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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.


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Representative pictures of mammospheres formed by NIPBC1 & NIPBC-2 cell lines. (Magnification- 40x, inset mag- 100x).
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Figure 17: Representative pictures of mammospheres formed by NIPBC1 & NIPBC-2 cell lines. (Magnification- 40x, inset mag- 100x).

Mentions: CSCs have been isolated from many solid tumors, including breast [36,79], pancreatic [80,81], brain [82,83], colon [84-86], liver [87], head/neck [88], ovarian [89,90], and melanoma [91,92]. CSCs were first isolated and characterized by Al-Hajj et al. using the cell surface marker CD44+/CD24−/low/Lin − and recapitulated the heterogeneity of the original tumor by injecting them into nude mice [36]. Both NIPBC-1 and NIPBC-2 cell lines showed the presence of 0.2% and 0.1% of CD44+/CD24- breast cancer stem cells respectively which is relatively similar to the commercially available breast cancer cell line. When plated in serum free, non-adherent conditions as described earlier [33], both NIPBC-1 and NIPBC-2 cell lines formed mammospheres (Figure 17) and could be further passaged for three generations.


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 pictures of mammospheres formed by NIPBC1 & NIPBC-2 cell lines. (Magnification- 40x, inset mag- 100x).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 17: Representative pictures of mammospheres formed by NIPBC1 & NIPBC-2 cell lines. (Magnification- 40x, inset mag- 100x).
Mentions: CSCs have been isolated from many solid tumors, including breast [36,79], pancreatic [80,81], brain [82,83], colon [84-86], liver [87], head/neck [88], ovarian [89,90], and melanoma [91,92]. CSCs were first isolated and characterized by Al-Hajj et al. using the cell surface marker CD44+/CD24−/low/Lin − and recapitulated the heterogeneity of the original tumor by injecting them into nude mice [36]. Both NIPBC-1 and NIPBC-2 cell lines showed the presence of 0.2% and 0.1% of CD44+/CD24- breast cancer stem cells respectively which is relatively similar to the commercially available breast cancer cell line. When plated in serum free, non-adherent conditions as described earlier [33], both NIPBC-1 and NIPBC-2 cell lines formed mammospheres (Figure 17) and could be further passaged for three generations.

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