Molecular genetic testing of uveal melanoma from routinely processed and stained cytology specimens.

Abstract

In the following study we investigated the utility of molecular genetic testing of the DNA extracted from routinely stained and processed smears from uveal melanoma (UM). Smears from five uveal melanoma cell lines and 12 primary tumors were prepared and stained with Papanicolaou and Romanowsky stains. Genotyping was carried out utilizing 14 microsatellite markers on chromosomes 3, 6 and 8. Mutational screening for alterations in GNAQ and GNA11 genes was carried out by restriction fragment length polymorphism. The results were compared to those obtained through direct sequencing of frozen tumor tissues. High quality DNA was extracted from the stained slides with no difference in the efficiency of DNA extraction between the two staining techniques. The extracted DNA was of adequate quality for genotyping and mutational screening. DNA extracted from approximately 200 tumor cells is sufficient for reproducible testing of allelic imbalances and for studying the common somatic mutations in GNAQ and GNA11 genes. In conclusion, we presented the feasibility of utilizing routinely stained cytology smears from UM for molecular genetic testing. The DNA obtained is of sufficient quality to carry out genotyping for markers on chromosome 3, 6 and 8, as well as screening for somatic mutations in GNAQ and GNA11 genes.

• Authors: Benjamin N Christopher, Colleen M Cebulla, Paul E Wakely Jr, Frederick H Davidorf, Mohamed H Abdel-Rahman
• Journal: Experimental eye research (Exp Eye Res) Vol. 93 Issue 5 Pg. 720-5 (Nov 2011) ISSN: 1096-0007 [Electronic] England
• PMID: 21945171 (Publication Type: Journal Article)
• Copyright: Copyright © 2011 Elsevier Ltd. All rights reserved.

Chemical References

• DNA, Neoplasm
• GNA11 protein, human
• GNAQ protein, human
• GTP-Binding Protein alpha Subunits
• GTP-Binding Protein alpha Subunits, Gq-G11

Topics

• Chromosomes, Human, Pair 3 (genetics)
• Chromosomes, Human, Pair 6 (genetics)
• Chromosomes, Human, Pair 8 (genetics)
• DNA Mutational Analysis
• DNA, Neoplasm (analysis, isolation & purification)
• Feasibility Studies
• GTP-Binding Protein alpha Subunits (genetics)
• GTP-Binding Protein alpha Subunits, Gq-G11
• Genotype
• Genotyping Techniques
• Histocytological Preparation Techniques
• Humans
• Melanoma (genetics, pathology)
• Microsatellite Repeats (genetics)
• Molecular Diagnostic Techniques
• Polymorphism, Restriction Fragment Length
• Tumor Cells, Cultured
• Uveal Neoplasms (genetics, pathology)