Table 2.

Fold Change in Gene Expression in MEWO Melanoma Cells at 48 Hours (before Extensive Apoptosis) and 96 Hours (after Completion of the Apoptotic Response) following the Transition from a Methionine-Efficient State to the MDS State as Determined by Reverse Transcription-PCR Analysis

48 h96 h
Effector genes
    Loss of proliferative activity
    Apoptosis related
Mediator genes
    Cell cycle/mitosis inhibitors
        CDKN1A (p21)2329
        CDKN1B (p27Kip1)13
        CDKN1C (p57Kip2)253
        CDKN2B (p15)−3−110
        CDKN2C (p18)−7−16
        CDKN2D (p19)1−6
        Cyclin B2−3
        Aurora kinase B−2−13
    Apoptosis related
Resistance-related genes
  • NOTE: Effector genes were selected based on their high intensity of expression under MDS as determined by cDNA oligoarray analysis. Mediator genes were selected based on their high intensity of expression and their likelihood to be involved in the physiologic changes observed in response to MDS. Genes involved in the resistance to chemotherapy/radiation were selected according to similar criteria.

  • * Amplification was carried for a maximum of 30 cycles. Three serial dilutions of control and MDS samples having equal levels of β-actin were amplified and runs were analyzed with light cycle software under conditions of a low error rate (<0.2). Slopes determined from plots of cDNA concentration versus cycles required to achieve 30% of the maximum SYBR green fluorescence (Santa Cruz Biotechnology) were used to relative to β-actin expression of each gene in MDS and control samples. Fold increase (positive) or fold decrease (negative) in comparison with methionine controls were determined from ratios of MDS to control relative RNA concentrations under equal β-actin levels.