Truncated TSG101 transcripts in human leukemia and lymphoma cell lines

Abstract

Inactivation of the TSG101 gene has been shown to induce cellular transformation of NIH3T3 fibroblasts, and aberrant TSG101 transcripts have been observed not only in various human solid tumors but also in hematopoietic malignant disorders. In the present study, we performed nested reverse transcription/polymerase chain reaction (RT-PCR) analysis to identify aberrant TSG101 transcripts in 43 human leukemia and lymphoma cell lines. We could detect only a single band of the wild-type transcript with the expected size in virtually all cell lines after the first round of PCR. As in the case with various human solid tumors, the smaller TSG101 transcripts appeared in most of these cell lines after the second round of PCR. Thus, the expression level of the variant transcripts was extremely low as compared with that of the wild-type transcript, and this finding was also confirmed by Northern blot analysis. Identification of various truncated transcripts with extensive deletions in the TSG101 coding region was confirmed by means of sequencing analysis, and expression of these transcripts did not appear to be associated with a specific type of hematopoietic malignant disorder. Southern blot analysis did not indicate any gross TSG101 gene rearrangement. The truncated transcripts were also detected in normal peripheral blood leukocytes. Our results suggest that the truncated TSG101 transcripts are definitely detectable in various human leukemia and lymphoma cell lines, but do not support the notion that the variant transcripts may have a major functional relevance in the pathogenesis of human hematopoietic malignant disorder.