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Differential genetic expression in large versus small clear cell renal cell carcinoma: results from microarray analysis.

Bratslavsky G, Sanford T, Srinivasan R, Aprelikova O, Liu J, Quezado M, Merino M, Linehan WM - J Cancer (2011)

Bottom Line: P<0.01 was considered statistically significant.Paired t-test showed 860 differentially expressed genes in the T1b vs T1a group, a number far greater than expected due to chance alone.When analyzed by gene function, most differences were observed in genes involved in DNA replication and in cytokine signaling.

View Article: PubMed Central - PubMed

Affiliation: 1. Urologic Oncology Branch, National Cancer Institute, Bethesda, MD, USA.

ABSTRACT

Purpose: Tumor growth and progression requires multiple steps and genetic alterations. The molecular events that occur as tumors increase in size are unknown. Patients with von Hippel-Lindau (VHL) provide a unique opportunity to study molecular alterations during tumor growth as these patients develop multiple bilateral renal tumors. To better characterize biologic events associated with tumor growth, we evaluated the alterations in gene expression in large versus small renal tumors removed from the same kidney of the same individuals.

Experimental design: We reviewed pathology reports from patients who underwent partial nephrectomies at the National Cancer Institute for multiple tumors. We identified 11 patients who fulfilled the following inclusion criteria: 1) The patient must have had a surgical resection of more than one solid tumor from the same kidney during the same operation; 2) Among the solid tumors at least one must have been greater than 3 cm in the largest dimension and at least one less than 2 cm; 3) the nuclear Furhman grade for both larger and smaller solid tumors was identical; 4) a portion of each tumor was procured and snap frozen after surgical removal; 5) Hematoxylin and eosin staining of the frozen sample confirmed clear cell carcinoma to be present in at least 80% of the section.Affymetrix platform and protocol for gene expression arrays were used. RNA from the frozen large and small tumor samples was extracted using Trizol-Chlorophorm method. The RNA was then reverse transcribed, labeled, fragmented, and hybridized on to an Affymetrix U133 Plus 2.0 array that contains 54,000 probe sets representing 24,568 genes. Analysis included unsupervised clustering and chromosomal analysis. The paired t-test was performed to compare gene expression levels in small and large tumors. P<0.01 was considered statistically significant.

Results: Gene expression profiles were assessed for 22 tumors (11 patients). Upon unsupervised clustering the pairs with larger tumor volume difference clustered separately from pairs with smaller volume difference. Chromosomal analysis revealed few consistent changes other than reduced expression of chromosome 3p25 among all tumors. Paired t-test showed 860 differentially expressed genes in the T1b vs T1a group, a number far greater than expected due to chance alone. When analyzed by gene function, most differences were observed in genes involved in DNA replication and in cytokine signaling.

Conclusions: This study demonstrates that as tumors increase in size there is an increasing difference in gene expression. Unsupervised clustering analysis confirms that as the volume difference increases there are a distinct set of genes that are regulated either as a response to a tumor's growth or as an early event that causes the tumor to grow. While we did not observe chromosomal instability, we did note differences in expression of individual transcripts as tumors grew larger.

No MeSH data available.


Related in: MedlinePlus

Comparative genomic microarray analysis - global chromosomal heat map. No consistent differences in expression on chromosomal level between large and small tumors were observed. Note that the changes between individuals are greater than changes between tumors of different size in a given individual.
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Figure 5: Comparative genomic microarray analysis - global chromosomal heat map. No consistent differences in expression on chromosomal level between large and small tumors were observed. Note that the changes between individuals are greater than changes between tumors of different size in a given individual.

Mentions: The first part of the cGMA chromosomal analysis was an examination of the expression pattern for chromosome 3. Since all the patients had a confirmed VHL mutation, as expected the level of expression was decreased in chromosome 3p25 in all samples when compared to normal renal parenchyma (Figure 4). Interestingly, there is was also a decrease in gene expression near chromosome 3p21. Full chromosomal analysis revealed that the expression profiles within each pair appeared similar. No consistent differences in expression on chromosomal level between large and small tumors were observed. (Figure 5).


Differential genetic expression in large versus small clear cell renal cell carcinoma: results from microarray analysis.

Bratslavsky G, Sanford T, Srinivasan R, Aprelikova O, Liu J, Quezado M, Merino M, Linehan WM - J Cancer (2011)

Comparative genomic microarray analysis - global chromosomal heat map. No consistent differences in expression on chromosomal level between large and small tumors were observed. Note that the changes between individuals are greater than changes between tumors of different size in a given individual.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3100679&req=5

Figure 5: Comparative genomic microarray analysis - global chromosomal heat map. No consistent differences in expression on chromosomal level between large and small tumors were observed. Note that the changes between individuals are greater than changes between tumors of different size in a given individual.
Mentions: The first part of the cGMA chromosomal analysis was an examination of the expression pattern for chromosome 3. Since all the patients had a confirmed VHL mutation, as expected the level of expression was decreased in chromosome 3p25 in all samples when compared to normal renal parenchyma (Figure 4). Interestingly, there is was also a decrease in gene expression near chromosome 3p21. Full chromosomal analysis revealed that the expression profiles within each pair appeared similar. No consistent differences in expression on chromosomal level between large and small tumors were observed. (Figure 5).

Bottom Line: P<0.01 was considered statistically significant.Paired t-test showed 860 differentially expressed genes in the T1b vs T1a group, a number far greater than expected due to chance alone.When analyzed by gene function, most differences were observed in genes involved in DNA replication and in cytokine signaling.

View Article: PubMed Central - PubMed

Affiliation: 1. Urologic Oncology Branch, National Cancer Institute, Bethesda, MD, USA.

ABSTRACT

Purpose: Tumor growth and progression requires multiple steps and genetic alterations. The molecular events that occur as tumors increase in size are unknown. Patients with von Hippel-Lindau (VHL) provide a unique opportunity to study molecular alterations during tumor growth as these patients develop multiple bilateral renal tumors. To better characterize biologic events associated with tumor growth, we evaluated the alterations in gene expression in large versus small renal tumors removed from the same kidney of the same individuals.

Experimental design: We reviewed pathology reports from patients who underwent partial nephrectomies at the National Cancer Institute for multiple tumors. We identified 11 patients who fulfilled the following inclusion criteria: 1) The patient must have had a surgical resection of more than one solid tumor from the same kidney during the same operation; 2) Among the solid tumors at least one must have been greater than 3 cm in the largest dimension and at least one less than 2 cm; 3) the nuclear Furhman grade for both larger and smaller solid tumors was identical; 4) a portion of each tumor was procured and snap frozen after surgical removal; 5) Hematoxylin and eosin staining of the frozen sample confirmed clear cell carcinoma to be present in at least 80% of the section.Affymetrix platform and protocol for gene expression arrays were used. RNA from the frozen large and small tumor samples was extracted using Trizol-Chlorophorm method. The RNA was then reverse transcribed, labeled, fragmented, and hybridized on to an Affymetrix U133 Plus 2.0 array that contains 54,000 probe sets representing 24,568 genes. Analysis included unsupervised clustering and chromosomal analysis. The paired t-test was performed to compare gene expression levels in small and large tumors. P<0.01 was considered statistically significant.

Results: Gene expression profiles were assessed for 22 tumors (11 patients). Upon unsupervised clustering the pairs with larger tumor volume difference clustered separately from pairs with smaller volume difference. Chromosomal analysis revealed few consistent changes other than reduced expression of chromosome 3p25 among all tumors. Paired t-test showed 860 differentially expressed genes in the T1b vs T1a group, a number far greater than expected due to chance alone. When analyzed by gene function, most differences were observed in genes involved in DNA replication and in cytokine signaling.

Conclusions: This study demonstrates that as tumors increase in size there is an increasing difference in gene expression. Unsupervised clustering analysis confirms that as the volume difference increases there are a distinct set of genes that are regulated either as a response to a tumor's growth or as an early event that causes the tumor to grow. While we did not observe chromosomal instability, we did note differences in expression of individual transcripts as tumors grew larger.

No MeSH data available.


Related in: MedlinePlus