epidermal-growth-factor and Carcinoma--Intraductal--Noninfiltrating

Ductal carcinoma in situ (DCIS) is an early stage noninvasive breast cancer that originates in the epithelial lining of the milk ducts, but it can evolve into comedo DCIS and ultimately, into the most common type of breast cancer, invasive ductal carcinoma. Understanding the progression and how to effectively intervene in it presents a major scientific challenge. The extracellular matrix (ECM) surrounding a duct contains several types of cells and several types of growth factors that are known to individually affect tumor growth, but at present the complex biochemical and mechanical interactions of these stromal cells and growth factors with tumor cells is poorly understood. Here we develop a mathematical model that incorporates the cross-talk between stromal and tumor cells, which can predict how perturbations of the local biochemical and mechanical state influence tumor evolution. We focus on the EGF and TGF-β signaling pathways and show how up- or down-regulation of components in these pathways affects cell growth and proliferation. We then study a hybrid model for the interaction of cells with the tumor microenvironment (TME), in which epithelial cells (ECs) are modeled individually while the ECM is treated as a continuum, and show how these interactions affect the early development of tumors. Finally, we incorporate breakdown of the epithelium into the model and predict the early stages of tumor invasion into the stroma. Our results shed light on the interactions between growth factors, mechanical properties of the ECM, and feedback signaling loops between stromal and tumor cells, and suggest how epigenetic changes in transformed cells affect tumor progression.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Epidermal Growth Factor; Female; Humans; Mathematical Concepts; Models, Biological; Neoplasm Invasiveness; Signal Transduction; Stromal Cells; Transforming Growth Factor beta; Tumor Microenvironment

A large fraction of ductal carcinoma in situ (DCIS), a non-invasive precursor lesion of invasive breast cancer, overexpresses the HER2/neu oncogene. The ducts of DCIS are abnormally filled with cells that evade apoptosis, but the underlying mechanisms remain incompletely understood. We overexpressed HER2 in mammary epithelial cells and observed growth factor-independent proliferation. When grown in extracellular matrix as three-dimensional spheroids, control cells developed a hollow lumen, but HER2-overexpressing cells populated the lumen by evading apoptosis. We demonstrate that HER2 overexpression in this cellular model of DCIS drives transcriptional upregulation of multiple components of the Notch survival pathway. Importantly, luminal filling required upregulation of a signaling pathway comprising Notch3, its cleaved intracellular domain and the transcriptional regulator HES1, resulting in elevated levels of c-MYC and cyclin D1. In line with HER2-Notch3 collaboration, drugs intercepting either arm reverted the DCIS-like phenotype. In addition, we report upregulation of Notch3 in hyperplastic lesions of HER2 transgenic animals, as well as an association between HER2 levels and expression levels of components of the Notch pathway in tumor specimens of breast cancer patients. Therefore, it is conceivable that the integration of the Notch and HER2 signaling pathways contributes to the pathophysiology of DCIS.

Topics: Animals; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Cell Line; Cell Proliferation; Epidermal Growth Factor; Epithelial Cells; Female; Gene Expression Profiling; Green Fluorescent Proteins; HEK293 Cells; Humans; Immunoblotting; Mammary Glands, Human; Mice; Mice, Transgenic; Microscopy, Confocal; Models, Biological; Oligonucleotide Array Sequence Analysis; Receptor, ErbB-2; Receptor, Notch3; Receptors, Notch; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Signal Transduction; Transfection

Lapatinib, a selective orally available inhibitor of epidermal growth factor receptor (EGFR) and ErbB2 receptor tyrosine kinases, is a promising agent for the treatment of breast cancer. We examined the effect of lapatinib on the development of mammary tumors in MMTV-erbB2 transgenic mice, which express wild-type ErbB2 under the control of the mouse mammary tumor virus promoter and spontaneously develop estrogen receptor (ER)-negative and ErbB2-positive mammary tumors by 14 months of age. Mice were treated from age 3 months to age 15 months with vehicle (n = 17) or lapatinib (30 or 75 mg/kg body weight; n = 16 mice per group) by oral gavage twice daily (6 d/wk). All statistical tests were two-sided. By 328 days after the start of treatment, all 17 (100%) of the vehicle-treated mice vs five (31%) of the 16 mice treated with high-dose lapatinib developed mammary tumors (P < .001). Among MMTV-erbB2 mice treated for 5 months (n = 20 mice per group), those treated with lapatinib had fewer premalignant lesions and noninvasive cancers in their mammary glands than those treated with vehicle (P = .02). Lapatinib also effectively blocked epidermal growth factor-induced signaling through the EGFR and ErbB2 receptors, suppressed cyclin D1 and epiregulin mRNA expression, and stimulated p27 mRNA expression in human mammary epithelial cells and in mammary epithelial cells from mice treated for 5 months with high-dose lapatinib. Thus, cyclin D1, epiregulin, and p27 may represent useful biomarkers of lapatinib response in patients. These data suggest that lapatinib is a promising agent for the prevention of ER-negative breast cancer.

Topics: Animals; Antineoplastic Agents; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Cyclin D1; Epidermal Growth Factor; Epiregulin; ErbB Receptors; Female; Gene Expression Regulation, Neoplastic; Humans; Lapatinib; Mammary Neoplasms, Experimental; Mice; Mice, Transgenic; Precancerous Conditions; Protein Kinase Inhibitors; Quinazolines; Receptor, ErbB-2; Receptors, Estrogen; RNA, Messenger; Signal Transduction

By means of a cytokine release assay we have shown that in cell populations derived from primary breast carcinoma, epidermal growth factor (EGF) is secreted by cells with the characteristic morphological and immunophenotypic profile of activated macrophages (positive for CD68, CD16, CD25). EGF secretion was observed in 11 (31%) of 35 primary tumours. Secretion of EGF by normal or malignant epithelial cells was not observed. We found no association between EGF secretion by the primary tumour and recognised clinical indices of prognosis.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Epidermal Growth Factor; Epithelium; Female; Humans; Immunophenotyping; Leukocyte Common Antigens; Macrophage Activation; Macrophages; Middle Aged

The in vivo administration of somatostatin (SS) or its analogues is capable of suppressing the growth of pancreatic cancer in experimental animals. We examined the effects of SS-14 and its analogue RC-160 on the in vitro growth of two human pancreatic cancer cell lines MiaPaCa-2 and Panc-1 stimulated with epidermal growth factor (EGF) or insulin-like growth factor 1 (IGF-1). Neither SS-14 nor RC-160 inhibited the growth of either cell line. In contrast RC-160 did inhibit the EGF-stimulated growth of a rat pancreatic cancer cell line AR42J. Binding studies with 125I-Tyr11 somatostatin revealed the presence of a single class of high affinity binding sites with a Kd of 0.20 +/- 0.05 nM and a Bmax of 2.1 +/- 0.26 pmoles mg-1 protein on AR42J but not displaceable binding was observed on MiaPaCa-2 or Panc-1. We conclude that lack of receptors accounts for the failure of SS-14 and RC-160 to influence the growth of human pancreatic cancer in vitro. These results, taken together with other findings, lead us to question the therapeutic efficacy of somatostatin and its analogues as mono-therapy in the treatment of human pancreatic cancer.

Topics: Animals; Carcinoma, Intraductal, Noninfiltrating; Epidermal Growth Factor; Humans; Insulin-Like Growth Factor I; Pancreatic Neoplasms; Rats; Receptors, Neurotransmitter; Receptors, Somatostatin; Somatostatin; Tumor Cells, Cultured

A series of mouse mammary epithelial cell lines has been established by a protocol that gives highly reproducible results. The mammary epithelial cell lines, designated as FSK lines, were judged to be epithelial based on positive immunostaining for keratin-intermediate filaments, negative immunostaining for vimentin-intermediate filaments, hormonal induction of casein, and the ability to exhibit ductal and alveolar mammary morphogenesis in vivo. The FSK cell lines are dependent on epidermal growth factor and insulin in a low serum (1%) medium. Conditioned medium from spindle cell cultures replaced the requirement for serum and increased the growth of FSK3 and FSK4 4-5 times in collagen gels and 12-14 times in monolayer culture, respectively. Following injection into the mammary fat pad at passages 2-11, the FSK cell lines generated stable transplantable hyperplastic alveolar outgrowth lines. The in vivo outgrowth lines were judged as preneoplastic based on their stable alveolar morphology in vivo and an increased susceptibility for tumorigenesis. The FSK cell lines and their derivative in vivo outgrowth lines provide a new and potentially productive system to examine critical molecular alterations involved in the development of mammary preneoplasias. Furthermore, the reproducibility of the in vitro culture system provides the assurance that stable cell lines of mouse mammary epithelial cells can be generated easily and at will.

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Animals; Carcinoma, Intraductal, Noninfiltrating; Cell Division; Cell Line; Culture Techniques; Epidermal Growth Factor; Epithelial Cells; Female; Fluorescent Antibody Technique; Keratins; Mammary Glands, Animal; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Precancerous Conditions

The present work was performed to clarify the effects of epidermal growth factor (EGF) on the growth of human breast cancer and carcinoma of the esophagus. Human breast cancer MX-1, UM-1, and esophageal cancer ES-4 were transplanted to the subcutaneous tissue of nude mice. Human EGF (2 micrograms) was injected locally into the subcutaneous tissue surrounding the tumor. The tumor growth was followed for 7 days after treatment, and the estimated tumor weight, tumor doubling time, tumor growth curve, tumor growth rate, and results from histologic examination were used to evaluate the effects of EGF on the growth of tumors. We investigated the dose effect on the growth of these tumors using various concentrations of EGF. We also studied the EGF receptor status of these transplanted tumors and its effect on the influence of EGF treatment. A growth-inhibitory effect was noted in these three tumors with 2 micrograms of EGF. EGF inhibited growth of ES-4 tumor in a dose-dependent manner. Treatment with 2 ng of EGF or saline did not inhibit growth. However treatment with 20 ng or 200 ng of EGF inhibited growth in proportion to the concentrations. All tumors were positive by the EGF receptor binding assay. The efficacy of EGF treatment correlated with EGF receptor levels. EGF receptor levels were also influenced by EGF treatment. These results suggest that human EGF showed an anti-tumor effect on MX-1 and UM-1 breast cancer and also on ES-4 esophageal cancer.

Topics: Animals; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Carcinoma, Squamous Cell; Dose-Response Relationship, Drug; Epidermal Growth Factor; ErbB Receptors; Esophageal Neoplasms; Female; Humans; Male; Mice; Mice, Nude; Neoplasm Transplantation; Transplantation, Heterologous

The pathogeneses of hypercalcemia and hypophosphatemia which developed in a patient with metastatic invasive ductal breast carcinoma were studied. The patient had low plasma levels of immunoreactive parathyroid hormone (PTH) and 1,25(OH)2D, increased nephrogenous cyclic adenosine monophosphate (cAMP) excretion and low TmPO4/GFR, suggesting the presence of humoral PTH-like activity. The tumor extract showed activities which would stimulate bone resorption in vitro and cAMP generation in the osteogenic cell line, MC3T3 E1, and in the rat kidney cortex. In addition, the extract stimulated epidermal growth factor (EGF)-independent colony formation of the NRK 49F cells in soft agar, and inhibited the binding of EGF to A431 cells, indicating it to have transforming growth factor (TGF)-alpha activity. The extract contained appreciable amounts of immunoreactive PTH-related protein (PTH-rP) but negligible amounts of immunoreactive PTH. Thus, the PTH-like activity for stimulating cAMP generation in the bone and kidney was attributed to PTH-rP. Chromatographic analyses on reverse phase high performance liquid chromatography (HPLC) separated the PTH-rP activity from that of TGF-alpha and the bone resorbing activity in vitro was found only in the fractions of PTH-rP. It was concluded that this breast cancer produced PTH-rP as well as TGF-alpha, and the former was thought to have a major role to play in the humoral hypercalcemia of malignancy observed in this patient.

Topics: Adult; Animals; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Chromatography, High Pressure Liquid; Cyclic AMP; Epidermal Growth Factor; Female; Glomerular Filtration Rate; Humans; Hypercalcemia; Kidney; Neoplasm Proteins; Parathyroid Hormone-Related Protein; Rats; Transforming Growth Factors; Tumor Cells, Cultured

Transplantation of fetal salivary mesenchyma into adult mammary glands resulted in atypical outgrowths from the mammary duct system. These duct-alveolus nodules (DAN) were distinguishable from hyperplastic alveolar nodules (HAN) that arose from normal mammary duct systems in mice infected with murine mammary tumor virus (MuMTV). DAN displayed a type of ductal branching characteristic of salivary gland rather than of mammary gland, reflecting a tissue-specific perturbation of epithelium-mesenchyma in DAN in milk-transmitted MuMTV-infected C3H/HeN mice and in MuMTV-negative BALB/c mice given 7,12-dimethylbenz[a]anthracene (DMBA) subsequent to transplantation of fetal salivary mesenchyma. Mammary cancers were not increased in milk-transmitted MuMTV-free C3H/HeN and GRS/A mice that received salivary mesenchyma transplants. Salivary mesenchyma accelerated mammary carcinogenesis by increasing the mammary epithelial cell population responsive to MuMTV and DMBA.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Intraductal, Noninfiltrating; Epidermal Growth Factor; Female; Mammary Neoplasms, Experimental; Mammary Tumor Virus, Mouse; Mice; Mice, Inbred Strains; Precancerous Conditions; Salivary Glands; Transplantation, Isogeneic