乳腺癌原发肿瘤和复发肿瘤间ER、PR和HER-2表达不一致及其对患者生存的影响

目的探讨乳腺癌原发肿瘤和复发肿瘤组织中ER、PR和HER-2表达不一致与临床病理特征的关系及对预后的影响。方法采用免疫组化和荧光原位杂交法检测62例乳腺癌原发肿瘤和复发肿瘤组织中ER、PR和HER-2的表达,分析乳腺癌原发肿瘤和复发肿瘤组织中ER、PR和HER-2表达不一致与临床病理特征的关系及与患者生存的关系。结果在62例乳腺癌原发肿瘤和复发肿瘤组织中ER、PR和HER-2表达不一致分别为6例(9. 7%)、9例(14. 5%)和3例(4. 8%)。ER表达不一致患者的中位总生存期和中位复发后生存期分别为29个月和6个月,而ER表达一致患者的中位总生存期和中位复发后生存期分别为44个月和15个月(P=0. 021,P=0. 027)。PR及HER-2表达不一致患者与PR及HER-2表达一致患者总生存期和复发后生存期差异均无统计学意义(P均 0. 05)。结论乳腺癌原发肿瘤和复发肿瘤组织之间ER、PR和HER-2表达状态显著改变。乳腺癌ER状态不稳定提示患者预后较差。     

一种新型转移诱导蛋白AGR2在乳腺癌中的表达及其临床和预后意义

目的 研究转移诱导蛋白AGR2在人乳腺癌中表达和意义及其与患者预后的关系.方法 采用组织芯片,应用免疫组织化学方法检测160例乳腺癌和20例乳腺良性病变组织AGR2表达,以及乳腺癌ERα、PR和HER2表达状况,并用Kaplan-Meier曲线法和Log-rank检验对有随访资料的127例进行与AGR2的相关性分析.结果 乳腺癌AGR2表达量较乳腺良性病变显著性增高(68.3%和25.0%,P＜0.01);AGR2表达与乳腺癌组织学分级呈负相关(P＜0.05),与乳腺癌ERα和PR表达呈正相关(分别为P＜0.05和P＜0.01);Logistic回归模型显示AGR2和TNM分期是ERa阳性乳腺癌淋巴结转移重要的影响因子(均P＜0.01);Kaplan-Meier生存曲线显示ERα阳性乳腺癌患者中,AGR2阳性患者总生存率和无复发生存率均显著低于阴性患者(均P＜0.01),COX比例风险模型分析也证实AGR2蛋白表达是ERα阳性乳腺癌患者独立的预后影响因子(P＜0.01).结论 AGR2表达异常可能参与了乳腺癌的发生、发展过程,该机制可能部分依赖雌激素受体途径发挥其诱导肿瘤转移作用.AGR2可能是一个潜在的分子标记物,对雌激素受体阳性乳腺癌患者的预后评估有一定提示作用.     

雌激素受体α阴性乳腺癌雌激素受体α基因启动子区CpG岛甲基化和肼苯哒嗪去甲基化作用

目的 检测雌激素受体(ER)α阴性乳腺癌细胞株MDA-MB-231和MDA-MB-435细胞及ERα阴性乳腺癌组织中ERα基因启动子区CpG岛甲基化状态;探索肼苯哒嗪能否作为去甲基化药物恢复ERα基因表达。方法应用特异性聚合酶链反应(MSP)检测乳腺癌细胞株MDA-MB-231和MDA-MB-435细胞和20例ERα阴性乳腺癌组织ERα基因3个启动子区A、B、CpG岛甲基化情况,肼苯哒嗪处理上述两种乳腺癌细胞,逆转录(RT)-PCR检测不同启动子调控下ERα基因异型体(isoform)ERα-A、ERα-B、ERα-C mRNA和ERα基因公共编码区mRNA表达。结果MDA-MB-231和MDA-MB-435细胞启动子区ERα-A、ERα-B均存在CpG岛甲基化,ERα-C无甲基化,20例ERα阴性乳腺癌组织中,13例(65％)ERα-A、10例(50％)ERα-B CpG岛甲基化阳性。其中9例ERα-A、ERα-BCpG岛甲基化均阳性(45％),仅1例(5％)ERα-C存在CpG岛甲基化。肼苯哒嗪处理上述两种细胞后,检测到ERα-A、ERα-B mRNA和公共编码区mRNA表达。结论乳腺癌组织和细胞ERα基因表达沉默可能与ERα基因启动子区A、B甲基化有关,且肿瘤分期愈晚,甲基化程度愈高。肼苯哒嗪能作为去甲基化药物诱导ERα基因表达。     

乳腺癌并发肥胖症患者血浆瘦素水平与乳腺癌组织中瘦素mRNA的表达及临床病理的关系

目的:检测乳腺癌并发肥胖症患者乳腺癌组织中瘦素(leptin)mRNA的表达情况和血浆leptin水平,并探讨两者之间及其与临床病理之间的关系。方法:对124例居住在武汉检查者同时采集病史、进行体格检查并留取血浆,测定其leptin水平。采用半定量逆转录-聚合酶链反应(RT-PCR)检测瘦素mRNA的表达。结果:乳腺癌并发肥胖组血浆leptin水平(12.02μg.L-1±1.23μg.L-1)显著高于乳腺良性病变并发肥胖组(9.84μg.L-1±0.98μg.L-1)及单纯肥胖组(9.79μg.L-1±1.16μg.L-1)(P<0.05)。瘦素mRNA在乳腺癌并发肥胖组中的表达水平(0.71±0.32)明显高于乳腺良性病变并发肥胖组(0.41±0.26)(P<0.05)。血浆lep-tin水平与乳腺组织中瘦素mRNA表达水平显著正相关(r=0.4220,P=0.0180)。血浆leptin和瘦素mRNA的表达与腋窝淋巴结转移、绝经、TNM分期及病理类型均无显著相关性。结论:leptin可能参与了乳腺癌的发生。     

视黄酸受体RARs在不同乳腺组织中的表达模式

目的观察视黄酸受体RARs在正常、良性和恶性乳腺组织中的表达模式。方法运用半定量PCR技术,对视黄酸受体RARs不同亚型在不同乳腺组织中的表达情况进行检测,检测结果进行统计学分析。结果RARα在不同乳腺组织的表达无明显差异,但在ER阳性的恶性组织中的表达却明显高于ER阴性的恶性组织,结果有统计学意义;RARβ在本实验40例组织中没有检测到表达;RARγ的表达在肿瘤组织中明显增高,有统计学意义,但在良、恶性组织中没有差异。结论RARα与ER的表达相关,可与ER同样作为乳腺癌的一个预后指标;RARγ与细胞的增殖有关,但与分化无明显关系。     

乳腺癌合并肥胖症患者血浆瘦素水平与乳腺癌组织中瘦素受体mRNA的表达及临床病理的关系

检测乳腺癌合并肥胖症患者乳腺癌组织中瘦素受体(leptin receptors)mRNA的表达情况和血浆瘦素水平,并探讨二者之间及其与临床病理之间的关系.对124例单纯肥胖者,或乳腺癌合并肥胖者,或乳腺良性病变合并肥胖者同时采集病史、进行体格检查并留取血浆,测定其瘦素的水平.采用半定量逆转录-聚合酶链反应(RT-PCR)检测瘦素受体mRNA的表达.结果显示,乳腺癌合并肥胖组血浆瘦素水平显著高于乳腺良性病变合并肥胖组及单纯肥胖组(P＜0.05).瘦素长型受体[Lep-R(L)]mRNA及瘦素短型受体[Lep-R(S)]mRNA在乳腺癌合并肥胖组中的表达水平均明显高于乳腺良性病变合并肥胖组(P＜0.05).血浆瘦素水平与乳腺组织中Lep-R(L)mRNA及Lep-R(S)mRNA水平呈显著正相关.血浆瘦素和瘦素受体mRNA的表达与腋窝淋巴结转移、绝经、TNM分期及病理类型均无显著相关性(P＞0.05).结论:瘦素可能参与了乳腺癌的发生.     

乳腺肿瘤组织生长抑素受体检测及其临床意义

探讨乳腺良、恶性肿瘤SSR表达的差异对乳腺肿瘤良、恶性的鉴别诊断的价值, 观察SSR表达与ER、PR表达的相关性, 预测乳腺肿瘤生长抑素受体显像的临床价值. 将手术切下的标本分成4组: 乳腺肿瘤恶性组(恶性组)及其对照组(癌旁组), 乳腺肿瘤良性组(良性组)及其对照组(瘤旁组).采用RBA法测定各组织的SSR表达, 采用免疫LsAB法测定ER、PR表达.结果:(1)恶性组、癌旁组、良性组、瘤旁组SSR表达分别为108.6±67.3fmol/mg pr、37.2±9.6fmol/mg pr、43.4±12.6fmol/mg pr、33.9±10.2fmol/mg pr, 恶性组比其对照组及良性组都高, 差异有显著性(P值分别为＜0.001、＜0.01),而良性组与其对照组则无显著性差异(P＞0.05);(2)SSR与ER、PR表达的相关系数分别为0.859、0.750.因此, 大多数乳腺癌组织表达高密度的SSR,可用SSR显像来做良、恶性肿瘤的初步鉴别, SSR表达与ER、PR表达有良好的相关性, 可预测肿瘤恶性程度及其患者预后.     

子宫肌瘤雌激素受体亚型mRNA的表达

目的　探讨雌激素受体亚型的表达与子宫平滑肌瘤发生、发展的关系。方法　将60例子宫肌瘤患者分为AB两组, A组32例<48岁,B组28例>48岁,应用原位杂交技术测定平滑肌瘤组织中雌激素两种受体亚型mRNA的表达,取肿瘤周 围正常的平滑肌作对照。结果　子宫肌瘤组织中ERαmRNA的表达明显高于正常的平滑肌组织,而ERβmRNA的表达降 低;B组ERαmRNA在子宫平滑肌瘤组织中的表达较A组呈下降趋势,而ERβmRNA的表达呈上升趋势。结论　子宫肌瘤的 发生发展与ERαmRNA的高表达有关,亦与ERβmRNA表达减少有关。肌瘤的萎缩可能与ERβmRNA表达的升高趋势及 ERαmRNA表达的下降趋势均相关。     

miR-145在乳腺癌中表达及意义

目的 研究一种微小RNA(miR-145)在人乳腺癌及乳腺良性病变组织中的表达,分析miR-145的表达与乳腺癌临床病理特征的关系及其对乳腺癌发生发展的意义.方法 采用组织芯片平台,应用原位杂交方法 检测91例乳腺癌和38例乳腺良性病变组织(包括21例纤维腺瘤,17例乳腺腺病)miR-145表达.结果 乳腺癌miR-145高表达率较乳腺良性病变降低(31.9%和60.5%,P<0.01);乳腺癌miR-145高表达率与肿瘤腋淋巴结转移(P<0.01),TNM高分期(P<0.05),c-erbB-2过表达(P<0.01)状况均相关;乳腺癌miR-145表达与患者年龄、肿瘤大小、组织学分级、ER、PR均无相关性(均P>0.05).结论 miR-145表达异常可能参与了乳腺癌的发生、发展过程,是一个潜在的乳腺癌分子标记物.     

乳腺癌及乳腺良性病变中BCL-6 mRNA和蛋白表达及生物学意义

目的研究BCL-6在人乳腺癌及乳腺良性病变组织中的表达情况,探讨BCL-6的表达与乳腺癌患者临床病理特征的关系及其在乳腺癌中的生物学意义。方法采用免疫组化MaxVision两步法检测127例乳腺癌中BCL-6蛋白的表达,另随机选取其中74例采用原位分子杂交(in situ hybridizationI,SH)法检测BCL-6 mRNA的表达。同时分别检测50例乳腺良性病变组织中BCL-6 mRNA和蛋白的表达。结果 BCL-6 mRNA和蛋白在乳腺癌组织中的阳性率分别为54.1%(40/74)和32.3%(41/127),两者呈正相关;50例乳腺良性病变组织中分别有3例和5例的少数腺上皮细胞(<10%)呈BCL-6蛋白和BCL-6mRNA弱阳性。BCL-6蛋白的表达与乳腺癌组织学分级、肿瘤大小及TNM分期均相关(P<0.01),但与患者年龄和腋窝淋巴结转移无关。结论 BCL-6蛋白可能成为乳腺癌中一个新的有价值的分子标记物。     

Evaluation of BrightGen HR RT-qDx assay to detect nuclear receptors mRNA overexpression in FFPE breast cancer tissue samples for selection of tamoxifen therapy

Breast cancer is a significant cause of death in women. Estrogen receptor (ER) and progesterone receptor (PR) are important prognostic factors indicating higher recovery rate in the breast cancer patients. Currently, immunohistochemical (IHC) staining is a conventional method to identify expression of ER and PR. If a breast cancer patient expresses ER or PR, a chemotherapy with estrogen inhibitors such as tamoxifen is supposed to be effective. Although IHC staining is a reliable method, it may not a useful method for continuous monitoring of ER and PR expression changes in multiple breast cancer patients. In the present study, we evaluated an alternative method of IHC for detection of ER and PR expression. A quantitative RT-PCR method called ‘the BrightGen HR RT-qDx assay’ was employed to detect mRNA expression of the nuclear receptors in 199 formalin-fixed paraffin-embedded (FFPE) breast cancer tissue samples. Among the ER/PR positive samples by IHC, 83 were determined positive and 16 were determined negative for the nuclear receptor mRNA by the quantitative RT-PCR method. Among the ER/PR negative samples by IHC, 37 were determined negative and 2 were determined positive by the quantitative RT-PCR method. The overall sensitivity and specificity of the quantitative RT-PCR method were 83.8% and 94.8% (P = 0.0026), respectively. We also optimized the quantitative RT-PCR method by setting up the diagnostic cut-off value using the likelihood ratio. The highest likelihood ratio was when the expression levels of the relative nuclear receptor mRNA passed 103.3 at which sensitivity and specificity was highest. These data suggest that BrightGen HR RT-qDx assay could be an alternative method for detection of the prognostic factors of nuclear receptors expressed in breast cancer patients for providing essential information for therapeutic application of tamoxifen.     

Oestrogen regulated gene expression in normal and malignant endometrial tissue

OBJECTIVES: The aim of this study was to examine the expression of oestrogen regulated genes in premenopausal and postmenopausal normal and malignant endometrial specimens. The molecular mechanisms and the role of these genes in endometrial carcinogenesis are poorly understood. METHODS: Normal and malignant endometrial specimens were collected from patients undergoing hysterectomy. Real time TaqMan PCR was used to examine the mRNA expression levels of oestrogen receptor a (ERa) and b (ERb), progesterone receptor (PR), insulin like growth factor 1 (IGF-1) and vascular endothelial growth factor (VEGF). RESULTS: Expression analysis was carried out on 60 patients. ERa was more predominantly expressed in the endometrial samples than ERb, 28% of the specimens did not express ER. Normal pre and postmenopausal tissue expressed higher levels of ERa, PR and IGF-1 than malignant tissue. ERa and PR expression was significantly higher in the proliferative phase endometrium compared to the secretory phase (P < 0.05). PR mRNA expression was significantly correlated with ERa in all tissue types. CONCLUSIONS: ERa expression may play an important role in the regulation of PR in normal and malignant endometrium. Further work is needed to establish if IGF-1 plays a role in a subset of endometrial cancers and if isoforms of VEGF play a role in endometrial cancer.     

KDM5B基因在乳腺癌中的表达及其临床意义

目的:研究KDM5B基因在乳腺癌组织中的表达情况,并探讨其表达差异与患者临床病理资料和预后的关系。方法:从肿瘤基因组图谱(TCGA)数据库中收集113例正常乳腺组织和1 090例乳腺癌数据集,下载KDM5B mRNA表达谱资料;收集中山大学附属第三医院甲乳外科2015年～2016年乳腺癌切除标本共90例,采用real-time PCR的方法检测乳腺癌及其癌旁组织中KDM5B mRNA的表达量,取中位数分为高表达组与低表达组,分析其与临床资料及病例特征的关系;利用Kaplan-Meier plotter分析KDM5B mRNA表达与乳腺癌患者预后的相关性。结果:KDM5B在乳腺癌中的表达均显著高于正常乳腺组织(P 0. 01)。在TCGA的乳腺癌数据中,KDM5B的表达与人表皮生长因子受体2(HER2)、雌激素受体(ER)、年龄、病理组织类型及淋巴结转移显著相关(P 0. 01),与孕激素受体(PR)、绝经及远处转移无显著相关。在我们收集的乳腺癌样本中,KDM5B的表达与HER2、年龄及淋巴结转移显著相关(P 0. 05),而与ER、PR、绝经、病理组织类型和远处转移无显著相关。KDM5B表达越高,乳腺癌患者总生存时间(HR=1. 39,P=0. 005)和无病生存时间(HR=1. 32,P 0. 001)越短。结论:在乳腺癌组织中KDM5B高表达,且与患者的预后相关; KDM5B的表达与乳腺癌患者HER2表达、年龄和淋巴结转移显著相关。     

乳腺癌耐药相关蛋白在乳腺癌的表达及与临床病理参数相关性

目的探讨原发性乳腺癌中乳腺癌耐药相关蛋白(BCRP)表达与临床指标及病理参数的相关性。方法采用免疫组织化学方法检测BCRP及雌激素受体α(ERα)、孕激素受体(PR)、HER-2、P53等病理参数的表达,应用Kruskal-Wallis秩和检验和Spearman相关性检验分析BCRP与临床指标及病理参数的相关性。结果原发性乳腺癌中BCRP表达阳性率64.39%,癌旁组织中BCRP表达阳性率33.33%,两者相比有显著性差异(X~2=9.323,P=0.002);BCRP表达水平与患者年龄、病理类型、临床分级及淋巴结转移无关,而与患者是否绝经有关,未绝经患者乳腺癌组织中BCRP表达水平显著高于已绝经患者(X~2=6.928,P=0.008);BCRP表达水平与ERα(r=0.204,P=0.019)和HER-2表达水平呈正相关(r=0.246,P=0.004),与PR、P53表达水平不具有相关性。结论乳腺癌中BCRP蛋白表达水平明显高于癌旁正常组织,其表达强度与与患者体内雌激素水平及ERα及HER-2强度正相关。     

乳腺癌中跨膜蛋白81基因高表达不利于患者预后

目的　揭示乳腺癌组织中跨膜蛋白81（TMEM81）的表达变化及其与预后的关系。 方法　使用TCGA数据库的门户网站cBioPortal、UALCAN以及癌症多组学和临床数据库LinkedOmics分析TMEM81在乳腺癌组织中的变化及其与临床预后的关系。 结果　cBioPortal分析结果显示26%乳腺癌患者癌组织中TMEM81基因发生了改变,包括突变、拷贝数畸变和mRNA水平上调, mRNA水平上调超过默认阈值（EXP≥2）的比例为18%,且TMEM81 mRNA水平上调不利于患者预后（P<0.05）;UALCAN分析结果显示乳腺癌组织中TMEM81 mRNA水平显著高于正常组织（P<0.01）;LinkedOmics分析结果显示癌组织TMEM81 mRNA水平分别受到乳腺癌PAM50分型、雌激素受体、孕激素受体、病理分期、病理T分期、组织学类型、种族和年龄的影响（P<0.01, P<0.01,P<0.01,P<0.01,P<0.01,P<0.01,P<0.01,P<0.05）。最后, TMEM81 mRNA水平与其基因拷贝数畸变正相关（P<0.01）,而与其DNA甲基化水平负相关（P<0.01）。 结论　乳腺癌组织中TMEM81基因拷贝数增加和甲基化水平降低促进其表达,TMEM81高表达不利于预后,可作为乳腺癌预后的候选标志物。     

ACLY: A biomarker of recurrence in breast cancer

ObjectiveACLY is a cytoplasmic metabolic enzyme involved in lipid synthesis. It also affects proliferation and metastasis of breast cancer. However, the correlation of ACLY expression with breast cancer recurrence is unclear.MethodsThe Oncomine and TCGA databases were used to investigate the mRNA expression of ACLY in breast cancer. Immunohistochemistry (IHC) was used to evaluate ACLY expression level in tumor tissues and normal tissues from 127 breast cancer patients. Next, the prognostic role of ACLY was explored by analyzing the clinicopathological features and prognosis during follow-up. The role of ACLY in breast cancer cells drug resistance was further detected by CCK-8 assays and quantitative real-time polymerase chain reaction (qRT-PCR).ResultsACLY mRNA and protein expression was significantly increased in the breast cancer tissues compared to normal tissues. Clinically, high ACLY levels were associated with ER status, PR status, tumor size, TNM stage, and lymph node invasion. Upregulated ACLY predicted worse tumor relapse-free survival (RFS) of breast cancer patients in univariate analyses and in multivariate models. In subgroup analysis, patients with high ACLY expression showed worse RFS in the TNM III or ER positive subgroups. Moreover, ACLY over-expression induced the resistance of breast cancer cells to docetaxel and promoted the expression of multi-drug resistant protein ABCB1/ABCG2.ConclusionsOur study highlights the possibility of ACLY as a potential and independent biomarker for the recurrence prediction in breast cancer patients. It may be related to ACLY promoting drug resistance in breast cancer cells.     

Estrogen receptor β– an independent prognostic marker in estrogen receptor α and progesterone receptor‐positive breast cancer?

Both subtypes of estrogen receptor (ER), ERα and ERβ, are normally present in the mammary gland. The role of ERα as a prognostic marker in breast cancer is well established due to the beneficial effect of providing tamoxifen as adjuvant therapy. The role of ERβ, however, is less clear. To gain insight into the importance of ERβ in breast cancer, 145 primary breast cancers were examined by immunohistochemistry for ERβ, and the expression level was compared with ERα and progesterone receptor (PR) status. Especially, we wanted to examine the significance of ERβ in the contrasting ERα+/PR+ and ERα?/PR? subgroups. In the ERα+/PR+ subgroup (dual positive), the survival difference between patients with low, medium and high ER β level was statistically significant (p = 0.004), with more than 70% of patients with medium and high ERβ levels surviving 100 months, compared with less than 30% in the group with low ERβ level. Further, for ERα+/PR+ patients there was a reduced risk of fatal outcome by multivariate analysis with increasing ERβ levels (p(trend) < 0.01 [univariate analysis]; p(trend) = 0.05 [multivariate analysis]). The risk was 31% and 27% for medium and high ERβ levels, respectively, compared with low ERβ level, adjusting for standard prognostic factors such as tumor diameter, nuclear tumor grade (quantified by mean nuclear area), lymph node status, and patient age at operation. For patients with ERα?/PR? tumors (dual negative), however, there was no association between ERβ levels and patient outcome. Our findings indicate that ERβ expression provides independent prognostic information for breast cancers with ERα/PR‐positive status, a feature typical among screen‐detected breast cancers. The role of ERβ needs to be further evaluated especially in this group of breast cancers.     

Histological and immunohistochemical study of estrogen and progesterone receptors in normal human breast tissue in adult age groups vulnerable to malignancy

Analysis of receptor status has become standard procedure for assessing breast cancer patients. Estrogen causes epithelial proliferation in breast tissue via the estrogen receptor (ER). The progesterone receptor (PR) is also regulated by the estrogen gene. Analyzing ER and PR together gives information regarding the likely response of carcinoma patients to hormonal therapy. The aim of the present study was to record the expression patterns of ER and PR in normal mammary tissue in different age groups to provide reference data to facilitate histological diagnosis. Breast tissues from the upper outer quadrant of each side of 27 adult female cadavers were examined after H & E staining. ER and PR were identified and examined by immunohistochemistry. The percentage area occupied by parenchyma relative to stromal tissue was calculated in different age groups and was about 4:6, 3.5:6.5, 3:7, 2:8, and 1.5:8.5 in the 3rd, 4th and 5th, 6th, 7th, 8th and 9th, and 10th decades of life, respectively. Both ER and PR were present in all age groups and the numbers of both receptors were maximal during the 4th decade. The distribution and staining patterns for both ER and PR were recorded in different age groups. The contiguous pattern of ER, which is considered pathognomonic of breast carcinoma, was not seen except in one case in the 6th decade. Moderately stained ER and PR receptor sites predominated throughout. The study of normal breast tissue of similar age might provide comparisons that will help histopathologists to make clinical diagnoses from breast biopsies. Clin. Anat. 29:729–737, 2016. © 2016 Wiley Periodicals, Inc.