Establishment and characterisation of two novel breast cancer cell lines

Abstract

Two novel oestrogen receptor (ER) negative breast cancer cell lines, BCa-11 (familial) and BCa-15 (sporadic) were successfully established from primary tumours. Characterisation of these cell lines showed expression of epithelial specific antigen and cytokeratins confirming their epithelial lineage. Analysis of ultrastructure and anchorage independent growth confirmed the epithelial nature and transformed phenotype of these cells. Both cell lines showed loss of pRb, Dab2 and ERalpha and elevated levels of proliferation marker Ki67. In addition, BCa-11 cells showed loss of HOXA5, tumour suppressor genes p16(INK4A) and RARbeta as well as overexpression of CyclinD1. Elevation of DNMT1 and DNMT3B transcript levels, promoter hypermethylation of RASSF1A, RARbeta2, and HOXA5 further support their neoplastic origin. In conclusion, the two ERalpha negative breast cancer cell lines established herein have certain useful characteristics that may make them valuable for understanding the mechanism of oestrogen receptor negative breast tumours and testing new drugs.

Establishment and characterization of two primary breast cancer cell lines from young Indian breast cancer patients: mutation analysis

“…Although a number of normal and breast cancer cell lines established from western patients are available, there is a paucity of cell lines established from Indian breast cancer patients. So far only two breast cancer cell lines have been established from late onset breast cancer patients (>60 years) using primary tumors [24], hence the two cell lines we have established in the present study are unique with respect to the ethnicity also as these are the first breast cancer cell lines to be established from Indian continent representing early onset breast cancers. In conclusion, in the present study we have established two novel triple negative breast cancer cell lines, NIPBC-1 and NIPBC-2 from primary tumors of two early onset breast cancers (<40 years), which may serve as valuable in vitro models to study breast tumorigenesis in young breast cancer patients and identification of unique therapeutic targets.…”

“…Majority of breast cancer derived cell lines are from secondary tumours and pleural effusions of patients with advanced stage breast cancers [10-18]. Few breast cancer cell lines have been successfully established from primary tumours [19-24]. Among the available breast cancer cell lines, majority of cell lines are established from cancers from older age group patients (> 55 yrs), only a small proportion of breast cancer cell lines are established from patients <40 years of age.…”

A sensitive test for the detection of specific DSB repair defects in primary cells from breast cancer specimens

“…Clinical staging followed the classification of American Joint Committee on Cancer revised 6th edition 22. Breast carcinoma tissues were collected and processed for setting up the primary cell cultures as detailed in Bagadi et al 23. Enrichment of epithelial cells was achieved by differential trypsinization as described previously 23, 24.…”

“…Breast carcinoma tissues were collected and processed for setting up the primary cell cultures as detailed in Bagadi et al 23. Enrichment of epithelial cells was achieved by differential trypsinization as described previously 23, 24. Thereafter fibroblast overgrowth was reduced by selective chemical elimination using geneticin (G418 sulfate, 100 μg/ml; GIBCO BRL, Grand Island, NY) as described elsewhere 25.…”

“…Cells grown on coverslips were fixed with acetone and processed for immunocytochemical analysis as described in Bagadi et al 23. Briefly, cells were immunolabeled with primary antibodies (1 μg/ml) followed by biotinylated secondary antibody, (0.8 μg/ml) (DAKO, Produktionsvej, Glostrup, Denmark).…”