cDNA expression library screening revealed novel functional genes involved in clear cell carcinogenesis of the ovary in vitro
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Published on: January 12, 2020
Yasushi Yamada1, Tsutomu Miyamoto1, Shotaro Higuchi1
1Department of Obstetrics and Gynecology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Japan.
Abstract:
In order to identify genes involved in the pathogenesis of clear cell carcinoma of the ovary (CCC), functional screening using a cDNA expression library was performed. We extracted mRNA from a CCC cell line (RMG-1), established a cDNA library using a retroviral vector, transfected that library into mouse NIH3T3 cells and sequenced the resultant foci. The tissue-type specific expression of isolated genes and their transforming activities were evaluated. Seven genes were isolated. Of these genes, the mRNA expression of SEC61B and DVL1 is significantly stronger in CCC than in other histological types (p < .05). Immunohistochemical staining reveals the stronger expression of SEC61B and C1ORF38 than normal ovarian tissues (p < .05). Focus formation is confirmed by the transfection of SEC61B, C1ORF38, and DVL1 into NIH3T3 cells. The present study identified novel genes including SEC61B, C1ORF38, and DVL1, involved in the pathogenesis of CCC. These genes may be additional therapeutic targets for CCC.Impact statementWhat is already known on this subject? Several important genetic abnormalities, including ARID1A and PIK3CA mutations, have been reported in ovarian clear cell carcinoma (CCC).What the results of this study add? SEC61B, C1ORF38, and DVL1 were newly detected as candidate genes involved in ovarian clear cell carcinogenesis.What the implications are of these findings for clinical practice and/or further research? Functional screening using a cDNA expression library may be a useful technique to identify functional genes for pathogenesis. The information obtained using this technique may provide new therapeutic targets of CCC.
Insights
Researchers identified novel genes SEC61B, C1ORF38, and DVL1 involved in ovarian clear cell carcinoma (CCC) pathogenesis. These findings suggest new therapeutic targets for CCC treatment.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Ovarian clear cell carcinoma (CCC) has known genetic abnormalities like ARID1A and PIK3CA mutations.
- Identifying novel genes is crucial for understanding CCC pathogenesis and developing targeted therapies.
Purpose of the Study:
- To identify novel genes implicated in the pathogenesis of ovarian clear cell carcinoma (CCC) using functional screening.
- To evaluate the tissue-specific expression and transforming activity of isolated genes.
Main Methods:
- Functional screening using a retroviral cDNA expression library derived from an ovarian clear cell carcinoma cell line (RMG-1).
- Transfection of the library into mouse NIH3T3 cells, followed by sequencing of resultant foci.
- Evaluation of tissue-specific mRNA expression and immunohistochemical staining for candidate genes.
- Assessment of transforming activity via focus formation assays in NIH3T3 cells.
Main Results:
- Seven genes were isolated through functional screening.
- SEC61B and DVL1 showed significantly higher mRNA expression in CCC compared to other histological types (p < .05).
- SEC61B and C1ORF38 exhibited stronger expression in CCC compared to normal ovarian tissues (p < .05).
- SEC61B, C1ORF38, and DVL1 demonstrated focus-forming activity in NIH3T3 cells, indicating transforming potential.
Conclusions:
- SEC61B, C1ORF38, and DVL1 are newly identified candidate genes involved in ovarian clear cell carcinoma carcinogenesis.
- These genes represent potential novel therapeutic targets for CCC.
- Functional screening with cDNA expression libraries is an effective method for discovering genes relevant to cancer pathogenesis.
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