Determining tamoxifen sensitivity using primary breast cancer tissue in collagen-based three-dimensional culture

We developed a three-dimensional assay prepared from primary breast cancer tissue and quantified tumor response to tamoxifen therapy. Freshly harvested breast cancer biopsies obtained at the time of curative surgical resection were fragmented and embedded into collagen I cushions. Changes in proliferation, apoptosis and tumor volume in response to tamoxifen treatment were quantified using image analysis software and optical projection tomography. Individual and collective invasion of epithelial cells into the surrounding collagen I was observed over the course of the experiment using phase contrast light microscopy and histopathological methods. Addition of tamoxifen to preparations derived from ER+ tumors demonstrated a range of response as measured by proliferative and apoptotic markers. In keeping with published data, tamoxifen reduced the percentage of apoptotic cells expressing cleaved caspase-3 (p = 0.02, Poisson regression analysis). Tamoxifen also reduced residual epithelial volume in ER+ tumors (p = 0.001, Mann-Whitney test), but not in ER low/- tumors (p = 0.78). Changes in tumor volume, as measured by optical projection tomography, allowed stratification into responsive and non-responsive tumors. The model mirrors observations of breast cancer response and histopathological changes to tamoxifen in neo-adjuvant trials. This assay provides a method of screening a battery of therapeutics against individual cancers, informing subsequent design of neo-adjuvant trials.     

The matrix of human breast tumor cells is mitogenic for fibroblasts.

The basis of the scirrhous reaction to human breast carcinoma was investigated. When normal human skin fibroblasts were plated on the preformed extracellular matrix of human breast tumor cells, a remarkable series of changes was observed. The matrix of the tumor cells was mitogenic for the fibroblasts. An increased growth rate and a fourfold increase in cell density was observed. There was also a change in cell morphology and in the pattern in which the cells grew, with an apparent loss of contact inhibition. The spindle-shaped fibroblasts became more elliptical and grew in a series of whorls and dense ridges with spaces between them. These observations were made with the use of newborn foreskin fibroblasts and the matrix of an established line of human breast cancer cells, ZR75-1. No such effect was seen when fibroblasts were plated on their own preformed matrix, on the matrixes of other cell types, on various type-specific collagen gels, or on a combination of collagen and fibronectin or when fibroblasts were grown in media conditioned by the ZR75-1 cells. A floating tumor cell matrix added to the cell media also did not provide the mitogenic stimulus. Apparently, fibroblasts required direct contact with the tumor cell matrix for the mitogenic response to occur. In vivo, the matrix of breast tumor cells may modulate the growth and the morphology of host stromal cells. Collagen is a major synthetic product of fibroblasts. The stimulation of stromal cells to proliferate by adjacent breast tumor matrix may be the basis of the desmoplastic reaction, the intense fibrotic response associated with human breast cancer.     

Appendicular metastatic involvement in breast cancer

The paper describes a case of appendicular metastatic involvement in breast cancer. The 60-year-old female patient developed the signs of acute appendicitis 18 years after detection of primary tumor; after appendectomy the intraoperative appendular specimens exhibited trains of tumor cells that infiltrated the mid- and lower muscle layer third. Immunohistochemical study revealed a moderate expression of Er and Pgr receptors (150 H scores), a positive reaction with EMA, K7, K6, and lactoalbumin The authors review the data available in the literature on the specific features of metastatic spread into the appendix and on management tactics in these patients.     

Results of treatment of breast cancer with tamoxifen]

There were 62 cases of mammary carcinoma treated with the antioestrogen tamoxifen. The age of the patients ranged from 35 to 75 years. Tamoxifen was administered in a daily dosage of 30 mg (3 x 1 tablet). Treatment was carried out as monotherapy or adjuvant therapy. Treatment was carried out for 6 months with a follow-up period of further 6 months. In the group of patients with mammary carcinoma, 23 were premenopausal without metastases after radical mastectomy and sterilisation. The remaining 39 in this group were in the menopause with metastases which were in some cases untreated and in some cases treated by mastectomy, radiation and chemotherapy (CMF-scheme). All Patients were PD in relation to further radiation and/or chemotherapy. Karnofsky-index was 60 minimum. After 4 weeks' treatment with tamoxifen both an improvement in general wellbeing and a regression of the focus of the tumour was achieved. A significant improvement in wellbeing and tumour status could be established for a total of 48 cases of mammary carcinoma. Treatment was well tolerated. Only 1 case of side effects in the form of vomiting occurred.     

Effects of tamoxifen on the cytology of the uterine cervix in breast cancer patients

Tamoxifen, a nonsteroidal antiestrogen, is the endocrine therapy of choice for all stages of breast cancer. Because tamoxifen is well tolerated and has minimal side effects, it is currently being evaluated in large scale trials as a chemopreventive agent for women at risk for developing breast cancer. The potential adverse effects of tamoxifen, specifically the development of proliferative lesions of the endometrium, coupled with the prospect of its wider use, places new emphasis on recognizing tamoxifen-associated histologic and cytologic changes in the female genital tract. The current study evaluated cervical smears from 52 breast cancer patients treated with tamoxifen compared with 21 smears from breast cancer patients who had not received tamoxifen. Cytologic diagnoses were classified according to the Bethesda system. The presence of blood, inflammation, and hormonal effect were also assessed. No squamous intraepithelial lesions were identified. A total of 21 of 38 smears (55%) from patients receiving tamoxifen alone and 11 of 14 smears (78%) from women who received tamoxifen in combination with adjuvant cytotoxic chemotherapy showed atypias compared with only 6 of the 21 breast cancer patients (28%) who did not have hormonal therapy. The number of smears showing atypia was equally divided into changes interpreted as benign reactive and atypical squamous cells of undetermined significance (ASCUS). Of the 19 patients whose smears were classified as ASCUS, 13 patients had a subsequent cervical biopsy and none showed dysplasia or diagnostic human papilloma virus changes. Tamoxifen therapy was not associated with an increase in the presence of blood or inflammation, and no discernible alteration in the hormonal state was seen in the cervical smears. We conclude that the use of tamoxifen may be associated with benign squamous atypia in cervical smears and that the atypia is not associated with intraepithelial lesions. Diagn. Cytopathol. 1998;19:417–422. © 1998 Wiley-Liss, Inc.     

Ionotropic glutamate receptors regulating labeled acetylcholine release from rat striatal tissue in vitro: possible involvement of receptor modulation in magnesium sensitivity

This study evaluated the role of glutamate ionotropic receptors on the control of [3H]acetylcholine ([3H]ACh) release by the intrinsic striatal cholinergic cells. [3H]-choline previously taken up by chopped striatal tissue and converted to [3H]ACh, was released under stimulation by glutamate, N-methyl-d-aspartate (NMDA), kainate and a-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA). Experiments were conducted in the absence of choline uptake inhibitors or acetylcholinesterase inhibitors. A paradigm of two stimulations was employed, the first in control conditions and the second after 9 min of perfusion with the test agents MK-801, 2-amino-5-phosphonopentanoic acid (AP-5), tetrodotoxin (TTX), 6,7-dinitroquinoxaline-2,3-dione (DNQX), 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo-[f]quinoxaline-7-sulfonamide (NBQX), glycine and magnesium. Our results support that (1) in the absence of Mg2+, NMDA is the most effective agonist to stimulate [3H]ACh release from striatal slices (2) magnesium effectively antagonized kainate and AMPA stimulation suggesting that at least part of the kainate and AMPA effects might be attributed to glutamate release (3) besides NMDA, kainate receptors showed a more direct involvement in [3H]ACh release control based on the smaller dependence on Mg2+ and less inhibition by TTX and (4) stimulation of ionotropic glutamate receptors may induce long lasting biochemical changes in receptor/ion channel function since the effects of TTX and/or Mg2+ ions on [3H]ACh release were modified by previous exposure of the tissue to agonists.     

Autophagy and nuclear changes in FM3A breast tumor cells after epirubicin, medroxyprogesterone and tamoxifen treatment in vitro.

OBJECTIVE: Autophagy is a form of physiological programmed cell death which is observable after hormonal withdrawal. In this study, the FM3A murine breast tumor cell line was treated with epirubicin alone and with medroxyprogesterone acetate (MPA) or tamoxifen, to determine if structural and kinetic signs of autophagy may also occur in an enhanced manner during epirubicin sensitization via hormonal agents. METHODS: One-week soft agar colony growth, 96-hour values of plating and pulse thymidine labeling and electron microscopical examinations were performed following treatment with MPA and tamoxifen alone or with epirubicin. RESULTS: Tamoxifen induced signs of autophagy, which was enhanced when it was combined with MPA. Epirubicin also induced autophagy of secretory granules, which coalesced to form an intracytoplasmic lumen. Combining MPA with epirubicin enhanced the formation of apoptotic blebs and chromatin fragmentation. When epirubicin was combined with tamoxifen, peculiar nuclear structures were formed. CONCLUSIONS: Hormonal agents may modulate anthracycline activity towards specific patterns in eliciting cell death, via autophagy and/or as yet unknown nucleolus-specific interactions.     

肌纤维母细胞及在肿瘤间质中激活的纤维母细胞

背景：肌纤维母细胞从1971年被发现以来一直是一个充满争议的课题,近年的研究认识到,该细胞在肿瘤的发生、发展过程中可能起着重要作用。 目的：阐述肌纤维母细胞的生物学特征,以及肿瘤间质纤维母细胞在肿瘤的生长、侵袭方面的作用。 方法：由第一作者检索万方数据库及Pubmed数据库中1990-01/2010-01与肌纤维母细胞相关的文献,在标题和摘要中以“肌纤维母细胞;肿瘤;转化生长因子;侵袭性”或“myofibroblast;tumor;TGF-β;aggressiveness”为检索词进行检索。选择内容与肌纤维母细胞及肿瘤间质纤维母细胞相关的文章,同一领域文献则选择近期发表或发表在权威杂志文章。初检得到116篇文献,根据纳入标准选择41篇文章进行综述。 结果与结论：肌纤维母细胞具有平滑肌和纤维母细胞的特征,肿瘤间质肌纤维母细胞是癌细胞衍生的细胞因子调控成纤维细胞转化而来,通过直接刺激癌细胞的增殖和生存来促进肿瘤生长,并通过水解细胞基质和刺激肿瘤细胞运动来发生侵袭。肌纤维母细胞的出现有助于肿瘤的早期诊断及探索肿瘤治疗的新途径。     

Possible involvement of the endocannabinoid system in memory modulation effect of general anesthetics

Amnesia is a fundamental component of a proper general anesthetic. The mechanism of anesthetic-induced amnesia remains poorly understood. Nowadays, intraoperative awareness and postoperative cognitive dysfunction are two distressing problems receiving increased attention by clinicians, patients and the general public. Extensive evidence indicates that general anesthetics cause amnesia by working on hippocampus and basolateral amygdala (BLA). Recently, evidence from studies in experimental animals has shown that either intra-hippocampus or intra-BLA injection of endogenous cannabinoid receptor 1 (CB1) drugs result in significant changes of cognitive function. In addition, several general anesthetics (i.e. propofol, etomidate and isoflurane) have been reported to interact with the endocannabinoid system. However, there are few studies about whether the CB1 receptor system is involved in anesthetic-induced amnesia. We hypothesize that the CB1 receptor activity in hippocampus and BLA might be regulated by general anesthetics. Once the involvement of the endocannabinoid system in anesthetic-induced amnesia is proved, it will provide a new way to prevent and treat post-traumatic stress disorder caused by intraoperative awareness and postoperative cognitive dysfunction in the future.