Characterizing Cisplatin-Resistant Ovarian Cancer

OVCA429 cells were isolated from a patient with late stage, cisplatin-resistant ovarian carcinoma. OVCA429/pCEG cells carrying a green fluorescence protein empty vector and OVCA429/NICD3 cells constitutively expressing green fluorescence protein tagged with NICD3 were sorted by a fluorescence-activated cell sorter and maintained in RPMI 1640 medium (Mediatech Inc, Herndon, VA) supplemented with 10% heat-inactivated fetal bovine serum and 100 U/mL penicillin and streptomycin at 37°C in a 5% CO₂ incubator [34] (link). N-acetyl cysteine (NAC) and MSeA (Sigma-Aldrich, St. Louis, MO) were dissolved in phosphate-buffered saline. NAC is an antioxidant that mainly abolishes hydrogen peroxide. Carboplatin (Enzo Life Sciences, Farmingdale, NY) was dissolved in water. KU 60019 and NU 7026 (Tocris, Ellisville, MO) were dissolved in dimethyl sulfoxide, and were ATP-competitive, selective chemical inhibitors of the kinase activity of ATM and DNA-PK_cs[35] (link), respectively.

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Tzeng T.J., Cao L., Fu Y., Zeng H, & Cheng W.H. (2014). Methylseleninic Acid Sensitizes Notch3-Activated OVCA429 Ovarian Cancer Cells to Carboplatin. PLoS ONE, 9(7), e101664.

Publication 2014

RPMI 1640 medium N-acetyl cysteine (NAC) Carboplatin KU 60019 NU 7026

Antioxidant Carboplatin Cell Chemical activity Cisplatin Dimethyl sulfoxide Dna pkcs Fetal bovine serum Fluorescence Green fluorescence protein Hydrogen peroxide Inhibitors Kinase Ku 60019 N acetyl cysteine Nu 7026 Ovarian carcinoma Patient Penicillin Phosphate Saline Streptomycin Vector

Corresponding Organization : Grand Forks Human Nutrition Research Center

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Variable analysis

independent variables

N-acetyl cysteine (NAC)
Carboplatin
KU 60019
NU 7026

dependent variables

Cell viability/proliferation
Cellular response to treatments

control variables

OVCA429/pCEG cells carrying a green fluorescence protein empty vector
OVCA429/NICD3 cells constitutively expressing green fluorescence protein tagged with NICD3
RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum and 100 U/mL penicillin and streptomycin
Incubation at 37°C in a 5% CO2 incubator

controls

Positive control: OVCA429/NICD3 cells constitutively expressing green fluorescence protein tagged with NICD3
Negative control: OVCA429/pCEG cells carrying a green fluorescence protein empty vector

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