Molecular and cellular characterization of two patient-derived ductal carcinoma in situ (DCIS) cell lines, ETCC-006 and ETCC-010

Abstract

Background Currently it is unclear how in situ breast cancer progresses to invasive disease; therefore, a better understanding of the events that occur during the transition to invasive carcinoma is warranted. Here we have conducted a detailed molecular and cellular characterization of two, patient-derived, ductal carcinoma in situ (DCIS) cell lines, ETCC-006 and ETCC-010. Methods Human DCIS cell lines, ETCC-006 and ETCC-010, were compared against a panel of cell lines including the immortalized, breast epithelial cell line, MCF10A, breast cancer cell lines, MCF7 and MDA-MB-231, and another DCIS line, MCF10DCIS.com. Cell morphology, hormone and HER2/ERBB2 receptor status, cell proliferation, survival, migration, anchorage-independent growth, indicators of EMT, cell signalling pathways and cell cycle proteins were examined using immunostaining, immunoblots, and quantitative, reverse transcriptase PCR (qRT-PCR), along with clonogenic, wound-closure and soft agar assays. RNA sequencing (RNAseq) was used to provide a transcriptomic profile. Results ETCC-006 and ETCC-010 cells displayed notable differences to another DCIS cell line, MCF10DCIS.com, in terms of morphology, steroid-receptor/HER status and markers of EMT. The ETCC cell lines lack ER/PR and HER, form colonies in clonogenic assays, have migratory capacity and are capable of anchorage-independent growth. Despite being isogenic, less than 30% of differentially expressed transcripts overlapped between the two lines, with enrichment in pathways involving receptor tyrosine kinases and DNA replication/cell cycle programs and in gene sets responsible for extracellular matrix organisation and ion transport. Conclusions For the first time, we provide a molecular and cellular characterization of two, patient-derived DCIS cell lines, ETCC-006 and ETCC-010, facilitating future investigations into the molecular basis of DCIS to invasive ductal carcinoma transition.

Multicellular spheroids containing synthetic mineral particles: an advanced 3D tumor model system to investigate breast precancer malignancy potential according to the mineral type

“…These cells are derived from MCF10A cells, a normal human epithelial cell line isolated from a patient with fibrocystic breasts, 41 and are estrogen receptor (ER) negative as well as progestogen receptor (PR) and human epidermal growth factor receptor (Her2) positive. 42 MCF10DCIS.com cells were shown to spontaneously deposit MC-like mineral particles after ten days of culture as multicellular spheroids. 43 In our model, we did not wait for spontaneous mineral deposition.…”

Microcalcifications can either trigger or suppress breast precancer malignancy potential according to the mineral type in a 3D tumor model

“…Specifically, we used precancer cells of the human cell line MCF10DCIS.com, which, in vivo , induce DCIS. These cells are derived from MCF10A cells, a normal human epithelial cell line isolated from a patient with fibrocystic breasts 33 and are estrogen receptor (ER) negative as well as progestogen receptor (PR) and human epidermal growth factor receptor (Her2) positive 34 . MCF10DCIS.com cells were shown to spontaneously deposit MC-like mineral particles after ten days of culture as multicellular spheroids 35 .…”