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Quantification of SSX mRNA expression in human bone and soft tissue tumors using nucleic acid sequence-based
Norifumi Naka1, Susumu Joyama, Yoshitane Tsukamoto
1Department of Orthopedic Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan.
Abstract:
The SSX family proteins have been considered new members of the cancer/testis antigens because of the restricted expression in testis among normal tissues and the activation in a wide range of cancers. Thus, they would be potential molecular targets for immunotherapeutic strategies. We have developed a competitive nucleic acid sequence-based amplification (NASBA) assay to analyze SSX mRNA expression in 211 bone and soft tissue tumors. The copy numbers of SSX mRNA per mug of total RNA in tumor tissues were widely distributed, ranging logarithmically from 0.6 to 6.6. We found that malignant tumors showed significantly higher expression of SSX mRNA than benign tumors (P < 0.0001). Further, SSX mRNA expression in stage III tumors was significantly higher than that in stage I or II tumors (P < 0.005). This NASBA assay was also more sensitive compared to immunohistochemistry using newly affinity-purified polyclonal antibody against SSX. Collectively, these results suggest that the SSX quantitative NASBA assay could provide useful information to select eligible patients for SSX-specific cancer vaccines.
Insights
Cancer/testis antigen SSX family proteins are promising targets for cancer vaccines. A new assay quantifies SSX mRNA in tumors, revealing higher expression in malignant and advanced-stage cancers, aiding patient selection for immunotherapy.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Cancer/testis antigens (CTAs) like SSX family proteins are expressed in tumors and testis.
- SSX proteins are potential targets for cancer immunotherapies due to their tumor-specific expression.
- Developing sensitive methods to detect SSX expression is crucial for targeted therapies.
Purpose of the Study:
- To develop and validate a quantitative assay for SSX mRNA expression in bone and soft tissue tumors.
- To investigate the correlation between SSX mRNA levels and tumor malignancy and stage.
- To compare the sensitivity of the developed assay with existing methods like immunohistochemistry.
Main Methods:
- Development of a competitive nucleic acid sequence-based amplification (NASBA) assay.
- Analysis of SSX mRNA expression in 211 bone and soft tissue tumor samples.
- Comparison of SSX mRNA levels between benign and malignant tumors, and across different tumor stages.
- Comparative analysis with immunohistochemistry (IHC) using a novel SSX antibody.
Main Results:
- SSX mRNA copy numbers varied logarithmically (0.6-6.6) across tumor samples.
- Malignant tumors exhibited significantly higher SSX mRNA expression than benign tumors (P < 0.0001).
- Tumors at stage III showed significantly higher SSX mRNA levels compared to stages I and II (P < 0.005).
- The NASBA assay demonstrated higher sensitivity than IHC.
Conclusions:
- The quantitative SSX NASBA assay is a sensitive tool for measuring SSX mRNA expression in tumors.
- Elevated SSX mRNA levels correlate with increased tumor malignancy and advanced stage.
- This assay can aid in identifying patients eligible for SSX-targeted cancer vaccines.
