title: Establishment and characterization of two primary breast cancer cell lines from young Indian breast cancer patients: mutation analysis.
abstract: Two novel triple negative breast cancer cell lines, NIPBC-1 and NIPBC-2 were successfully established from primary tumors of two young breast cancer patients aged 39 and 38 years respectively, diagnosed as infiltrating duct carcinoma of breast. Characterization of these cell lines showed luminal origin with expression of epithelial specific antigen and cytokeratin 18 and presence of microfilaments and secretary vesicles, microvilli, tight junctions and desmosomes on ultra-structural analysis. Both the cell lines showed anchorage independent growth and invasion of matrigel coated membranes. Karyotype analysis showed aneuploidy, deletions and multiple rearrangements in chromosomes 7, 9, X and 11 and isochromosomes 17q in both the cell lines. P53 mutational analysis revealed no mutation in the coding region in both the cell lines; however NIPBC-2 cell line showed presence of heterozygous C/G polymorphism, g.417 C > G (NM_000546.5) resulting in Arg/Pro allele at codon 72 of exon 4. Screening for mutations in BRCA1&2 genes revealed presence of three heterozygous polymorphisms in exon 11 of BRCA1 and 2 polymorphisms in exons 11, and14 of BRCA2 gene in both the cell lines. Both the cell lines showed presence of CD 44+/24-breast cancer stem cells and capability of producing mammosphere on culture. The two triple negative breast cancer cell lines established from early onset breast tumors can serve as novel invitro models to study mechanisms underlying breast tumorigenesis in younger age group patients and also identification of new therapeutic modalities targeting cancer stem cells.
authors: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Santhi Latha
authors: SL
authors: Pandrangi
authors: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Sarangadhara Appala
authors: SA
authors: Raju Bagadi
authors: Rajiv Gandhi Cancer Institute and Research Centre, Rohini, New Delhi, India.
authors: Navin Kumar
authors: NK
authors: Sinha
authors: Lab for Molecular Reproduction and Genetics, Anatomy department, All India Institute of medical Sciences, New Delhi, India.
authors: Manoj
authors: M
authors: Kumar
authors: Lab for Molecular Reproduction and Genetics, Anatomy department, All India Institute of medical Sciences, New Delhi, India.
authors: Rima
authors: R
authors: Dada
authors: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Meena
authors: M
authors: Lakhanpal
authors: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Abha
authors: A
authors: Soni
authors: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Shreshtha
authors: S
authors: Malvia
authors: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Sheeba
authors: S
authors: Simon
authors: Department of Surgery, Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Chintamani
authors: C
authors: Chintamani
authors: Department of Surgery, Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Ravindar Singh
authors: RS
authors: Mohil
authors: Department of Surgery, Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Dinesh
authors: D
authors: Bhatnagar
authors: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India.
authors: Sunita
authors: S
authors: Saxena