CCNB1 是一种新型的预后生物标志物，可促进肾母细胞瘤的增殖、迁移和侵袭。
CCNB1 is a novel prognostic biomarker and promotes proliferation, migration and invasion in Wilms tumor.
机构信息
Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of Pediatrics, National Clinical Research Center for Child Health and Disorders, International Science and Technology Cooperation Base of Child Development and Critical Disorders, Children's Hospital of Chongqing Medical University, Chongqing, P.R. China.
Department of Urology, Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Children's Hospital of Chongqing Medical University, Chongqing, P.R. China.
出版信息
BMC Med Genomics. 2023 Aug 17;16(1):189. doi: 10.1186/s12920-023-01627-3.
BACKGROUND
Wilms tumour (WT) is a mixed type of embryonal tumour that usually occurs in early childhood. However, our knowledge of the pathogenesis or progression mechanism of WT is inadequate, and there is a scarcity of beneficial therapeutic strategies.
METHODS
High-throughput RNA sequencing was employed in this study to identify differentially expressed genes (DEGs) in clinical tumor samples and matching normal tissues. The STRING database was utilized to build a protein-protein interaction (PPI) network, and the Cytohubba method was used to identify the top 10 highly related HUB genes. Then, the key genes were further screened by univariate COX survival analysis. Subsequently, the XCELL algorithm was used to evaluate the tumour immune infiltration. RT-PCR, WB, and IF were used to verify the expression level of key genes in clinical tissues and tumour cell lines. Finally, the function of the key gene was further verified by loss-of-function experiments.
RESULTS
We initially screened 1612 DEGs, of which 1030 were up-regulated and 582 were down-regulated. The GO and KEGG enrichment analysis suggested these genes were associated with 'cell cycle', 'DNA replication'. Subsequently, we identified 10 key HUB genes, among them CCNB1 was strongly related to WT patients' overall survival. Multiple survival analyses showed that CCNB1 was an independent indicator of WT prognosis. Thus, we constructed a nomogram of CCNB1 combined with other clinical indicators. Single gene GSEA and immune infiltration analysis revealed that CCNB1 was associated with the degree of infiltration or activation status of multiple immune cells. TIDE analysis indicated that this gene was correlated with multiple key immune checkpoint molecules and TIDE scores. Finally, we validated the differential expression level of CCNB1 in an external gene set, the pan-cancer, clinical samples, and cell lines. CCNB1 silencing significantly inhibited the proliferation, migration, and invasive capabilities of WIT-49 cells, also, promoted apoptosis, and in turn induced G2 phase cell cycle arrest in loss-of-function assays.
CONCLUSION
Our study suggests that CCNB1 is closely related to WT progression and prognosis, and serves as a potential target.
背景
Wilms 瘤（WT）是一种混合胚胎性肿瘤，通常发生在儿童早期。然而，我们对 WT 的发病机制或进展机制的认识还不够充分，并且缺乏有益的治疗策略。
方法
本研究采用高通量 RNA 测序技术，鉴定临床肿瘤样本和匹配的正常组织中的差异表达基因（DEGs）。利用 STRING 数据库构建蛋白质-蛋白质相互作用（PPI）网络，采用 Cytohubba 方法筛选前 10 个高度相关的 HUB 基因。然后，通过单变量 COX 生存分析进一步筛选关键基因。随后，采用 XCELL 算法评估肿瘤免疫浸润。采用 RT-PCR、WB 和 IF 验证关键基因在临床组织和肿瘤细胞系中的表达水平。最后，通过功能丧失实验进一步验证关键基因的功能。
结果
我们初步筛选出 1612 个 DEGs，其中 1030 个上调，582 个下调。GO 和 KEGG 富集分析表明这些基因与“细胞周期”、“DNA 复制”有关。随后，我们鉴定出 10 个关键的 HUB 基因，其中 CCNB1 与 WT 患者的总生存密切相关。多项生存分析表明，CCNB1 是 WT 预后的独立指标。因此，我们构建了一个 CCNB1 与其他临床指标相结合的列线图。单基因 GSEA 和免疫浸润分析表明，CCNB1 与多种免疫细胞的浸润程度或激活状态有关。TIDE 分析表明，该基因与多个关键免疫检查点分子和 TIDE 评分相关。最后，我们验证了 CCNB1 在外部基因集、泛癌、临床样本和细胞系中的差异表达水平。CCNB1 沉默显著抑制 WIT-49 细胞的增殖、迁移和侵袭能力，同时促进细胞凋亡，进而在功能丧失实验中诱导 G2 期细胞周期停滞。
结论
本研究表明，CCNB1 与 WT 的进展和预后密切相关，可作为潜在的治疗靶点。
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