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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

Invasion assay of NIPBC-1 and NIPBC-2 cell lines. Representative pictures of (a) MDA MB 231 (positive control), (b) NIPBC-1 and (c) NIPBC-2 cells invaded through Matrigel. (d) Cell number quantification of invasion.
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Figure 12: Invasion assay of NIPBC-1 and NIPBC-2 cell lines. Representative pictures of (a) MDA MB 231 (positive control), (b) NIPBC-1 and (c) NIPBC-2 cells invaded through Matrigel. (d) Cell number quantification of invasion.

Mentions: To examine the invasion capacity of the two cell lines established in the present study, we have carried out invasion assay on matrigel coated membrane inserts. We found invaded cells on the other side of the membrane upon overnight incubation with chemo attractant (FBS). Breast cancer cell line MDA-MB231 was taken as positive control. The number of cells invading the matrigel matrix are counted in atleast 4 randomly choosen fields per well. The numbers of MDA-MB231, NIPBC-1 and NIPBC-2 cells that invaded through the basement membrane were 527 ± 45.9020, 409.3 ± 32.3161, 290.6 ± 44.2417 respectively (Figure 12a-c). The number of NIPBC-1 cells that invaded through the basement membrane was significantly higher than that of NIPBC-2 cell line (p = 0.0407) (Figure 12d).


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)

Invasion assay of NIPBC-1 and NIPBC-2 cell lines. Representative pictures of (a) MDA MB 231 (positive control), (b) NIPBC-1 and (c) NIPBC-2 cells invaded through Matrigel. (d) Cell number quantification of invasion.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 12: Invasion assay of NIPBC-1 and NIPBC-2 cell lines. Representative pictures of (a) MDA MB 231 (positive control), (b) NIPBC-1 and (c) NIPBC-2 cells invaded through Matrigel. (d) Cell number quantification of invasion.
Mentions: To examine the invasion capacity of the two cell lines established in the present study, we have carried out invasion assay on matrigel coated membrane inserts. We found invaded cells on the other side of the membrane upon overnight incubation with chemo attractant (FBS). Breast cancer cell line MDA-MB231 was taken as positive control. The number of cells invading the matrigel matrix are counted in atleast 4 randomly choosen fields per well. The numbers of MDA-MB231, NIPBC-1 and NIPBC-2 cells that invaded through the basement membrane were 527 ± 45.9020, 409.3 ± 32.3161, 290.6 ± 44.2417 respectively (Figure 12a-c). The number of NIPBC-1 cells that invaded through the basement membrane was significantly higher than that of NIPBC-2 cell line (p = 0.0407) (Figure 12d).

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