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P53 deletions/mutations in Brazilian adult T-cell leukemia patients


, : P53 deletions/mutations in Brazilian adult T-cell leukemia patients. Blood 96(11 Part 2): 229b, November 16

Background: P53 is a regulator of cell cycling. Mutations or deletions in this gene are associated with malignancies. Adult T-cell leukemia (ATL) is one the diseases associated with HTLV-1 infection. The mechanisms of HTLV-I mediated cellular transformation are not clearly understood. However, events such as viral deletions with loss of down regulation of viral transcription activators, and p53 and/or p16 genetic alterations have been implicated in ATL pathogenesis. We evaluated viral deletions and p53 alterations in a cohort of ATL patients from Brazil in search of correlates of ATL development. Methods: We looked for HTLV-1 proviral sequences in specimens from 32 ATL patients using long PCR. We also searched for alterations in exons 4, 5, 6, 7, and 8 of p53 using PCR followed by SSCP analysis. Amplified bands of short size in products of long PCR for HTLV-I suggest deletions of proviral sequences, while alterations in the mobility of single-stranded DNA amplified products indicate mutation of p53 sequences. Therefore, samples with altered mobility in the SSCP assay were subjected to direct sequencing of PCR products. The Duffy blood group gene was co-amplified as control. Results: We found that 9/32 (28%) samples had defective HTLV-I provirus of type 1, and 3 (10%) had defective HTLV-1 provirus type 2, for a total of 12/32 ATL patients with altered p53 sequences. In addition, we could not amplify p53 genes in 10/26 (38%) of the samples, leading us to suspect that deletions and gross alterations of p53 sequences prevented primer annealing and amplification. Two of the 26 (7%) samples had p53 mutations that are known to alter gene function/expression. In one there was a T>C mutation in exon 4 predicted to induce a D42N mutation. In the other we found a G>A mutation in exon 4 predicted to generate a premature stop codon at residue 91 (W91 stop). A combination of HTLV-1 defective provirus and p53 deletion was found in 3 cases. Conclusion: HTLV-I proviral sequence deletions and p53 deletions can be detected in peripheral blood cells of ATL patients and confirms the contribution of these alterations to the pathogenesis of ATL.

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