UACC-732 (ATCC® CRL-3166)

Organism: Homo sapiens, human  /  Tissue: breast  /  Disease: inflammatory carcinoma of the breast

Organism Homo sapiens, human
Tissue breast
Product Format frozen
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 1
Disease inflammatory carcinoma of the breast
Age 33 years old
Gender female
Ethnicity Caucasian
Applications
The cells have been used for evaluating drug resistance to HER-2 inhibitors, such as trastuzumab and lapatinib. In addition, UACC-732 has been shown to be a drug resistant cell line to cyclin D kinase 4/6 inhibitor and HER-2 inhibitors.
Storage Conditions liquid nitrogen vapor phase
Karyotype range=38-76; modal number=71-75
Images
Derivation This human breast cancer cell line, UACC-732, was derived from a 33 year-old female with infiltrating ductal inflammatory carcinoma of the breast metastatic to the pleural fluid. Pathology confirmed that the specimen was a metastatic adenocarcinoma consistent with inflammatory carcinoma of the breast.
Receptor Expression
progesterone receptor (PR), expressed
epidermal growth factor receptor (EGFR), low expression
estrogen receptor (ER), not expressed
Oncogene HER2/neu, overexpressed
Comments
This cell line expresses receptor HER-2/neu.

Complete Growth Medium

The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium:

  • fetal bovine serum to a final concentration of 5%
  • 0.01 mg/ml transferrin (final conc.)
  • 0.01 mg/ml insulin (final conc.)
  • 5 µg/mL (55 U/ml) catalase (final conc.)
  • 3.6 µg/mL (0.01 mM) hydrocortisone (final conc.)
  • 70 µg/mL (0.5mM) o-phosphoethanolamine (final conc.)
  • 10 ng/ml human recombinant epidermal growth factor (EGF) (final conc.)
  • 3 ng/ml (0.01 µM) estradiol (final conc.)
  • 0.8 ng/ml (1 pM) Na-L-thyroxine (final conc.)
  • 23 µg/mL (0.2mM) proline (final conc.)
  • extra 2 mM glutamine
Note: Do not filter complete medium.
L-15 Medium is formulated for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation.
Subculturing
These cells are cultured on collagen I coated vessels. Add 5 µg per cm2 collagen I (BD Biosciences, Cat. No.354236 or equivalent) to culture vessels and incubate at room temperature for 1 hour. Remove collagen solution and rinse vessels 3 times with a balanced salt solution. Vessels may be used immediately or air dried and stored at 2-8°C for up to one week under sterile conditions. Alternatively, commercially available pre-coated Collagen I vessels, such as BD BioCoat Cellware (BD Biosciences, Cat. No. 356524 for 75 cm2 flask) or equivalent, may be used.
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of solutions for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (DPBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new collagen I coated culture vessels. An inoculum of 3 X 104 to 4 X 104 viable cells/cm2 is recommended.
  7. Incubate cultures at 37.0°C.
Subculture when cultures reach a cell concentration between 7 X 104 to 1 X 105
Subcultivation ratio: A subcultivation ratio of 1:2 is recommended.
Medium renewal: every 2 to 3 days
Cryopreservation
Freeze medium: complete growth medium, 80%; FBS, 10%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37.0°C
Atmosphere: air, 100%
Growth Conditions: This cell line grows slowly.
STR Profile
D5S818: 13, 14
D13S317: 10, 12
D7S820: 12
D16S539: 9, 10
vWA: 16,19
TH01: 6, 9.3
Amelogenin:X
TPOX: 8, 11
CSF1PO: 10,12
Name of Depositor K Brown
References

Domann FE, et al. Epigenetic silencing of maspin gene expression in human breast cancers. Int. J. Cancer. 85(6): 805-810, 2000 PubMed: 10709100

Kauraniemi P, et al. New amplified and highly expressed genes discovered in the ERBB2 amplicon in breast cancer by cDNA microarrays. Cancer Res. 61(22): 8235-8240, 2001 PubMed: 117194555

Ortiz RM, et al. Aberrant alternative exon use and increased copy number of human metalloprotease-disintegrin ADAM15 gene in breast cancer cells. Genes Chromosomes Cancer. 41(4): 366-378, 2004. PubMed: 15384173

Futscher BW, et al. Chapter 6: Epigenetic Dysregulation of Maspin (SerpinB5) in Cancer Invasion and Metastasis. DNA Methylation, Epigenetics and Metastasis. Cancer Metastasis - Biology and Treatment, 7: 133-155, 2005.

Oshiro MM, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 7(5): R669-R680, 2005. PubMed: 16168112

Alarmo EL, et al. Bone morphogenetic protein 7 is widely overexpressed in primary breast cancer. Genes Chromosomes Cancer. 45(4): 411-419, 2006. PubMed: 16419056

Futscher BW, et al. Chapter 7: Epigenetic Regulation of Genes That Affect Tumor Cell Adhesion. Cell Adhesion and Cytoskeletal Molecules in Metastasis. Cancer Metastasis - Biology and Treatment, 9: 123-140, 2006.

Alarmo EL, et al. A comprehensive expression survey of bone morphogenetic proteins in breast cancer highlights the importance of BMP4 and BMP7. Breast Cancer Res. Treat. 103(2): 239-246, 2007. PubMed: 17004110

Finn RS, et al. Dasatinib, an orally active small molecule inhibitor of both the src and abl kinases, selectively inhibits growth of basal-type/"triple-negative" breast cancer cell lines growing in vitro. Breast Cancer Res. Treat. 105(3): 319-326, 2007. PubMed: 17268817

Robey IF, et al. Regulation of the warburg effect in early-passage breast cancer cells. Neoplasia 10(8): 745-756, 2008. PubMed: 18670636

Parssinen J, et al. PPM1D silencing by RNA interference inhibits proliferation and induces apoptosis in breast cancer cell lines with wild-type p53. Cancer Genet. Cytogenet. 182(1): 33-39, 2008. PubMed: 18328948

Finn RS, et al. PD 0332991, a selective cyclin D kinase 4/6 inhibitor, preferentially inhibits proliferation of luminal estrogen receptor-positive human breast cancer cell lines in vitro. Breast Cancer Res. 11(5): R77, 2009. PubMed: 19874578

Ketolainen JM, et al. Parallel inhibition of cell growth and induction of cell migration and invasion in breast cancer cells by bone morphogenetic protein 4. Breast Cancer Res. Treat. 124(2): 377-386, 2010. PubMed: 20182795

O'Brien NA, et al. Activated phosphoinositide 3-kinase/AKT signaling confers resistance to trastuzumab but not lapatinib. Mol. Cancer Ther. 9(6): 1489-1502, 2010. PubMed: 20501798

Basic Documentation
Other Documentation
References

Domann FE, et al. Epigenetic silencing of maspin gene expression in human breast cancers. Int. J. Cancer. 85(6): 805-810, 2000 PubMed: 10709100

Kauraniemi P, et al. New amplified and highly expressed genes discovered in the ERBB2 amplicon in breast cancer by cDNA microarrays. Cancer Res. 61(22): 8235-8240, 2001 PubMed: 117194555

Ortiz RM, et al. Aberrant alternative exon use and increased copy number of human metalloprotease-disintegrin ADAM15 gene in breast cancer cells. Genes Chromosomes Cancer. 41(4): 366-378, 2004. PubMed: 15384173

Futscher BW, et al. Chapter 6: Epigenetic Dysregulation of Maspin (SerpinB5) in Cancer Invasion and Metastasis. DNA Methylation, Epigenetics and Metastasis. Cancer Metastasis - Biology and Treatment, 7: 133-155, 2005.

Oshiro MM, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 7(5): R669-R680, 2005. PubMed: 16168112

Alarmo EL, et al. Bone morphogenetic protein 7 is widely overexpressed in primary breast cancer. Genes Chromosomes Cancer. 45(4): 411-419, 2006. PubMed: 16419056

Futscher BW, et al. Chapter 7: Epigenetic Regulation of Genes That Affect Tumor Cell Adhesion. Cell Adhesion and Cytoskeletal Molecules in Metastasis. Cancer Metastasis - Biology and Treatment, 9: 123-140, 2006.

Alarmo EL, et al. A comprehensive expression survey of bone morphogenetic proteins in breast cancer highlights the importance of BMP4 and BMP7. Breast Cancer Res. Treat. 103(2): 239-246, 2007. PubMed: 17004110

Finn RS, et al. Dasatinib, an orally active small molecule inhibitor of both the src and abl kinases, selectively inhibits growth of basal-type/"triple-negative" breast cancer cell lines growing in vitro. Breast Cancer Res. Treat. 105(3): 319-326, 2007. PubMed: 17268817

Robey IF, et al. Regulation of the warburg effect in early-passage breast cancer cells. Neoplasia 10(8): 745-756, 2008. PubMed: 18670636

Parssinen J, et al. PPM1D silencing by RNA interference inhibits proliferation and induces apoptosis in breast cancer cell lines with wild-type p53. Cancer Genet. Cytogenet. 182(1): 33-39, 2008. PubMed: 18328948

Finn RS, et al. PD 0332991, a selective cyclin D kinase 4/6 inhibitor, preferentially inhibits proliferation of luminal estrogen receptor-positive human breast cancer cell lines in vitro. Breast Cancer Res. 11(5): R77, 2009. PubMed: 19874578

Ketolainen JM, et al. Parallel inhibition of cell growth and induction of cell migration and invasion in breast cancer cells by bone morphogenetic protein 4. Breast Cancer Res. Treat. 124(2): 377-386, 2010. PubMed: 20182795

O'Brien NA, et al. Activated phosphoinositide 3-kinase/AKT signaling confers resistance to trastuzumab but not lapatinib. Mol. Cancer Ther. 9(6): 1489-1502, 2010. PubMed: 20501798