DeRisi J, Penland L, Brown P O, Bittner M L, Meltzer P S, Ray M, Chen Y, Su Y A, Trent J M
Howard Hughes Medical Institute, Department of Biochemistry, Stanford University Medical Center, California 94305, USA.
Nat Genet. 1996 Dec;14(4):457-60. doi: 10.1038/ng1296-457.
The development and progression of cancer and the experimental reversal of tumorigenicity are accompanied by complex changes in patterns of gene expression. Microarrays of cDNA provide a powerful tool for studying these complex phenomena. The tumorigenic properties of a human melanoma cell line, UACC-903, can be suppressed by introduction of a normal human chromosome 6, resulting in a reduction of growth rate, restoration of contact inhibition, and suppression of both soft agar clonogenicity and tumorigenicity in nude mice. We used a high density microarray of 1,161 DNA elements to search for differences in gene expression associated with tumour suppression in this system. Fluorescent probes for hybridization were derived from two sources of cellular mRNA [UACC-903 and UACC-903(+6)] which were labelled with different fluors to provide a direct and internally controlled comparison of the mRNA levels corresponding to each arrayed gene. The fluorescence signals representing hybridization to each arrayed gene were analysed to determine the relative abundance in the two samples of mRNAs corresponding to each gene. Previously unrecognized alterations in the expression of specific genes provide leads for further investigation of the genetic basis of the tumorigenic phenotype of these cells.
癌症的发展与进程以及肿瘤发生性的实验性逆转都伴随着基因表达模式的复杂变化。cDNA微阵列提供了一个研究这些复杂现象的有力工具。通过导入一条正常人类6号染色体，可以抑制人黑色素瘤细胞系UACC - 903的致瘤特性，从而导致生长速率降低、接触抑制恢复以及软琼脂克隆形成能力和裸鼠致瘤性均受到抑制。我们使用了一个包含1161个DNA元件的高密度微阵列来寻找与该系统中肿瘤抑制相关的基因表达差异。用于杂交的荧光探针来源于两种细胞mRNA [UACC - 903和UACC - 903(+6)]，它们用不同的荧光染料标记，以便对与每个阵列基因相对应的mRNA水平进行直接且内部对照的比较。分析代表与每个阵列基因杂交的荧光信号，以确定与每个基因相对应的两种mRNA样品中的相对丰度。特定基因表达中先前未被识别的改变为进一步研究这些细胞致瘤表型的遗传基础提供了线索。
