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TSG101 is not mutated in lung cancer but a shortened transcript is frequently expressed in small cell lung cancer
1Department of Thoracic/Head and Neck Medical Oncology, The University of Texas-Houston, M. D. Anderson Cancer Center, 77030, USA.
Abstract:
TSG101 is a candidate tumor suppressor gene whose deletion in NIH3T3 cells leads to spontaneous lung metastases in nude mice. Aberrant transcripts of TSG101 have been identified in 47% of primary breast carcinomas, without evidence of intragenic deletions at the TSG101 locus on 11p15. To investigate the possible role of TSG101 in lung cancer, which often shows 11p allele loss, we performed transcript analysis and mutational analysis of TSG101 in lung cancer cell lines. Reverse transcriptase RT-PCR and Northern analysis detected a common TSG101 transcript, shortened because of an internal deletion, which was expressed simultaneously with the wild-type transcript in 89% of small cell lung cancer (SCLC) lines. In contrast, the wild-type transcript was expressed alone in normal tissues, primary non-small cell lung cancer (NSCLC) specimens, and the majority of NSCLC cell lines. Sequence of the shortened SCLC transcript was identical to that of the most common aberrant transcript identified in breast cancer, consisting of a deletion of exons 2-4 and part of 1 and 5. Southern analysis of SCLC lines expressing the shortened transcript did not detect any intragenic deletions. Single strand conformational polymorphism (SSCP) analysis and direct sequencing of TSG101 cDNAs also identified no mutations or deletions. These results suggest that TSG101 is not mutated in lung cancer but that aberrant splicing of TSG101 occurs in SCLC.
Insights
The tumor suppressor gene TSG101, when aberrantly spliced, is found in most small cell lung cancer (SCLC) cell lines. This aberrant splicing, not mutation, is linked to SCLC development.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Genetics
Background:
- TSG101 is a candidate tumor suppressor gene.
- Aberrant TSG101 transcripts are found in breast cancer.
- Lung cancer often exhibits 11p allele loss, the locus of TSG101.
Purpose of the Study:
- Investigate the role of TSG101 in lung cancer.
- Analyze TSG101 transcripts and mutations in lung cancer cell lines.
Main Methods:
- Reverse transcriptase RT-PCR and Northern analysis for transcript detection.
- Southern analysis for intragenic deletions.
- Single strand conformational polymorphism (SSCP) analysis and direct sequencing for mutations.
Main Results:
- A shortened TSG101 transcript due to internal deletion was found in 89% of small cell lung cancer (SCLC) lines, co-expressed with the wild-type transcript.
- Normal tissues and non-small cell lung cancer (NSCLC) predominantly expressed only the wild-type transcript.
- No intragenic deletions or mutations in TSG101 were detected in SCLC lines with the shortened transcript.
Conclusions:
- TSG101 is not mutated in lung cancer.
- Aberrant splicing of TSG101, leading to a shortened transcript, occurs in SCLC.
- This aberrant splicing may play a role in SCLC development.