Differential gene expression profiling of primary cutaneous melanoma and sentinel lymph node metastases
Limited understanding of molecular mechanisms of metastasis in melanoma contributes to the absence of effective treatments. Increased knowledge of alterations in genes that underpin critical molecular events that lead to metastasis is essential. We have investigated the gene expression profiles of p...
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| Veröffentlicht in: | Modern pathology 2012-06, Vol.25 (6), p.828-837 |
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| creator | Koh, Stephen S Wei, Jia-Perng J Li, Xinmin Huang, Rong R Doan, Ngan B Scolyer, Richard A Cochran, Alistair J Binder, Scott W |
| description | Limited understanding of molecular mechanisms of metastasis in melanoma contributes to the absence of effective treatments. Increased knowledge of alterations in genes that underpin critical molecular events that lead to metastasis is essential. We have investigated the gene expression profiles of primary melanomas and melanoma metastases in sentinel lymph nodes. A total of 19 samples (10 primary melanomas and 9 sentinel lymph node metastases) were evaluated. Melanoma cells were dissected from tissue blocks. Total mRNA was isolated, amplified, and labeled using an Ambion Recover All Total Nucleic Acid Isolation kit, Nu-GEN WT-Ovation formalin-fixed, paraffin-embedded RNA Amplification System, and FL-Ovation cDNA Biotin Module V2, respectively. Samples were hybridized to the Affymetrix Gene Chip Human U133 Plus 2.0 Array. Data were analyzed using Partek Genomics Suite Version 6.4. Genes selected showed ≥2-fold difference in expression and P |
| doi_str_mv | 10.1038/modpathol.2012.32 |
| format | Article |
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Increased knowledge of alterations in genes that underpin critical molecular events that lead to metastasis is essential. We have investigated the gene expression profiles of primary melanomas and melanoma metastases in sentinel lymph nodes. A total of 19 samples (10 primary melanomas and 9 sentinel lymph node metastases) were evaluated. Melanoma cells were dissected from tissue blocks. Total mRNA was isolated, amplified, and labeled using an Ambion Recover All Total Nucleic Acid Isolation kit, Nu-GEN WT-Ovation formalin-fixed, paraffin-embedded RNA Amplification System, and FL-Ovation cDNA Biotin Module V2, respectively. Samples were hybridized to the Affymetrix Gene Chip Human U133 Plus 2.0 Array. Data were analyzed using Partek Genomics Suite Version 6.4. Genes selected showed ≥2-fold difference in expression and P<5.00E−2. Validation studies used standard immunohistochemical assays. Hierarchical clustering disclosed two distinct groups: 10 primary melanomas and 9 sentinel lymph node metastases. Gene expression analysis identified 576 genes that showed significant differential expression. Most differences reflected decreased gene expression in metastases relative to primaries. Reduced gene expression in primaries was less frequent and less dramatic. Genes significantly increased or decreased in sentinel lymph node metastases were active in cell adhesion/structural integrity, tumor suppression, cell cycle regulation, and apoptosis. Validation studies indicate that MAGEC1 (melanoma antigen family C1) and FCRL1 (Fc receptor-like 1) are involved in melanoma progression. There are striking differential gene expression patterns between primary and nodally metastatic melanomas. Similar findings were seen with autologous paired primary melanomas and sentinel lymph node metastases, supporting involvement of these gene alterations in evolution of metastases. With further study, it may be possible to determine the exact sequence of molecular events that underlie melanoma metastases.</description><identifier>ISSN: 0893-3952</identifier><identifier>EISSN: 1530-0285</identifier><identifier>DOI: 10.1038/modpathol.2012.32</identifier><identifier>PMID: 22411186</identifier><identifier>CODEN: MODPEO</identifier><language>eng</language><publisher>New York: Elsevier Inc</publisher><subject>631/1647/2217/2018 ; 692/699/67/1813/1634 ; 692/699/67/322 ; 692/700/139/422 ; Adult ; Aged ; Aged, 80 and over ; Antigens, Neoplasm - genetics ; Biomarkers ; Biomarkers, Tumor - analysis ; Biomarkers, Tumor - genetics ; Cancer ; Cell cycle ; Cluster Analysis ; DNA microarray ; Female ; formalin-fixed, paraffin-embedded tissue ; Gene expression ; Gene Expression Profiling - methods ; Gene Expression Regulation, Neoplastic ; Genetic Predisposition to Disease ; Genomes ; Genomics ; Humans ; Immunohistochemistry ; Laboratory Medicine ; Los Angeles ; Lymph Nodes - chemistry ; Lymph Nodes - pathology ; Lymphatic Metastasis ; Lymphatic system ; Male ; Medicine ; Medicine & Public Health ; Melanoma ; Melanoma - chemistry ; Melanoma - genetics ; Melanoma - secondary ; Membrane Proteins - genetics ; Metastasis ; Middle Aged ; Neoplasm Proteins - genetics ; New South Wales ; Oligonucleotide Array Sequence Analysis ; Oncology ; original-article ; Paraffin Embedding ; Pathology ; Phenotype ; Principal Component Analysis ; Reproducibility of Results ; RNA, Messenger - analysis ; sentinel lymph node ; Sentinel Lymph Node Biopsy ; Skin Neoplasms - chemistry ; Skin Neoplasms - genetics ; Skin Neoplasms - pathology ; Validation studies</subject><ispartof>Modern pathology, 2012-06, Vol.25 (6), p.828-837</ispartof><rights>2012 United States & Canadian Academy of Pathology</rights><rights>United States and Canadian Academy of Pathology, Inc. 2012</rights><rights>Copyright Nature Publishing Group Jun 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c566t-33494b90b58f97d3f6dd99837a19b4fe4f6373757a1a6399732afccf391d3dd43</citedby><cites>FETCH-LOGICAL-c566t-33494b90b58f97d3f6dd99837a19b4fe4f6373757a1a6399732afccf391d3dd43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22411186$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Koh, Stephen S</creatorcontrib><creatorcontrib>Wei, Jia-Perng J</creatorcontrib><creatorcontrib>Li, Xinmin</creatorcontrib><creatorcontrib>Huang, Rong R</creatorcontrib><creatorcontrib>Doan, Ngan B</creatorcontrib><creatorcontrib>Scolyer, Richard A</creatorcontrib><creatorcontrib>Cochran, Alistair J</creatorcontrib><creatorcontrib>Binder, Scott W</creatorcontrib><title>Differential gene expression profiling of primary cutaneous melanoma and sentinel lymph node metastases</title><title>Modern pathology</title><addtitle>Mod Pathol</addtitle><addtitle>Mod Pathol</addtitle><description>Limited understanding of molecular mechanisms of metastasis in melanoma contributes to the absence of effective treatments. Increased knowledge of alterations in genes that underpin critical molecular events that lead to metastasis is essential. We have investigated the gene expression profiles of primary melanomas and melanoma metastases in sentinel lymph nodes. A total of 19 samples (10 primary melanomas and 9 sentinel lymph node metastases) were evaluated. Melanoma cells were dissected from tissue blocks. Total mRNA was isolated, amplified, and labeled using an Ambion Recover All Total Nucleic Acid Isolation kit, Nu-GEN WT-Ovation formalin-fixed, paraffin-embedded RNA Amplification System, and FL-Ovation cDNA Biotin Module V2, respectively. Samples were hybridized to the Affymetrix Gene Chip Human U133 Plus 2.0 Array. Data were analyzed using Partek Genomics Suite Version 6.4. Genes selected showed ≥2-fold difference in expression and P<5.00E−2. Validation studies used standard immunohistochemical assays. Hierarchical clustering disclosed two distinct groups: 10 primary melanomas and 9 sentinel lymph node metastases. Gene expression analysis identified 576 genes that showed significant differential expression. Most differences reflected decreased gene expression in metastases relative to primaries. Reduced gene expression in primaries was less frequent and less dramatic. Genes significantly increased or decreased in sentinel lymph node metastases were active in cell adhesion/structural integrity, tumor suppression, cell cycle regulation, and apoptosis. Validation studies indicate that MAGEC1 (melanoma antigen family C1) and FCRL1 (Fc receptor-like 1) are involved in melanoma progression. There are striking differential gene expression patterns between primary and nodally metastatic melanomas. Similar findings were seen with autologous paired primary melanomas and sentinel lymph node metastases, supporting involvement of these gene alterations in evolution of metastases. 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chemistry</subject><subject>Lymph Nodes - pathology</subject><subject>Lymphatic Metastasis</subject><subject>Lymphatic system</subject><subject>Male</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Melanoma</subject><subject>Melanoma - chemistry</subject><subject>Melanoma - genetics</subject><subject>Melanoma - secondary</subject><subject>Membrane Proteins - genetics</subject><subject>Metastasis</subject><subject>Middle Aged</subject><subject>Neoplasm Proteins - genetics</subject><subject>New South Wales</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Oncology</subject><subject>original-article</subject><subject>Paraffin Embedding</subject><subject>Pathology</subject><subject>Phenotype</subject><subject>Principal Component Analysis</subject><subject>Reproducibility of Results</subject><subject>RNA, Messenger - analysis</subject><subject>sentinel lymph node</subject><subject>Sentinel Lymph Node Biopsy</subject><subject>Skin Neoplasms - 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Increased knowledge of alterations in genes that underpin critical molecular events that lead to metastasis is essential. We have investigated the gene expression profiles of primary melanomas and melanoma metastases in sentinel lymph nodes. A total of 19 samples (10 primary melanomas and 9 sentinel lymph node metastases) were evaluated. Melanoma cells were dissected from tissue blocks. Total mRNA was isolated, amplified, and labeled using an Ambion Recover All Total Nucleic Acid Isolation kit, Nu-GEN WT-Ovation formalin-fixed, paraffin-embedded RNA Amplification System, and FL-Ovation cDNA Biotin Module V2, respectively. Samples were hybridized to the Affymetrix Gene Chip Human U133 Plus 2.0 Array. Data were analyzed using Partek Genomics Suite Version 6.4. Genes selected showed ≥2-fold difference in expression and P<5.00E−2. Validation studies used standard immunohistochemical assays. Hierarchical clustering disclosed two distinct groups: 10 primary melanomas and 9 sentinel lymph node metastases. Gene expression analysis identified 576 genes that showed significant differential expression. Most differences reflected decreased gene expression in metastases relative to primaries. Reduced gene expression in primaries was less frequent and less dramatic. Genes significantly increased or decreased in sentinel lymph node metastases were active in cell adhesion/structural integrity, tumor suppression, cell cycle regulation, and apoptosis. Validation studies indicate that MAGEC1 (melanoma antigen family C1) and FCRL1 (Fc receptor-like 1) are involved in melanoma progression. There are striking differential gene expression patterns between primary and nodally metastatic melanomas. Similar findings were seen with autologous paired primary melanomas and sentinel lymph node metastases, supporting involvement of these gene alterations in evolution of metastases. 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| subjects | 631/1647/2217/2018 692/699/67/1813/1634 692/699/67/322 692/700/139/422 Adult Aged Aged, 80 and over Antigens, Neoplasm - genetics Biomarkers Biomarkers, Tumor - analysis Biomarkers, Tumor - genetics Cancer Cell cycle Cluster Analysis DNA microarray Female formalin-fixed, paraffin-embedded tissue Gene expression Gene Expression Profiling - methods Gene Expression Regulation, Neoplastic Genetic Predisposition to Disease Genomes Genomics Humans Immunohistochemistry Laboratory Medicine Los Angeles Lymph Nodes - chemistry Lymph Nodes - pathology Lymphatic Metastasis Lymphatic system Male Medicine Medicine & Public Health Melanoma Melanoma - chemistry Melanoma - genetics Melanoma - secondary Membrane Proteins - genetics Metastasis Middle Aged Neoplasm Proteins - genetics New South Wales Oligonucleotide Array Sequence Analysis Oncology original-article Paraffin Embedding Pathology Phenotype Principal Component Analysis Reproducibility of Results RNA, Messenger - analysis sentinel lymph node Sentinel Lymph Node Biopsy Skin Neoplasms - chemistry Skin Neoplasms - genetics Skin Neoplasms - pathology Validation studies |
| title | Differential gene expression profiling of primary cutaneous melanoma and sentinel lymph node metastases |
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