Journal
BMC CANCER
Volume 21, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s12885-021-08511-2
Keywords
Ductal carcinoma in situ (DCIS); Breast carcinoma cell lines; qRT-PCR; RNA sequencing (RNAseq); Proliferation; Migration; Anchorage-independent growth; Epithelial to mesenchymal transition; Cell signalling pathways; Cell cycle
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Funding
- Translational Research Access Programme (TRAP) Award from the School of Medicine at University College Cork
- Boehringer Ingelheim
- Collaborative Research Centre 992 Medical Epigenetics (DFG grant) [SFB 992/1 2012]
- German Federal Ministry of Education and Research (BMBF) [031 A538A/A538C RBC, 031L0101B/031L0101C de. NBI-epi, 031 L0106 de. STAIR (de. NBI)]
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This study conducted a molecular and cellular characterization of two patient-derived DCIS cell lines, ETCC-006 and ETCC-010, revealing differences in migration and anchorage-independent growth capabilities compared to other DCIS cell lines. 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.
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.
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