Technical Reference #1751

1751.

Glyceraldehyde 3-Phosphate Dehydrogenase Depletion Induces Cell Cycle Arrest and Resistance to Antimetabolites in Human Carcinoma Cell Lines Manali S. Phadke; Natalia F. Krynetskaia; Anurag K. Mishra; Evgeny Krynetskiy, Temple University, Journal of Pharmacology and Experimental Therapeutics, 331(1751), (2009)
Link To Paper

Materials & Methods:
Cell Cultures Drug Treatment and Plasmids. Lung carcinoma cells A549 and NCI-H358 were obtained from the American Type Culture Collection (Manassas VA) and renal carcinoma UO31 cells were obtained from the Tumor Cell Line Repository National Cancer Institute (Frederick MD). A549 cells were maintained in Ham’s F-12K medium and NCI-H358 and UO31 cells were maintained in RPMI 1640 medium (American Type Culture Collection) at 40 to 80% confluence. Cells were treated with drugs dissolved in 0.1 N NaOH (mercaptopurine MP) or water (cytarabine araC) as 500 to 1000 stock solutions; drug concentrations were determined spectrophotometrically (MP ε320 19600; araC ε272 9259) (O’Neil et al. 2001). [5-3H]Cytosine- -D-arabinofuranoside (14.9 Ci/mmol) (Moravek Biochemicals Brea CA) was used in DNA incorporation experiments (Krynetskaia et al. 2009). Human GAPDH cDNA was inserted in frame with EGFP into pcDNA3.1 as described previously (Brown et al. 2004) and verified by sequencing.

Microscopic Technique
Confocal Microscopy

Cell Type(s)
Lung carcinoma cells A549 & NCI-H358; UO31 cells