The expression of neurokinin-1 and preprotachykinin-1 in breast cancer cells depends on the relative degree of invasive and metastatic potential

Breast cancer has a predilection for metastasis to the bone marrow. The preprotachykinin-I (PPT-I) gene has a central role in the early migration of breast cancer cells into the bone marrow, making this organ a latent repository of the cancer cells. This study investigated whether the invasive and metastatic potential of breast cancer cells correlate with the expression of the PPT-I gene and the receptors for its peptides, neurokinin-1 (NK-1) and NK-2. The studies compared cells that are non-tumorigenic (MCF12A), low metastatic and invasive potential (MCF7), and sublines of MCF with increased invasive and metastatic potential (LCC1 and LCC2). LCC2, but not LCC1 is tamoxifen resistant. Quantitative RT-PCR showed increased expression of PPT-I, NK-1 and NK-2 mRNA LCC1 and LCC2. MCF7 required stimulation by phorbol ester for NK-1 induction. The levels of NK-2 mRNA were significantly increased in LCC2. Clonogenic assays with specific receptor antagonists showed a predominant role for NK-2 in the proliferation of both LCC1 and LCC2. While the growth rate of LCC1 and LCC2 were similar, the latter showed increased migration. Use of a nude mouse model confirmed higher metastatic potential of LCC2, including increased migration to regions of the endosteum. Overall, these studies show a correlation between three neuroendocrine-related genes: PPT-I, NK-1 and NK-2 and the metastatic potential of specific breast cancer cells. These cells provide a model for future studies on bone marrow metastasis.This work was done at UMDNJ-New Jersey Medical School, Departments of Pathology and Laboratory Medicine and Medicine-Hematology/Oncology.     

神经降压素及其受体1与乳腺癌

神经降压素是一种分布于脑和胃肠道的多肽,具有广泛的中枢和外周效应。近来研究表明神经降压素及其1型受体的信号与乳腺癌的增殖、抗凋亡、侵袭转移方面具有密切的联系;而神经降压素受体1仅在乳腺癌中表达将为开发神经降压素类似物为基础的新型肿瘤显像剂及靶向治疗药物提供广泛的应用前景,有助于乳腺癌的早期诊断和治疗。     

P物质/神经激肽1受体系统在肿瘤治疗中的研究进展

P物质（SP）是第1个被发现的速激肽家族成员,广泛分布于哺乳动物的中枢神经系统和外周神经系统,参与多种生理和病理过程。神经激肽受体（NKRs）包括NK1R、NK2R和NK3R,它们是G-蛋白耦联受体家族成员。SP是NK1R内源性的高亲和性和高选择性配体。越来越多的实验结果表明,SP/NK1R系统参与了肿瘤的发生过程。因此,NK1R有可能成为肿瘤治疗的新靶点。我们就SP/NK1R系统在肿瘤中的表达水平,以及SP/NK1R系统作为新靶点在肿瘤治疗领域的研究进展做一简要综述。     

急性坏死性胰腺炎肠粘膜损害与NK-IR的过度表达相关

目的:研究神经激肽-1受体(NK-1R)和神经激肽-2受体(NK-2R)在急性坏死性胰腺炎(ANP)大鼠末端回肠组织中的表达, 探讨该受体的表达与ANP肠粘膜损害的关系。方法:健康成年Sprague-Dawley大鼠随机分为假手术对照组(50只)和ANP组(80只)。假手术对照组开腹后只翻动胰腺, ANP组大鼠胰胆管恒速逆行注射5%牛磺胆酸钠, 制成ANP大鼠模型。应用逆转录聚合酶链反应(RT-PCR)检测末端回肠组织NK-1R和NK-2R的mRNA水平, 应用Westernblot检测NK-1R的蛋白表达水平。结果:与假手术对照组相比, ANP末端回肠组织中NK-1R和NK-2R的mRNA水平过度表达。NK-1R的表达水平分别与肠粘膜的病理学评分(r=0.77, P＜0.01)和肠粘膜通透性(r=0.68, P＜0.01)呈明显正相关。结论:ANP时, 肠组织中NK-1R和NK-2R的表达水平明显上调, P物质的过度作用加剧ANP时肠粘膜的病理损害, 损害肠粘膜屏障的功能。     

Oestrogen receptor and its potential role in breast cancer development

The first studies of oestrogen receptor (ER) focused on measurements of levels in breast cancers and relationship to hormonal response, but the introduction of antibodies which can be used on fixed tissue has meant that ER can be studied in detail in normal and precancerous tissue. Cloning and sequencing of ER has resulted in more detailed understanding of structure and function, with the identification of variant forms in both tumours and normal, followed by the discovery that ER has two forms, the classical ER-α and ER-β. Analyses of both forms in normal and preneoplastic breast will be important for our understanding of the role of ER in the development of breast cancer and its possible prevention. Copyright © 1999 John Wiley & Sons, Ltd.     

Prognostic role of Amphiregulin and the correlation with androgen receptor in invasive breast cancer

BackgroundIn androgen-sensitive prostate cancer, androgenic stimulation induces the synthesis of amphiregulin (AREG). Research is lacking on the role of AREG in invasive breast cancer and the co-expression with androgen receptor (AR) status.Materials and methodsThe present study investigated the prognostic role of AREG in invasive breast cancer cases (N = 298) and the co-expression with the AR status as analysed by immunohistochemistry (IHC).ResultsThe samples were divided into groups according to AREG expression levels: low/no expression (AREG^low/no) and high expression (AREG^high). As shown by cytoplasmic immunostaining, 46.0% (137/298) of invasive breast cancers were AREG^high, and 54.0% (161/298) of cases were AREG^low/no. Co-expression of the AR and AREG accounted for 62.4% (186/298) of cases. A Kaplan–Meier analysis revealed that AREG^high and AR⁺/AREG^high decreased patients’ overall survival (OS) (P = 0.002 and P = 0.006, respectively) and disease-free survival (DFS) (P < 0.001 and P < 0.001, respectively). In Cox models, AR⁺/AREG^high remained an independent prognostic indicator of OS and DFS in invasive breast cancer (hazard ratio [HR], 0.591, 95% confidence interval [CI], 0.407-0.859, P = 0.006; HR, 0.449, 95% CI, 0.236-0.853, P = 0.014, respectively). AREG^high remained an independent prognostic indicator of OS and DFS in estrogen receptor (ER)-negative tumours (P < 0.05).ConclusionsThis study suggested that AREG and the AR were co-expressed in invasive breast cancer. Thus, AREG and the AR may be valuable prognostic biomarkers in invasive breast cancer and promising therapeutic targets, especially in ER-negative breast cancer.     

Down-regulation of BRMS1 by DNA hypermethylation and its association with metastatic progression in triple-negative breast cancer

Breast cancer metastasis suppressor 1 (BRMS1) is a metastasis suppressor gene in several solid tumors. However, the expression and function of BRMS1 in triple-negative breast cancer (TNBC) have not been reported. In this study, we found that BRMS1 was down-regulation in breast cancer cell lines and primary TNBC, while decreased expression of BRMS1 mRNA was significantly associated with lymph node metastasis. And this down-regulation was found to be in accordance with aberrant methylation of the gene. Hypermethylation of the gene was observed in 53.4% (62/116) of the TNBC primary breast carcinomas, while it was found in only 24.1% (28/116) of the corresponding nonmalignant tissues. In addition, BRMS1 expression was restored in MDA-MB-231 after treatment with the demethylating agent, 5-aza-2-deoxycytidine (5-Aza-dC), and demethylation of the highly metastatic cells MDA-MB-231 induced invasion suppression of the cells. Furthermore, the suppression of BRMS1 by siRNA transfection enhanced cancer cells invasion. Collectively, our results suggest that the aberrant methylation of BRMS1 frequently occurs in the down-regulation of BRMS1 in TNBC and that it may play a role in the metastasis of breast cancer.     

大鼠中央杏仁核内微量注射P物质抑制对大鼠胃电-机械活动

目的观察大鼠中央杏仁核内微量注射P物质（SP）对胃肌电和胃运动幅度的影响。方法用铂金丝双电极和应变片传感器引导胃肌电及胃运动信号,用MacLab系统记录并处理信号。结果中央杏仁核内注射4g／L的SP 1μL,可明显抑制胃电．机械活动;注射1．5g／L的SP受体拮抗剂（DPDPDT）1μL,可阻断内源性SP的作用,胃电-机械活动增强;并且DPDPDT可使外源性SP对胃电-机械活动的抑制效应减弱;用0．5g／L阿托品1μL阻断M受体后,SP对胃电-机械活动的抑制效应也减弱。结论SP在中央杏仁核可能通过与SP受体结合,抑制胃电-机械活动,而胆碱能神经元可能参与了SP抑制胃电-机械活动这一过程。     

Amplified in breast cancer 1 expression in breast cancer

Aims: The amplified in breast cancer 1 (AIB1), steroid receptor co‐activator family member, acts as an oestrogen receptor (ER) co‐activator. Acting with HER‐2, it is thought to play a role in endocrine resistance by facilitating ER–growth factor crosstalk. The aim was to analyse AIB1 expression by immunohistochemistry and study its correlations with other prognostic variables in breast cancer and its effect on survival. Methods: A tissue microarray comprising tumours from 438 patients with 15.4 years’ median follow‐up was used. Interpretable AIB1 expression obtained in 395 patients was analysed along with other prognostic factors in breast cancer. Results: AIB1 expression scores ranged from 0 to 30; positive AIB1 expression (score > 14) was seen in 146/395 breast cancers; it correlated negatively with ER (P = 0.003) and progesterone receptor (PR) (P = 0.007), and positively with HER‐2 (P = 0.005) and tumour grade (P = 0.014). It did not correlate with nodal status (P = 0.437). Among ER+ patients, AIB1 expression showed a trend towards loss of PR expression (29% versus 20%; P = 0.14). AIB1 did not predict survival on univariate or multivariate analysis. Conclusions: AIB1 expression correlates with HER‐2 expression in breast cancer and shows a trend of association with loss of PR expression in ER+ tumours. Our study supports the postulated role of AIB1 in ER–growth factor interactions.     

Immunohistochemical localization of substance P and substance P receptor (NK1) in the olivary pretectal nucleus of the rat

The olivary pretectal nucleus (OPN) is the first central nucleus in the pupillary light reflex arc (PLR). Substance P (SP) is a neuropeptide present in the OPN. The present immunohistochemical study, performed at the ultrastructural level, aimed to determine the synaptic localization of SP and SP receptor in the OPN. Three types of SP-positive terminals were found. The most abundant type was of retinal origin, characterized by electron-lucent mitochondria and round vesicles, organized in glomerular structures, making asymmetric synaptic contacts with dendrites, and profiles containing pleomorphic vesicles, also making synaptic contacts with dendrites. The second type of SP-immunoreactive terminal contained electron-dense mitochondria and pleomorphic vesicles. This type made symmetric synaptic contacts and may originate from the ventral part of the lateral geniculate nucleus. The third type of SP-immunoreactive terminals contained electron-dense mitochondria, clear round vesicles, and made an asymmetric synaptic contact. This type originates from the contralateral OPN. SP receptors of the NK1 subtype were revealed to be on dendrites and were part of the glomerular-like arrangement. On account of the present observations, it can be concluded that retinal projections to the OPN use SP as a neuromodulator and synapse on NK1 receptor-containing dendrites of large neurons projecting to the Edinger-Westphal nucleus. Since SP also modulates the parasympathetic component of the PLR, we postulate that SP plays a modulating role in all components of the PLR.     

Low PBRM1 identifies tumor progression and poor prognosis in breast cancer

Background: To investigate the expression and role of PBRM1 in breast cancer, and to evaluate the clinical and prognostic significance of PBRM1 protein in patients with breast cancer. Methods: The expression of PBRM1 was examined in breast cancer tissue and paired non-cancerous tissues by real-time PCR. Moreover, PBRM1 protein expression was evaluated by immunohistochemistry in 150 paraffin-embedded breast cancer specimens. The correlation between PBRM1 expression and clinicopathological features were statistically analyzed. Results: The status of PBRM1 protein in breast cancer tissues is much lower than that in paracarcinoma tissues. Low PBRM1 expression was positively correlated with tumor stage (P =0.003) and lymph node metastasis (P =0.013). The overall (P =0.003) and recurrent-free survival (P =0.001) of the patients with high PBRM1 expression was significantly lower than the low PBRM1 expression group. Multivariate analysis showed that the expression of PBRM1 was an independent factor of overall survival for the patients with breast cancer (P =0.030). Conclusions: PBRM1 might involve in the development and progression of breast cancer as a tumor suppressor, and thereby may be a valuable prognostic marker for breast cancer patients.     

大鼠脊髓白质后索内P物质受体阳性神经元的形态特征及其联系

本研究应用免疫组织化学方法和免疫荧光组织化学双标技术,观察了大鼠脊髓白质后索内的P物质(SP)受体(SPR)阳性神经元的形态特征及其联系。结果表明,脊髓白质后索内存在SPR阳性神经元,它们的胞体较小,常集中在两侧后索的中线上,呈三角形、圆形和多极形;它们的短树突在胞体周围呈放射状,但向后索表面行走的树突较直,且可达脊髓表面;在激光共聚焦显微镜下可见这些SPR阳性神经元呈NeuN阳性,但GFAP呈阴性;它们的胞体及其突起与SP、谷氨酸脱羧酶(GAD)、脑啡肽(ENK)和5-HT阳性纤维及终末形成紧密接触。上述结果说明脊髓白质后索内存在神经元,且呈SPR阳性;这些SPR阳性神经元的活动可能受到多种来源神经信号的调控。     

人参皂甙-Rd对SNI大鼠痛敏异常及脊髓背角内P物质和NK-1受体表达的影响

目的:观察人参皂甙-Rd(Ginsenoside-Rd,G-Rd)对坐骨神经分支选择损伤(spared nerve injury,SNI)大鼠痛敏异常及脊髓背角内P物质(substance P,SP)和NK-1受体表达的影响,进而探讨G-Rd镇痛的脊髓机制.方法:成年雄性SD大鼠(30只)随机分成五组:空白对照组(bl...     

维生素D受体基因多态性与乳腺癌的相关性

目的：研究维生素D受体基因的多态性与乳腺癌的关系。方法：收集86例乳腺癌患者及134名对照,用聚合酶链反应－限制性片段长度多态性方法,在维生素D受体基因的3’端分析了两个限制性酶切位点（ApaI及TaqI）的多态分布。结果：发现TaqI位点等位基因在两个群体间分布的差异有显著性（P＝0．0004）。进一步对基因型进行分析发现,Tt、tt基因型与乳腺癌相关。而ApaI位点两等位基因未发现在两群体中存在差异。对ApaI及TaqI两座位的单体型进行分析,发现tA间存在连锁不平衡。在两群体中分析单体型的分布发现tA在病例中的比例明显高于对照人群,提示tA单体型与乳腺癌相关。两个位点等位基因及单体型与临床指标的分析均未发现阳性结果。结论：维生素D受体基因的多态怀与乳腺癌有关,提示维生素D受体基因与乳腺癌有关。     

Expression of insulin-like growth factor 1 receptor in primary breast cancer: immunohistochemical analysis

Insulin-like growth factor-1 receptor (IGF-1R) has been implicated in regulation in tumor growth. The results of previous studies performed by radioimmunoassay are conflicting, and the prognostic significance of IGF-1R expression in primary breast cancer is still controversial. IGF-1R expression was evaluated in formalin-fixed, paraffin-embedded tissue of 210 primary breast cancer patients by using anti-IGF-1R antibody. The clinicopathologic variables and 5-year disease-free survival were studied, and their correlations between IGF-1R expressions were investigated. IGF-1R overexpression was observed in 43.8% of tumors. IGF-1R overexpression had no correlation with prognosis or with other clinicopathologic parameters, such as age, tumor size, nodal status, histologic grade, hormone receptor status, and human epidermal growth factor 2 status. Though its prognostic value in breast cancer is limited, immunohistochemical evaluation of IGF-1R by using this monoclonal antibody may be useful in translational research using archived material.     

Androgen receptor expression in breast cancer patients tested for BRCA1 and BRCA2 mutations

Pristauz G, Petru E, Stacher E, Geigl J B, Schwarzbraun T, Tsybrovskyy O, Winter R & Moinfar F
(2010) Histopathology 57, 877–884 Androgen receptor expression in breast cancer patients tested for BRCA1 and BRCA2 mutations Aim: To assess the expression of receptors for androgen (AR), oestrogen (ER) and progesterone (PR) as well as human epidermal growth factor receptor type 2 (Her‐2/neu) status of breast carcinomas in breast cancer susceptibility gene (BRCA) BRCA1/2 mutation carriers and BRCA1/2 negative tested women. Methods: One hundred and thirty‐five breast cancers in women tested for BRCA1/2 mutations. Screening for BRCA1 and BRCA2 mutations was performed by direct sequencing of all BRCA1 and BRCA2 exons as well as the surrounding intronic sequences. Additionally, BRCA genes were analysed with multiplex ligation‐dependent probe amplification. Consecutive paraffin sections were examined immunhistochemically for AR, ER, PR and Her‐2/neu. Results: Of the 135 tumours, 43 (32%) were BRCA1‐related, 18 (13%) were BRCA2‐related and 74 (55%) were BRCA1/2‐negative. Seventy‐two per cent of the BRCA1‐related, 22% of the BRCA2‐related and 12% of the BRCA1/2‐negative tumours were triple (ER, PR, Her2neu)‐negative. Eighty‐four per cent of BRCA1 mutated cancers were high‐grade (G3) tumours. ARs were expressed in 30% (13 of 43) of BRCA1‐related, in 78% (14 of 18) in BRCA2‐related tumours and in 76% (56 of 74) in BRCA1/2 negative tumours. Twenty‐one per cent of ER‐negative BRCA1‐related tumours expressed androgen receptors. Conclusion: Approximately one in five BRCA1 mutated breast cancers negative for ER and PR express androgen receptors. Modulation of AR might open a new avenue for treating these high‐risk cancers.     

Assessment of hormone receptor status in breast cancer

The aim of the present paper was to investigate the most adequate method for the assessment of hormone receptor status in breast cancer in routine clinical settings. Subjects were 486 patients with primary breast cancer who underwent surgery and postoperative tamoxifen monotherapy in 1982-1993. Using representative sections of the primary lesion in each patient, estrogen receptors (ER) were immunohistochemically stained. Patients were divided into ER-positive and ER-negative groups using various methods, and then overall and 5 year recurrence-free survival rates were compared. The results of ER status, which are diagnosed on entire cancer area and invasive cancer area, matched in 98% of cases. When assessing prognosis based on the proportion of positive cells, a significant difference in 5 year recurrence-free survival was seen between ER-positive and ER-negative patients for a cut-off of 10%, and in overall and 5 year survival for a cut-off of 33%. Based on the proportion and the intensity of positive cells (Allred score), a significant difference was seen in overall and 5 year survival for a cut-off in total scores between 4 and 5 points. When assessing hormone receptors of breast cancer in routine clinical settings, it is sufficient to determine the proportion of positive cells in the entire cancer area.