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

Pandrangi SL, Raju Bagadi SA, Sinha NK, Kumar M, Dada R, Lakhanpal M, Soni A, Malvia S, Simon S, Chintamani C, Mohil RS, Bhatnagar D, Saxena S - Cancer Cell Int. (2014)

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India. sunita_saxena@yahoo.com.

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.

No MeSH data available.


Related in: MedlinePlus

Population doubling time: cells were plated in 6-well plates at a plating density of 1 × 105 cm2 in DMEM growth medium, supplemented with 10% FBS. Growth medium was renewed every 3 days. Cell counts were performed on the days indicated. Each point represents the mean of three different determinations made in triplicate. The population doubling times of the established cell lines (a) NIPBC-1 and (b) NIPBC-2 were calculated to be 33.25 hrs and 31.56 hrs respectively.
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Figure 9: Population doubling time: cells were plated in 6-well plates at a plating density of 1 × 105 cm2 in DMEM growth medium, supplemented with 10% FBS. Growth medium was renewed every 3 days. Cell counts were performed on the days indicated. Each point represents the mean of three different determinations made in triplicate. The population doubling times of the established cell lines (a) NIPBC-1 and (b) NIPBC-2 were calculated to be 33.25 hrs and 31.56 hrs respectively.

Mentions: Population doubling time of cell lines NIPBC-1 and NIPBC-2 cell lines were determined as described in materials and methods. The doubling time of NIPBC-1 cell line was found to be 33.25 hrs, while that of NIPBC-2 was 31.56 hrs (Figure 9).


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

Pandrangi SL, Raju Bagadi SA, Sinha NK, Kumar M, Dada R, Lakhanpal M, Soni A, Malvia S, Simon S, Chintamani C, Mohil RS, Bhatnagar D, Saxena S - Cancer Cell Int. (2014)

Population doubling time: cells were plated in 6-well plates at a plating density of 1 × 105 cm2 in DMEM growth medium, supplemented with 10% FBS. Growth medium was renewed every 3 days. Cell counts were performed on the days indicated. Each point represents the mean of three different determinations made in triplicate. The population doubling times of the established cell lines (a) NIPBC-1 and (b) NIPBC-2 were calculated to be 33.25 hrs and 31.56 hrs respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4016554&req=5

Figure 9: Population doubling time: cells were plated in 6-well plates at a plating density of 1 × 105 cm2 in DMEM growth medium, supplemented with 10% FBS. Growth medium was renewed every 3 days. Cell counts were performed on the days indicated. Each point represents the mean of three different determinations made in triplicate. The population doubling times of the established cell lines (a) NIPBC-1 and (b) NIPBC-2 were calculated to be 33.25 hrs and 31.56 hrs respectively.
Mentions: Population doubling time of cell lines NIPBC-1 and NIPBC-2 cell lines were determined as described in materials and methods. The doubling time of NIPBC-1 cell line was found to be 33.25 hrs, while that of NIPBC-2 was 31.56 hrs (Figure 9).

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi 110029, India. sunita_saxena@yahoo.com.

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.

No MeSH data available.


Related in: MedlinePlus