Abstract
Human malignant melanoma cell line UACC903 is resistant to apoptosis while chromosome 6-mediated suppressed cell line UACC903(+6) is sensitive. Here, we describe identification of differential molecular pathways underlying this difference. Using our recently developed mitochondria-focused cDNA microarrays, we identified 154 differentially expressed genes including proapoptotic (BAK1 [6p21.3], BCAP31, BNIP1, CASP3, CASP6, FAS, FDX1, FDXR, TNFSF10 and VDAC1) and antiapoptotic (BCL2L1, CLN3 and MCL1) genes. Expression of these pro- and anti-apoptotic genes was higher in UACC903(+6) than in UACC903 before UV treatment and was altered after UV treatment. qRT-PCR and Western blots validated microarray results. Our bioinformatic analysis mapped these genes to differential molecular pathways that predict resistance and sensitivity of UACC903 and UACC903(+6) to apoptosis respectively. The pathways were functionally confirmed by the FAS ligand-induced cell death and by siRNA knockdown of BAK1 protein. These results demonstrated the differential molecular pathways underlying survival and apoptosis of UACC903 and UACC903(+6) cell lines.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Apoptosis / drug effects
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Apoptosis / genetics*
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Apoptosis / radiation effects
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Apoptosis Regulatory Proteins / genetics*
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Cell Line, Tumor
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Cell Survival / drug effects
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Cell Survival / genetics
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Cell Survival / radiation effects
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Computational Biology / methods
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DNA, Mitochondrial / analysis
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DNA, Mitochondrial / genetics*
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DNA, Mitochondrial / radiation effects
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Fas Ligand Protein / metabolism
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Fas Ligand Protein / pharmacology
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Gene Expression Regulation, Neoplastic / drug effects
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Gene Expression Regulation, Neoplastic / genetics*
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Gene Expression Regulation, Neoplastic / radiation effects
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Humans
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Melanoma / genetics*
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Melanoma / metabolism*
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Oligonucleotide Array Sequence Analysis / methods*
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RNA, Small Interfering / genetics
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Signal Transduction / drug effects
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Signal Transduction / genetics
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Signal Transduction / radiation effects
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Ultraviolet Rays
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bcl-2 Homologous Antagonist-Killer Protein / genetics
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bcl-2 Homologous Antagonist-Killer Protein / metabolism
Substances
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Apoptosis Regulatory Proteins
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BAK1 protein, human
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DNA, Mitochondrial
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Fas Ligand Protein
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RNA, Small Interfering
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bcl-2 Homologous Antagonist-Killer Protein