Chen Jing, Lou Weiyang, Ding Bisha, Wang Xian
Department of Medical Oncology, Sir Run Run Shaw Hospital, Medical School of Zhejiang University, Zhejiang Province, Hangzhou 313100, China.
First Affiliated Hospital of Jiaxing University, Zhejiang Province, Jiaxing 314000, China.
Aging (Albany NY). 2019 Aug 13;11(15):5666-5688. doi: 10.18632/aging.102152.
Growing studies have reported that pseudogenes play key roles in multiple human cancers. However, expression and roles of pseudogenes in renal cell carcinoma remains absent.
31 upregulated and 16 downregulated pseudogenes were screened. Higher expression of DUXAP8 and DUXAP9 indicated poorer prognosis of kidney cancer. 33 and 5 miRNAs were predicted to potentially binding to DUXAP8 and DUXAP9, respectively. miR-29c-3p was identified as the most potential binding miRNAs of DUXAP8 and DUXAP9 based on expression, survival and correlation analyses. 254 target genes of miR-29c-3p were forecast. 47 hub genes with node degree >= 10 were identified. Subsequent analysis for the top 10 hub genes demonstrated that COL1A1 and COL1A2 may be two functional targets of DUXAP8 and DUXAP9. Expression of DUXAP8, DUXAP9, COL1A1 and COL1A2 were significantly increased in cancer samples compared to normal controls while miR-29c-3p expression was decreased. Luciferase reporter assay revealed that miR-29c-3p could directly bind to DUXAP8, DUXAP9, COL1A1 and COL1A2. Functional experiments showed that DUXAP8 and DUXAP9 enhanced but miR-29c-3p weakened growth of renal cell carcinoma.
In conclusion, upregulated DUXAP8 and DUXAP9 promote growth of renal cell carcinoma and serve as two promising prognostic biomarkers.
Dysregulated pseudogenes were obtained by dreamBase and GEPIA. The binding miRNAs of pseudogene and targets of miRNA were predicted using starBase and miRNet. Kaplan-Meier plotter was utilized to perform survival analysis, and Enrichr database was introduced to conduct functional enrichment analysis. Hub genes were identified through STRING and Cytoscape. qRT-PCR, luciferase reporter assay, cell counting assay and colony formation assay were performed to validate analytic results.
越来越多的研究报道假基因在多种人类癌症中发挥关键作用。然而，假基因在肾细胞癌中的表达及作用仍不清楚。
筛选出31个上调和16个下调的假基因。DUXAP8和DUXAP9的高表达表明肾癌预后较差。分别预测到33个和5个可能与DUXAP8和DUXAP9结合的miRNA。基于表达、生存和相关性分析，miR-29c-3p被确定为DUXAP8和DUXAP9最具潜力的结合miRNA。预测出miR-29c-3p的254个靶基因。鉴定出47个节点度≥10的枢纽基因。对前10个枢纽基因的后续分析表明，COL1A1和COL1A2可能是DUXAP8和DUXAP9的两个功能靶点。与正常对照相比，癌组织样本中DUXAP8、DUXAP9、COL1A1和COL1A2的表达显著增加，而miR-29c-3p的表达降低。荧光素酶报告基因实验表明，miR-29c-3p可直接与DUXAP8、DUXAP9、COL1A1和COL1A2结合。功能实验表明，DUXAP8和DUXAP9促进肾细胞癌生长，而miR-29c-3p则抑制其生长。
总之，上调的DUXAP8和DUXAP9促进肾细胞癌生长，是两个有前景的预后生物标志物。
通过dreamBase和GEPIA获取失调的假基因。使用starBase和miRNet预测假基因的结合miRNA和miRNA的靶标。利用Kaplan-Meier绘图工具进行生存分析，并引入Enrichr数据库进行功能富集分析。通过STRING和Cytoscape鉴定枢纽基因。进行qRT-PCR、荧光素酶报告基因实验、细胞计数实验和集落形成实验以验证分析结果。
