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Updated: Sep 10, 2026
Comparative Lesions Analysis Through a Targeted Sequencing Approach
Published on: November 5, 2019
Analysis of Fas gene mutations on laser capture microdissected specimens from renal cell carcinoma
Hitoshi Takayama1, Tetsuya Takakuwa, Yuichi Tsujimoto
1Department of Pathology, Osaka University Medical School, Suita, Osaka 565-0871, Japan.
Abstract:
Renal cell carcinoma (RCC) expresses Fas antigen on the cell surface, and thus could be sensitive to apoptosis induced by the binding of Fas ligand. Fas gene mutations might be involved in the development of RCC. Fas gene mutations were examined in genomic DNA extracted from RCC lesions. With use of laser capture methods, one RCC and one non-neoplastic lesion per case were microdissected from 15 patients with RCC. Polymerase chain reaction-amplified products were directly sequenced. Loss of heterozygosity (LOH) was examined at four sites of known polymorphism. Mutations of the Fas gene were detected in 3 RCC lesions from 3 (20%) of 15 cases. All mutations were point mutations, 2 missense and one silent, in exons 7 and 9. Non-neoplastic tissues never showed Fas gene mutations. Nine of 15 cases (60.0%) were heterozygous for one or more sites of the known biallelic polymorphisms, i.e., at nucleotides -1377, -670, 416, and 836. Two of these 9 cases showed LOH at promoter region -670. Mouse T-cell lymphoma cells transfected with missense mutated genes were resistant to apoptosis induced by anti-Fas antibody, indicating these to be loss-of-function mutations. The results of the present study suggest that Fas gene mutations play a role in the pathogenesis of RCC.
Insights
Fas gene mutations, including missense and silent point mutations, were identified in 20% of renal cell carcinoma (RCC) lesions, suggesting their role in RCC development and apoptosis resistance.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Renal cell carcinoma (RCC) expresses Fas antigen, making it potentially sensitive to Fas ligand-induced apoptosis.
- Fas gene mutations are hypothesized to contribute to RCC development.
Purpose of the Study:
- To investigate the presence and functional impact of Fas gene mutations in renal cell carcinoma.
- To determine if Fas gene mutations are associated with altered apoptosis sensitivity in RCC.
Main Methods:
- Genomic DNA was extracted from 15 RCC patient lesions and matched non-neoplastic tissues.
- Laser capture microdissection was used for precise tissue isolation.
- Polymerase chain reaction amplification and direct sequencing were employed to detect Fas gene mutations.
- Loss of heterozygosity (LOH) was assessed at four polymorphic sites.
- Functional analysis involved transfecting mouse T-cell lymphoma cells with mutated Fas genes.
Main Results:
- Fas gene mutations were found in 3 of 15 (20%) RCC lesions, specifically point mutations (2 missense, 1 silent) in exons 7 and 9.
- No Fas gene mutations were detected in non-neoplastic tissues.
- 60% of cases were heterozygous for polymorphisms in the Fas gene promoter region.
- Two cases exhibited LOH at the -670 promoter region.
- Transfected cells with missense Fas mutations showed resistance to anti-Fas antibody-induced apoptosis, indicating loss-of-function.
Conclusions:
- Fas gene mutations, particularly loss-of-function variants, are implicated in the pathogenesis of renal cell carcinoma.
- These mutations may contribute to the evasion of apoptosis in RCC cells.
- Further research into Fas signaling pathways could offer therapeutic targets for RCC.
