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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

Representative metaphases (a, b) of NIPBC-2 cells, at passages 15 and 52, with trypsin-giemsa banding. Karyotypes (c, d) of the above metaphases showing near tetraploidy with a modal number of 58 to 62 chromosomes.
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Figure 11: Representative metaphases (a, b) of NIPBC-2 cells, at passages 15 and 52, with trypsin-giemsa banding. Karyotypes (c, d) of the above metaphases showing near tetraploidy with a modal number of 58 to 62 chromosomes.

Mentions: Karyotype analysis of NIPBC-1 and NIPBC-2 cells has shown that both the cell lines possess aneuploidy. Chromosomes 7, 9, X and 11 showed deletions in various regions in both the cell lines. Cytogenetic analysis has shown multiple rearrangements. NIPBC-1 was near tetraploid with a modal number of 58 to 62 chromosomes, most of the chromosomes exhibited several translocations and marker chromosomes; rearrangements like t(14:15) (q12:q12) and i(17q) were found commonly in these cells; Isochromosomes 17q was the most common aberration identified in NIPBC-1 (Figure 8). NIPBC-2 cell line was also found to be aneuploid with nearly tetraploid to pentaploid complement and the chromosomal numbers ranged from 107 to110 (Figures 10 and 11). No significant karyotype changes were found in the karyotypic analysis among the early and late passages of both the cell lines; indicating that both NIPBC-1 and NIPBC-2 are stable cell lines.


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)

Representative metaphases (a, b) of NIPBC-2 cells, at passages 15 and 52, with trypsin-giemsa banding. Karyotypes (c, d) of the above metaphases showing near tetraploidy with a modal number of 58 to 62 chromosomes.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 11: Representative metaphases (a, b) of NIPBC-2 cells, at passages 15 and 52, with trypsin-giemsa banding. Karyotypes (c, d) of the above metaphases showing near tetraploidy with a modal number of 58 to 62 chromosomes.
Mentions: Karyotype analysis of NIPBC-1 and NIPBC-2 cells has shown that both the cell lines possess aneuploidy. Chromosomes 7, 9, X and 11 showed deletions in various regions in both the cell lines. Cytogenetic analysis has shown multiple rearrangements. NIPBC-1 was near tetraploid with a modal number of 58 to 62 chromosomes, most of the chromosomes exhibited several translocations and marker chromosomes; rearrangements like t(14:15) (q12:q12) and i(17q) were found commonly in these cells; Isochromosomes 17q was the most common aberration identified in NIPBC-1 (Figure 8). NIPBC-2 cell line was also found to be aneuploid with nearly tetraploid to pentaploid complement and the chromosomal numbers ranged from 107 to110 (Figures 10 and 11). No significant karyotype changes were found in the karyotypic analysis among the early and late passages of both the cell lines; indicating that both NIPBC-1 and NIPBC-2 are stable cell lines.

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