Role of TSG101 in uterine cervix cancer.
OBJECTIVE
TSG101 was first described as a possible tumor suppressor gene in breast cancer. To determine whether TSG101 might play a role in cervical carcinogenesis, we examined a panel of cervical cancer cell lines and primary tumor specimens for transcript abnormalities and mutations in TSG101.


METHODS
Total RNA was derived from cell line cultures or primary tumor specimens. We performed nested reverse transcription polymerase chain reaction (RT-PCR) with eight overlapping primer sets, followed by single-strand conformational polymorphism (SSCP) analysis, to screen for mutations in the TSG101 open reading frame. Representative normal and shifted SSCP bands were sequenced. To identify abnormal-sized transcripts, we performed RT-PCR with primers flanking the open reading frame followed by gel electrophoresis.


RESULTS
Mutational analysis was performed on cDNAs from 20 primary cervical tumors and 8 cervical carcinoma cell lines. Two polymorphisms were identified, neither of which resulted in an altered amino acid sequence. Transcript analysis was performed on a subset of 16 primary cervix tumors and 6 cervix carcinoma cell lines. The wild-type transcript (1228 bp) was the dominant transcript expressed in all samples. A transcript measuring 330 bp was detected in 5 of 6 cell lines and 11 of 16 primary tumor specimens.


CONCLUSIONS
Our results suggest that mutations in TSG101 rarely occur in carcinomas of the uterine cervix. However, the presence of minor aberrant TSG101 transcripts is a common feature. The relationship between aberrant transcription and carcinogenesis should be further investigated.
O'Boyle
J D
JD
Department of Obstetrics and Gynecology, The Nancy B. and Jake L. Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75235, USA.
Proctor
M L
ML
Fong
K M
KM
Lin
W M
WM
Miller
D S
DS
Muller
C Y
CY
D017766:Acid Anhydride Hydrolases; D002874:Chromosome Mapping; D002880:Chromosomes, Human, Pair 11; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D056827:Endosomal Sorting Complexes Required for Transport; D005260:Female; D016147:Genes, Tumor Suppressor; D006801:Humans; D009154:Mutation; D009363:Neoplasm Proteins; D016366:Open Reading Frames; D011506:Proteins / Q000235:genetics; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000209:etiology*; D000099177:Tsg101 Protein; D000099170:Bis(5'-adenosyl)triphosphatase
D017766:Acid Anhydride Hydrolases; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D009363:Neoplasm Proteins; D011506:Proteins; D014157:Transcription Factors; D000099177:Tsg101 Protein; D000099170:Bis(5'-adenosyl)triphosphatase