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Use of a cDNA microarray to analyse gene expression patterns in human cancer
J DeRisi1, L Penland, P O Brown
1Howard Hughes Medical Institute, Department of Biochemistry, Stanford University Medical Center, California 94305, USA.
Abstract:
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.
Insights
Introducing a normal human chromosome 6 suppressed melanoma tumor growth and altered gene expression. This study identified specific gene expression changes linked to tumor suppression, offering new research avenues.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- Cancer development involves complex gene expression changes.
- Tumorigenicity in human melanoma cells (UACC-903) can be experimentally reversed.
- Introduction of normal human chromosome 6 suppresses melanoma growth and tumorigenicity.
Purpose of the Study:
- To identify gene expression differences associated with tumor suppression in melanoma cells.
- To investigate the genetic basis of the tumorigenic phenotype.
Main Methods:
- Utilized a high-density cDNA microarray with 1,161 DNA elements.
- Compared gene expression between UACC-903 melanoma cells and UACC-903 cells with introduced chromosome 6 (UACC-903(+6)).
- Employed fluorescently labeled probes for direct, internally controlled comparison of mRNA levels.
Main Results:
- Identified previously unrecognized alterations in specific gene expression patterns.
- Demonstrated that chromosome 6 introduction led to reduced growth rate, restored contact inhibition, and suppressed soft agar clonogenicity and in vivo tumorigenicity.
- Quantified relative mRNA abundance for each arrayed gene.
Conclusions:
- Specific gene expression alterations are linked to tumor suppression in melanoma.
- These findings provide new targets for investigating the genetic underpinnings of cancer.
- Further research into these identified genes can elucidate mechanisms of tumor suppression.