Absence of TSG101 transcript abnormalities in human cancers.
The human TSG101 gene was cloned and mapped to chromosome 11p15, a site suspected to contain a tumor suppressor gene involved in a variety of human cancers. Subsequent investigation described the presence of abnormally spliced transcripts and point mutations of TSG101 in breast cancer. Thus, we performed RT-PCR amplification of the entire open reading frame of TSG101 to test for aberrant transcripts in various human tumor cell lines derived from breast, bladder, head and neck, and lung cancer. In addition, we performed RT-PCR on cDNA from primary human breast and Wilms' tumor tissue. We found a single band of the expected size in 10 of 11 breast cancers and 6 of 6 Wilms' tumor samples after the first round of PCR. The remaining breast cancer sample displayed a barely visible smaller band. However, aberrant bands appeared in most cases after performing nested PCR casting doubt on the physiologic relevance of these spliced variants. We then searched for small intragenic mutations by complete sequence analysis of TSG101 in breast cancer cell lines and tumors, as well as in Wilms' tumors and normal fetal and adult kidney. No point mutations were found in any of the samples, including four breast tumors with chromosomal loss at 11p15. We found no consistent evidence of aberrant splicing or point mutations in breast cancer or Wilms' tumor suggesting that TSG101 is not a primary target of inactivation in human cancer.
Trink
B
B
Department of Otolarynogology-Head and Neck Surgery, The Johns Hopkins University, Baltimore, Maryland 21205-2196, USA.
Pai
S I
SI
Spunt
S L
SL
Raman
V
V
Cairns
P
P
Jen
J
J
Gabrielson
E
E
Sukumar
S
S
Sidransky
D
D
D000328:Adult; D001943:Breast Neoplasms / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D056827:Endosomal Sorting Complexes Required for Transport; D005260:Female; D006801:Humans; D009369:Neoplasms / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D012334:RNA, Neoplasm / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D014407:Tumor Cells, Cultured; D009396:Wilms Tumor / Q000235:genetics*; D000099177:Tsg101 Protein
D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D012334:RNA, Neoplasm; D014157:Transcription Factors; D000099177:Tsg101 Protein