cDNA Expression Library Screening Revealed Novel Functional Genes Involved in Clear Cell Carcinogenesis of the Ovary in vitro
cDNA Expression Library Screening Revealed Novel Functional Genes Involved in Clear Cell Carcinogenesis of the Ovary in vitro
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cDNA表达文库筛选揭示了参与卵巢透明细胞体外癌变的新功能基因
DOI：
10.1080/01443615.2020.1716310
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发表时间：
2020
影响因子：
1.3
通讯作者：
Shiozawa T.
中科院分区：
文献类型：
--
作者：
Yamada Y;Miyamoto T;Higuchi S;Ono M;Kobara H;Asaka R;Ando H;Suzuki A;Shiozawa T.
为了确定参与卵巢透明细胞癌（CCC）发病机制的基因，使用cDNA表达文库进行功能筛选。我们从CCC细胞系（RMG-1）中提取mRNA，使用逆转录病毒载体建立cDNA文库，将该文库转染到小鼠NIH 3 T3细胞中，并对所得灶进行测序。对分离基因的组织类型特异性表达及其转化活性进行了评价。分离出7个基因。在这些基因中，SEC 61 B和DVL 1的mRNA表达在CCC中显著强于其他组织学类型（p<0.05）。免疫组织化学染色显示，SEC 61 B和C1 ORF 38的表达强于正常卵巢组织（p<0.05）。通过将SEC 61 B、C1 ORF 38和DVL 1转染到NIH 3 T3细胞中来确认病灶形成。本研究鉴定了新的基因，包括SEC 61 B，C1 ORF 38和DVL 1，参与CCC的发病机制。这些基因可能是CCC的额外治疗靶点。影响声明关于这个主题已经知道了什么？卵巢透明细胞癌（CCC）中有几种重要的遗传异常，包括ARID 1A和PIK 3CA突变。SEC 61 B、C1 ORF 38和DVL 1是新近发现的卵巢透明细胞癌发生的候选基因，这些发现对临床实践和/或进一步研究有何意义？使用cDNA表达文库进行功能筛选可能是鉴定致病功能基因的有用技术。利用该技术获得的信息可能为CCC提供新的治疗靶点。
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.