Zhang Qiuyang, Wu Jun, Nguyen Anhthu, Wang Bi-Dar, He Ping, Laurent Georges St, Rennert Owen M, Su Yan A
Department of Biochemistry and Molecular Biology and the Catherine Birch McCormick Genomics Center, The George Washington University School of Medicine and Health Sciences, Ross Hall, Room 555, 2300 I Street NW, Washington, DC 20037, USA.
Apoptosis. 2008 Aug;13(8):993-1004. doi: 10.1007/s10495-008-0231-8.
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.
人恶性黑色素瘤细胞系UACC903对凋亡具有抗性，而6号染色体介导的抑制细胞系UACC903(+6)则对凋亡敏感。在此，我们描述了这种差异背后不同分子途径的鉴定。使用我们最近开发的聚焦线粒体的cDNA微阵列，我们鉴定出154个差异表达基因，包括促凋亡基因（BAK1 [6p21.3]、BCAP31、BNIP1、CASP3、CASP6、FAS、FDX1、FDXR、TNFSF10和VDAC1）和抗凋亡基因（BCL2L1、CLN3和MCL1）。在紫外线处理前，这些促凋亡和抗凋亡基因在UACC903(+6)中的表达高于UACC903，且在紫外线处理后发生了改变。qRT-PCR和蛋白质印迹验证了微阵列结果。我们的生物信息学分析将这些基因映射到分别预测UACC903和UACC903(+6)对凋亡抗性和敏感性的不同分子途径。通过FAS配体诱导的细胞死亡和BAK1蛋白的siRNA敲低在功能上证实了这些途径。这些结果证明了UACC903和UACC903(+6)细胞系存活和凋亡背后的不同分子途径。
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