Genomic analysis of single cells from human basal cell cancer using laser-assisted capture microscopy.
In this study, we show that direct mutational analysis of genomic DNA can be performed on single somatic cells extracted from a frozen, immunohistochemically stained tissue section using laser-assisted capture microscopy. Eighty-nine single tumor cells were separately dissected from one case of human basal cell cancer (BCC) and p53 mutations were analyzed by direct semi-automated sequencing of PCR fragments. Amplification was obtained for at least one of the two analyzed exons from approximately 50% of the single tumor cells. Identical p53 mutations were found in widely spread areas of the tumor, suggesting a clonal proliferation originating from one cell. Interestingly, comparison between results of immunohistochemistry and genetic analysis of the single cells revealed the same p53 mutations irrespective of the p53 immunoreactivity. We propose that this approach has a great potential to allow investigation of genotypic differences in single cells and more specifically to resolve important and fundamental questions determining cancer heterogeneity.
Pontén
F
F
Department of Pathology, University Hospital, Uppsala, Sweden.
Williams
C
C
Ling
G
G
Ahmadian
A
A
Nistér
M
M
Lundeberg
J
J
Pontén
J
J
Uhlén
M
M
D000368:Aged; D000369:Aged, 80 and over; D002280:Basal Cell Carcinoma / Q000737:chemistry / Q000235:genetics* / Q000473:pathology; D002469:Cell Separation; D004252:DNA Mutational Analysis / Q000379:methods*; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D005091:Exons / Q000235:genetics; D005260:Female; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D007834:Lasers; D008853:Microscopy / Q000379:methods; D017354:Point Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D012878:Skin Neoplasms / Q000737:chemistry / Q000235:genetics*
D004273:DNA, Neoplasm