Estrogen Receptor 1 Dysregulation in Liver Hepatocellular Carcinoma: Implications for Prognosis and Therapeutic Targeting - A Secondary Publication

The study investigates the expression pattern and regulatory mechanisms of estrogen receptor 1 (ESR1) in liver hepatocellular carcinoma (LIHC) through comprehensive bioinformatics analysis. Utilizing UALCAN and GEPIA2 databases, significant down-regulation of ESR1 expression is observed in LIHC samples compared to normal controls, indicating its potential role in tumor progression. Further analysis reveals consistent down-regulation across different clinical variables including patient age, gender, race, and various stages of LIHC, affirming the regulatory role of ESR1 in tumor development and progression. Additionally, promoter methylation analysis demonstrates hypermethylation of ESR1 in LIHC samples, negatively correlating with its expression. This association persists across different clinical parameters, emphasizing the inverse relationship between ESR1 methylation and expression levels. Survival analysis indicates that up- regulation of ESR1 is associated with better overall survival, suggesting its potential as a prognostic biomarker in LIHC. Furthermore, genetic mutation analysis using cBioPortal reveals a spectrum of alterations in ESR1, including amplification, missense mutation, deep deletion, splice mutation, and truncating mutation, highlighting the genetic complexity of ESR1 in LIHC. These findings collectively contribute to a deeper understanding of ESR1 dysregulation in LIHC and its clinical implications as a potential therapeutic target and prognostic marker.

Therapeutic Value of Estrogen Receptorαin Hepatocellular Carcinoma Based on Molecular Mechanisms

The incidence of hepatocellular carcinoma(HCC)is signifi-cantly lower in women than men,implying that estrogen receptors(ERs)may play an important role in this sex di-morphism.Recently,considerable progress has been made in expanding our understanding of the mechanisms of ERs in HCC.As one of the most important ERs,ERαfunctions as a tumor suppressor in the progression of HCC through various pathways,such as STAT3 signaling pathways,lipid metabolism-related signaling pathways,and non-coding RNAs.However,the function of ERαwas reduced with the changes of some molecules in the liver,which may develop further into HCC and make it difficult to achieve an effective hormone treatment effect.Intriguingly,there are signs that individualized hormone therapy according to the activity of ERαwill overcome this challenge.Based on these observa-tions,it is particularly imperative to reassess and extend the function of ERα.In this review,we mainly elucidated molecular mechanisms associated with ERαin HCC and investigated the individualized hormone therapy based on these mechanisms,with the aim of providing new insights for HCC treatment.

Female hepatology:Favorable role of estrogen in chronic liver disease with hepatitis B virus infection

Chronic hepatitis B virus (HBV) infection is the most common cause of hepatic fibrosis and hepatocellular carcinoma (HCC),mainly as a result of chronic necroinflammatory liver disease. A characteristic feature of chronic hepatitis B infection,alcoholic liver disease and nonalcoholic fatty liver disease (NAFLD) is hepatic steatosis. Hepatic steatosis leads to an increase in lipid peroxidation in hepatocytes,which,in turn,activates hepatic stellate cells (HSCs). HSCs are the primary target cells for inflammatory and oxidative stimuli,and these cells produce extracellular matrix components. Chronic hepatitis B appears to progress more rapidly in males than in females,and NAFLD,cirrhosis and HCC are predominately diseases that tend to occur in men and postmenopausal women. Premenopausal women have lower hepatic iron stores and a decreased production of proinflammatory cytokines. Hepatic steatosis has been observed in aromatase-deficient mice,and has been shown to decrease in animals after estradiol treatment. Estradiol is a potent endogenous antioxidant which suppresses hepatic fibrosis in animal models,and attenuates induction of redox sensitive transcription factors,hepatocyte apoptosis and HSC activation by inhibiting a generation of reactive oxygen species in primary cultures. Variant estrogen receptors are expressed to a greater extent in male patients with chronic liver disease than in females. These lines of evidence suggest that the greater progression of hepatic fibrosis and HCC in men and postmenopausal women may be due,at least in part,to lower production of estradiol and a reduced response to the action of estradiol. A better understanding of the basic mechanisms underlying the sex-associated differences in hepatic fibrogenesis and carciogenesis may open up new avenues for the prevention and treatment of chronic liver disease.

高原鼢鼠睾丸雌激素受体免疫组化染色最适浓度抗体的筛选

【目的】筛选高原鼢鼠(Eospalax baileyi)睾丸雌激素受体(Estrogen receptor,ER)免疫组化染色的适宜抗体浓度。【方法】研究选取10只成年雄性高原鼢鼠的睾丸,采用免疫组织化学SP法(IHC-SP法)检测雌激素受体α(ERα)和雌激素受体β(ERβ)在1∶150、1∶200和1∶250稀释浓度下的染色效果。【结果】ERα抗体稀释浓度1∶250的免疫组化平均光密度值显著高于1∶150和1∶200的抗体稀释浓度免疫组化平均光密度值(P<0.05),而ERβ抗体稀释浓度1∶200的平均光密度值最高(P<0.01),最适浓度结果均表现出阳性细胞染色强度高、背景清晰。【结论】高原鼢鼠睾丸中ERα和ERβ抗体稀释浓度分别为1∶250与1∶200的免疫组化染色效果最佳。

大脑皮质及海马雌、孕激素受体在癫痫发作后的变化

目的 探讨雌、孕激素受体与癫痫发病的关系。方法 采用免疫组织化学 ABC法观察了侧脑室注射谷氨酸致痫雄性大鼠相关脑区雌、孕激素受体的变化。结果 雌、孕激素受体广泛分布于大脑皮质及海马区域 ,定位于胞核及胞浆 ,致痫组大鼠雌激素受体较正常对照组明显降低 ,而孕激素受体较正常对照组明显升高。结论 雌。

原发性肝细胞癌中雌激素受体α与β的表达及临床意义

目的 :研究雌激素受体 ( ER) α与 β在原发性肝细胞癌中的表达水平及分布规律。方法 :用免疫组织化学方法检测了 63例原发性肝细胞癌及 63例非肝癌肝组织的 ERα与 ERβ的表达。结果 :原发性肝细胞癌中 ERα与 ERβ的阳性率分别为 2 7% ( Edmondson分级 级的肝癌为 2 3%、 级为 2 8%、 级为 2 9% )和 1 3% ( Edmondson分级 级的肝癌为31 %、 级为 8%、 级为 7% ) ,非肝癌肝组织中 ERα与 ERβ的阳性率分别为 32 % (肝硬化 :30 %、肝海绵状血管瘤 :32 %、正常肝组织 :5 0 % )和 2 9% (肝硬化 :30 %、肝海绵状血管瘤 :2 6%、正常肝组织 :2 5 % )。肝癌与非癌肝组织比较 ERα阳性率无显著差异 ( P>0 .0 5 ) ,但ERβ阳性率有显著意义 ( P<0 .0 5 ) ;Edmondson分级 级的肝癌 ERβ的阳性率与 、 级肝癌比较 ,统计学差异有显著性 ( P<0 .0 5 ) ;肝癌组内 ERα阳性较 ERβ有显著差异 ( P<0 .0 5 )。结论 :原发性肝细胞癌可能具有一定的激素依赖性 ,肝癌的发生、发展及预后可能与ERα的表达无关 ,但与肝癌组织中

肝癌组织中雌激素受体基因启动子甲基化及临床研究

目的:研究原发性肝细胞癌(hepa-tocellular carcinoma,HCC)组织中ER基因启动子甲基化修饰与临床病理特征间的关系,同时观察5-杂氮胞苷对ERmRNA表达的影响。方法:分别采用MSP和RT-PCR方法检测30例HCC及癌旁组织中ER基因启动子甲基化修饰状况及ERmRNA的表达状况,并用5-杂氮胞苷处理肝癌细胞系SMMC-7721和HepG2。结果:30例HCC组织中ER启动子甲基化修饰阳性率为60·0%(18/30),而30例癌旁组织中ER基因启动子甲基化修饰阳性率为30·0%(9/30),两者差异有统计学意义,χ2=5·46,P=0·037。30例HCC组织中ERmRNA阳性表达12例(40·0%),其中甲基化组织和未甲基化组织中ERmRNA表达率分别为22·2%和66·7%。ER启动子甲基化修饰与mRNA表达呈负性相关,χ2=5·93,P=0·024。同时,ER启动子区甲基化修饰与肝癌患者血清AFP含量增高呈正相关,χ2=12·13,P=0·001。细胞培养结果提示低浓度的5-杂氮胞苷可诱导肝癌细胞系ERmRNA重新表达。结论:肝癌组织中ER启动子甲基化修饰是个频繁事件,因此沉默ER基因的表达有可能促进肝癌的发展。

肝癌组织雌激素受体表达与生物学特征的研究

目的：探讨雌激素受体（ｅｓｔｒｏｇｅｎ ｒｅｃｅｐｔｏｒ， ＥＲ）在肝细胞肝癌（ｈｅｐａｔｏｃｅｌｌｕｌａｒ ｃａｒｃｉｎｏｍａ， ＨＣＣ）中的表达及其与ＨＣＣ生物学特征的关系。方法：用免疫组织化学方法检测了４３例ＨＣＣ及对应癌旁肝组织及４例正常肝组织的雌激素受体的表达，经大体及显微镜检查了这些肝细胞肝癌的病理特征。结果：雌激素受体阳性率在肝细胞肝癌为 ３７．２１％，癌旁肝组织为６０．４６％，正常肝组织 １００％；雌激素受体阳性肝癌６２．５％为单结节，５６． ２５％有完整包膜，８１．２５％无血管侵犯，其构成比高于雌激素阴性肝癌的 ２５.９２％、２２．２２％和 ４８．１５％，统计学差异有显著性；小肝癌的雌激素受体阳性率为６６.６７％显著高于大肝癌的２５．８１% （Ｐ＜０．０５）。结论：肝细胞肝癌可能具有一定的激素依赖性，且雌激素受体阳性肝癌与雌激素阴性肝癌相比，前者的生物学行为和预后较好。

雄激素受体、雌激素受体在肝细胞型肝癌中的免疫电镜观察

目的：观察雄激素受体（ＡＲ）与雌激素受体（ＥＲ）在肝癌和非癌肝组织超微结构上的分布。探讨ＡＲ、ＥＲ在肝癌致癌过程中的作用。方法：用免疫胶体金染色法（ＩＧＳ）标记１３例肝细胞型肝癌与５例非癌肝组织细胞内雄激素受体（ＡＲ）及雌激素受体（ＥＲ），并用免疫透射电镜观察。结果：发现胶体金颗粒主要分布于线粒体双层膜和脊膜；粗面内质网；结合及游离核蛋白体；核膜及核周间隙，异染色质及常染色质等部位。统计分析表明，肝癌细胞金颗粒明显多于非癌肝细胞内金颗粒（Ｐ＜０．０５）。肝癌细胞胞质、胞核ＡＲ金颗粒明显多于ＥＲ金颗粒（Ｐ＜０．０５）。而非癌肝细胞胞质、胞核ＡＲ、ＥＲ ｋ颗粒数量无显著差别（Ｐ＞０．０５）。结论：提示肝癌是一种雄激素依赖肿瘤，为抗性激素治疗肝癌提供了理论依据。

肝癌细胞雌激素受体显性阴性突变体ERδ5基因的分离及生物学功能分析

临床流行病学研究显示,ERα基因第5外显子缺失突变体ERδ5在原发性肝细胞癌特异性高表达,它被认为是判断肝癌预后的最重要的新型分子标志.本研究从肝癌细胞分离雌激素受体突变体ERδ5基因,并对其进行克隆、表达,探讨其分子生物学特性及生物学功能.通过反转录PCR,从肝细胞分离出ERδ5基因并将其克隆到真核表达载体,通过体外翻译及Western印迹验证该基因成功表达;通过细胞荧光技术检测了ERδ5在肝细胞中的定位;利用荧光素酶报告基因检测技术确证ERδ5对ERα转录活性的影响.结果发现,从肝癌细胞分离出长1113bp的ERδ5基因,该基因编码蛋白在体外不稳定,易于降解.在肝癌细胞中,ERδ5蛋白主要定位在细胞核,与ERα定位极为相似,但在生物学功能上,ERδ5能够明显干扰ERα的转录活性.研究结果表明,ERδ5作为显性阴性突变体,在肝癌细胞中直接抑制ERα的转录活性.

脂肪细胞雌激素受体的表达及其与人体脂肪分布的相关性

目的:研究脂肪细胞雌激素受体在人体脂肪细胞中的表达情况及其与人体脂肪分布的关系。方法:应用免疫组织化学法检测64例成人腹部皮下脂肪和腹腔内大网膜脂肪细胞中雌激素受体蛋白的定量表达情况。结果:成人脂肪组织中雌激素受体蛋白表达阳性;在女性腹腔内大网膜脂肪细胞中雌激素受体表达明显高于男性,在皮下脂肪中二者无明显差异。结论:人体脂肪细胞核内有雌激素受体蛋白的表达,脂肪细胞可能是雌激素作用的靶器官;人类脂肪分布的性别差异可能与雌激素和雌激素受体作用相关。

原癌基因C-met和雌激素受体在子宫内膜癌中的表达及相关性的探讨

目的 了解子宫内膜癌组织中原癌基因 C- m et和雌激素受体 (ER)的表达状况 ,探讨内膜癌中 C- met的表达与 ER的关系。方法 采用免疫组化 SP法。结果判断采用组织化学记分 (H值 ) ,H值≥ 70为阳性表达。结果 宫内膜癌中 C- met表达随手术 -病理分期及组织学分级的增加而增强 ,ER的表达随着手术 -病理分期及组织学分级的增加而减弱。C- met表达与 ER呈负相关趋势 (r=- 0 .5 84 2 ,P<0 .0 5 )。结论 在子宫内膜癌中 ,C- m et的表达与 ER表达呈负相关 ,两者联合使用对宫内膜癌的激素治疗 。

HBV阳性与HBV阴性的肝癌细胞株雌激素受体α与β的表达

目的探讨雌激素受体(ER)α与β在HBV阳性和HBV阴性的肝癌细胞株中的表达情况。方法用免疫组织化学方法检测HBV阳性的肝癌细胞株QGY-7701、QGY-7703、Hep-G2.2215和HBV阴性的肝癌细胞株Hep-G2、SMMC-7721、HHCC中ERα与ERβ的表达。结果HBV阳性的肝癌细胞株ERα与ERβ的表达显著低于其在HBV阴性的细胞株中的表达(P<0.05)。结论ERα与ERβ可能在HBV诱发肝癌的过程中起一定作用。

他莫昔芬对人肝癌细胞增殖及ER、Smad4表达的影响

目的:研究他莫昔芬对人肝癌HepG2细胞增殖、雌激素受体(ER)、Smad4表达的影响。方法:根据他莫昔芬的不同浓度分为3组,即7.5、15、30μmol/L组,对照组不加他莫昔芬,每组设6个平行对照孔,每孔加入细胞浓度为1×105个/ml的细胞悬液0.1 ml,培养24、48、72 h。采用MTT法测定OD值,然后计算抑制率;免疫组化染色观察他莫昔芬对人肝癌HepG2细胞ER、Smad4表达的影响。结果:他莫昔芬抑制人肝癌细胞增殖,随浓度的增加及处理时间的延长,抑制率增加;他莫昔芬抑制肝癌细胞ER表达,促进Smad4的表达,具有时间-浓度依赖。结论:他莫昔芬可通过影响肝癌细胞ER、Smad4的表达来抑制肝癌细胞增殖。

芳香化酶P450、ERβ在宫颈癌组织的表达及临床意义

目的探讨芳香化酶P450、ERβ在正常宫颈组织、宫颈癌组织及癌周组织的表达差异,进而探讨其临床意义。方法对西安交通大学医学院第二附属医院2004年至2008年手术切除的正常宫颈组织、宫颈癌组织及癌周组织存档蜡块标本各30例,用免疫组化SP法检测其表达。结果芳香化酶P450在正常宫颈组织、癌周组织和宫颈癌组织的表达率分别为0%(0/30)、0%(0/30)和36.7%(11/30),经比较有显著性差异(P<0.05),而正常宫颈组与癌周组织组的表达无显著性差异(P>0.05)。ERβ在正常宫颈组织、癌周组织和宫颈癌组织的表达率分别为73%(22/30)、70%(21/30)和36.7%(11/30),相比较有显著性差异(P<0.05),而正常宫颈组织与癌周组织的表达无显著性差异(P>0.05)。结论芳香化酶P450在宫颈癌组织的阳性表达可能参与宫颈癌的发生;ERβ在宫颈癌组织的阳性表达率低,可作为预测宫颈癌变及宫颈癌预后良好的一项参考指标。

人肝细胞性肝癌性激素受体的表达

目的 研究人肝细胞性肝癌组织、癌旁组织和正常肝组织中雄激素受体 (AR)和雌激素受体 (ER)的表达水平及分布规律。方法 应用单克隆抗体免疫组化法检测 5 0例肝细胞性肝癌标本的肿瘤组织、癌旁组织和 10例肝内胆管结石的肝组织中 AR和 ER的表达情况。结果 在肝癌组织、癌旁组织和肝内胆管结石的肝组织中 ,AR的阳性表达率分别为 30 %、8%和 0 % ,除癌组织显著高于癌旁组织外 (P<0 .0 1) ,其它各组无显著性差异 (P>0 .0 5 ) ;ER的阳性表达率分别为 12 %、10 %和2 0 % ,在三组中均无显著性差异 (P>0 .0 5 )。在所有阳性片中 ,AR和 ER的标记指数 (L I)均 <2 5。结论 AR和