- References (2)
- Inventor Info
|Antigen/Gene or Protein Targets||BCAR1, BCAR2|
|Disease Keywords||Mammary ductal carcinoma, estrogen independent|
Endocrine therapy of breast cancer has been applied widely and proven to be effective. However, in many instances endocrine treatments ultimately fail due to the development of an estrogen-independent therapy resistant phenotype. To elucidate the molecular mechanism underlying this endocrine therapy failure, the laboratory of Lambert Dorssers applied random insertional mutagenesis using defective retroviruses to identify the main genes conferring estrogen independence.
Out of 15 candidate BCAR genes, several including BCAR1 and BCAR2 were shown to directly underlie estrogen independence. Somatic cell fusions were generated for these two genes in the hygromycin-B-resistant variant of ZR-75-1 breast cancer cells (ZH3D7) resulting in a panel of 3 cell lines (Cat No 154639-154641).
These cell lines are a powerful tool for studying the molecular and cellular mechanisms of breast tumour progression and therapy resistance.
|Production Details||Hygromycin-B-resistant variants of ZR-75-1 cells, ZH3D7, were used as recipients in the somatic-cell-fusion experiments. Donor cells were two different anti-estrogen-resistant cell lines. Approximately 6 million donor cells, which were gamma-irradiated with approx 40 Gy and 3 million recipient cells were plated in 25cm flasks in RBCS medium with estradiol. After strong adherence to the flasks in 36 to 48 hr, cells were washed 3 times in RPMI-1640 without serum and incubated with 1 ml polyethylene glycol (PEG) 1500 at 30–34°C for 1 min on a stretching table. The PEG solution was diluted with1, 2 and 4 ml RPMI-1640 after 1, 2 and 4 min respectively. Finally,cells were washed 3 times with RPMI-1640 and incubated for 2days at 37°C in RBCS medium with estradiol. Selection with 1mg/ml G418 in RBCS medium plus estradiol was performed for one week followed by one week of selection with 50 µg/ml hygromycin B. After an additional week of selection with G418, colonies were picked and expanded to cell lines in RBCS medium containing estradiol and G418|
|Research Area||Cancer, Drug Discovery & Development|
|Recommended Growing Conditions||RBCS medium containing estradiol and Geneticin (G418)|
|Notes||These cell lines are resistant to hygromycin and Geneticin and are maintained in RBCS medium with estradiol and Geneticin. Since they carry a BCAR gene, that can also proliferate slowly in medium without estradiol and supplemented with anti-estrogen.|
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