Role of TSG101 in uterine cervix cancer

Objective. TSG101 was first described as a possible tumor suppressor gene i n breast cancer. To determine whether TSG101 might play a role in cervical carcinogenesis, we examined a panel of cervical cancer cell lines and prima ry tumor specimens for transcript abnormalities and mutations in TSG101, Methods. Total RNA was derived from cell line cultures or primary tumor spe cimens. We performed nested reverse transcription polymerase chain reaction (RT-PCR) with eight overlapping primer sets, followed by single-strand con formational polymorphism (SSCP) analysis, to screen for mutations in the TS G101 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 cervica l tumors and 8 cervical carcinoma cell lines, Two polymorphisms were identi fied, neither of which resulted in an altered amino acid sequence. Transcri pt analysis was performed on a subset of 16 primary cervix tumors and 6 cer vix carcinoma cell lines, The wild-type transcript (1228 bp) was the domina nt transcript expressed in all samples, A transcript measuring 330 bp was d etected in 5 of 6 cell lines and 11 of 16 primary tumor specimens. Conclusion. Our results suggest that mutations in TSG101 rarely occur in ca rcinomas of the uterine cervix, However, the presence of minor aberrant TSG 101 transcripts is a common feature. The relationship between aberrant tran scription and carcinogenesis should be further investigated. (C) 1999 Acade mic Press.