Human breast cancer cells generated by oncogenic transformation of primary mammary epithelial cells.

By Brian Elenbaas, L. Spirio, F. Koerner, M. D. Fleming, D. Zimonjic, Joana Liu Donaher, N. Popescu, William C. Hahn, R. Weinberg. Genes & Development, 2001

936 citations recorded by the source corpus.

A number of genetic mutations have been identified in human breast cancers, yet the specific combinations of mutations required in concert to form breast carcinoma cells remain unknown. One approach to identifying the genetic and biochemical alterations required for this process involves the transformation of primary human mammary epithelial cells (HMECs) to carcinoma cells through the introduction of specific genes. Here we show that introduction of three genes encoding the SV40 large-T antigen, the telomerase catalytic subunit, and an H-Ras oncoprotein into primary HMECs results in cells that form tumors when transplanted subcutaneously or into the mammary glands of immunocompromised mice. The tumorigenicity of these transformed cells was dependent on the level of ras oncogene expression. Interestingly, transformation of HMECs but not two other human cell types was associated with amplifications of the c-myc oncogene, which occurred during the in vitro growth of the cells. Tumors derived from the transformed HMECs were poorly differentiated carcinomas that infiltrated through adjacent tissue. When these cells were injected subcutaneously, tumors formed in only half of the injections and with an average latency of 7.5 weeks. Mixing the epithelial tumor cells with Matrigel or primary human mammary fibroblasts substantially increased the efficiency of tumor formation and decreased the latency of tumor formation, demonstrating a significant influence of the stromal microenvironment on tumorigenicity. Thus, these observations establish an experimental system for elucidating both the genetic and cell biological requirements for the development of breast cancer.

Subfields: Biotechnology, Genetics, Oncology

View Paper: https://www.semanticscholar.org/paper/d2302faf38a20f929e5061fbf0d1521bcd0da57d

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Citations in the corpus

Poised chromatin at the ZEB1 promoter enables cell plasticity and enhances tumorigenicity (2013, neutral): “pBabe SV40-ER (Zeocin), pBabe H-Ras (Puromycin), PRRL-GFP, pLV-Tomato vectors, production of virus and infection of target cells have been previously described (Elenbaas et al., 2001; Shaner et al., 2004).”
The essence of senescence. (2010, background): “For instance, increased expression of activated RAS genes is a common feature of human tumors (Elenbaas et al. 2001; Johnson et al. 2005; Okamura et al. 2006; Chen et al. 2009) that can be associated with the conversion from benign to malignant tumors (Quintanilla et al. 1986; Finney and Bishop…”
Identification of conserved gene expression features between murine mammary carcinoma models and human breast tumors (2007, background): “In cultured human mammary epithelial cells, which show basal/myoepithelial characteristics [1,33], both high oncogenic H-ras and SV40 Large T-antigen expression are necessary for transformation [34].”
Alternative Isoform Regulation in Human Tissue Transcriptomes (2008, background): “By grouping tandem 39 UTRs by switch score, the most switch-like events showed increased sequence conservation relative to events with lower switch scores in the vicinity of and upstream of the proximal (59) polyadenylation signal (PAS), and also upstream of the distal (39) PAS (Fig.”
Collagen reorganization at the tumor-stromal interface facilitates local invasion (2006, background): “The importance of matrix composition and morphology around the mammary epithelium is illustrated by studies showing that misregulated stromal-epithelial interactions can promote tumorigenesis [6-8] and the fact that breast carcinomas often exhibit desmoplasia (excessive collagen surrounding an invas”
Generation of Breast Cancer Stem Cells through Epithelial-Mesenchymal Transition (2008, background): “Of note, the length of time necessary for the emergence of CD24(2) cells following HRas introduction is consistent with the initial observation by Elenbaas and colleagues that the tumorigenicity of HMLER cells was associated with the occurrence of secondary events [27].”
Direct Signaling Between Platelets and Cancer Cells Induces an Epithelial-Mesenchymal-Like Transition and Promotes Metastasis (2011, positive): “Weinberg) were cultivated as described previously (Elenbaas et al., 2001).”
Androgen-Induced Differentiation and Tumorigenicity of Human Prostate Epithelial Cells (2004, methodology): “PrECs were infected with combinations of amphotropic retroviruses encoding the SV40 large T antigen (LT), ST, hTERT, H-ras, phosphatidylinositol 3-kinase (PI3K), and MYC as described previously ( 16 , 21).”
Integrative genomic approaches identify IKBKE as a breast cancer oncogene. (2007, methodology): “Identification of IKBKE as a Human Breast Cancer Oncogene Mutations of proto-oncogenes and tumor suppressor genes allow incipient cancer cells to escape the normal mechanisms that regulate cell and tissue homeostasis.”
Identification of selective inhibitors of cancer stem cells by high-throughput screening (2009, methodology): “To this end, we modified experimentally transformed HMLER breast cancer cells (Elenbaas et al., 2001) by short hairpin RNA (shRNA)-mediated inhibition of the human CDH1 gene, which encodes E-cadherin.”
Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. (2008, methodology): “May 15, 2008 In consonance with previous findings ( 25 ), control HMLER cells did not form any macroscopic nodules in the lungs, and only a few GFP-positive cells could be detected as micrometastases in the lungs of animals bearing these tumors (Fig. 2D).”