Quantitative real-time PCR identifies a critical region of deletion on 22q13 related to prognosis in oral cancer.
Quantitative real time PCR was performed on genomic DNA from 40 primary oral carcinomas and the normal adjacent tissues. The target genes ECGFB, DIA1, BIK, and PDGFB and the microsatellite markers D22S274 and D22S277, mapped on 22q13, were selected according to our previous loss of heterozygosity findings in head and neck tumors. Quantitative PCR relies on the comparison of the amount of product generated from a target gene and that generated from a disomic reference gene (GAPDH-housekeeping gene). Reactions have been performed with normal control in triplicates, using the 7700 Sequence Detection System (PE Applied Biosystems). Losses in the sequences D22S274 (22q13.31) and in the DIA1 (22q13.2-13.31) gene were detected in 10 out of 40 cases (25%) each. Statistically significant correlations were observed for patients with relative copy number loss of the marker D22S274 and stages T3-T4 of disease (P=0.025), family history of cancer (P=0.001), and death (P=0.021). Relative copy number loss involving the DIA1 gene was correlated to family history of cancer (P<0.001), death (P=0.002), and consumption of alcohol (P=0.026). Log-rank test revealed a significant decrease in survival (P=0.0018) for patients with DIA1 gene loss. Relative copy number losses detected in these sequences may be related to disease progression and a worse prognosis in patients with oral cancer.
Reis
Patricia P
PP
Department of Genetics, Universidade Estadual Paulista, Botucatu, Sâo Paulo, Brazil.
Rogatto
Silvia R
SR
Kowalski
Luiz P
LP
Nishimoto
Inês N
IN
Montovani
Jair C
JC
Corpus
George
G
Squire
Jeremy A
JA
Kamel-Reid
Suzanne
S
D000328:Adult; D000368:Aged; D000925:Anticoagulants; D017209:Apoptosis / Q000235:genetics; D051017:Apoptosis Regulatory Proteins; D000077214:Becaplermin; D002892:Chromosomes, Human, Pair 22 / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics* / Q000378:metabolism; D016228:Endothelial Growth Factors / Q000235:genetics; D005260:Female; D018628:Gene Dosage; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008565:Membrane Proteins; D018895:Microsatellite Repeats; D008875:Middle Aged; D024101:Mitochondrial Proteins; D009060:Mouth Floor / Q000473:pathology; D009062:Mouth Neoplasms / Q000235:genetics* / Q000473:pathology; D009363:Neoplasm Proteins / Q000235:genetics*; D010982:Platelet-Derived Growth Factor / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011379:Prognosis; D011506:Proteins / Q000235:genetics; D020574:Proto-Oncogene Proteins c-sis; D012680:Sensitivity and Specificity; D017384:Sequence Deletion; D014059:Tongue / Q000473:pathology; D014407:Tumor Cells, Cultured
D000925:Anticoagulants; D051017:Apoptosis Regulatory Proteins; C096820:BIK protein, human; D004273:DNA, Neoplasm; D016228:Endothelial Growth Factors; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D009363:Neoplasm Proteins; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D020574:Proto-Oncogene Proteins c-sis; D000077214:Becaplermin