Abstract：
The cyclin-dependent kinase inhibitor 3 (CDKN3) gene, is over expressed in renal cell carcinoma (RCC). However, the cell biology functions of RCC are not well understood. The present study aimed to verify the ability of CDKN3 to promote the proliferation and migration of RCC through in vitro experiments. Subsequently, the clinical prognostic effects were analyzed using The Cancer Genome Atlas (TCGA; https://www.cancer.gov/) and Gene Expression Omnibus (GEO; https://www.ncbi.nlm.nih.gov/geo/). The chelators, di-2-pyridylketone 4,4-dimethyl-3-thiosemicarbazone (Dp44mT), an analogue of the anti-tumor agent, were screened through bioinformatics analysis. The expression of CDKN3 is positively correlated with the IC50 of Dp44mT. In two RCC cell lines, 786-0 and Caki-1, we conducted small interfering RNA (siRNA) knockdown of CDKN3 and overexpression of CDKN3 by transfection plasmid. Subsequently, we administered Dp44mT to examine the resulting alterations in cell proliferation, migration, and apoptosis, thereby elucidating the role of CDKN3 and Dp44mT in these processes. The results of the experiment revealed a positive association between CDKN3 expression and the proliferation of RCC cell lines. Down-regulating CDKN3 significantly increased the apoptosis rate and inhibited cell migration in 786-0 and Caki-1 cells. Furthermore, bioinformatics analysis revealed a high expression of CDKN3 in RCC and a negative association between CDKN3 expression and survival. Gene set enrichment analysis (GSEA) revealed a significant association between high CDKN3 expression and the cell cycle pathway. Furthermore, we identified Dp44mT as a drug highly correlated with CDKN3 through the database. Subsequent addition of Dp44mT resulted in similar findings to those observed upon CDKN3 knockdown. Our findings have important implications for the diagnosis and treatment of CDKN3 in RCC. Additionally, Dp44mT is likely to be a promising candidate for future clinical applications.
摘要：
细胞周期蛋白依赖性激酶抑制剂3（CDKN3）基因，在肾细胞癌(RCC)中过度表达。然而,RCC的细胞生物学功能尚不清楚。本研究旨在通过体外实验验证CDKN3促进RCC增殖和迁移的能力。随后，使用癌症基因组图谱(TCGA;https://www.cancer.gov/)和基因表达综合(GEO;https://www.ncbi.nlm.nih.gov/geo/)。螯合剂,二-2-吡啶基酮4,4-二甲基-3-氨基硫脲(Dp44mT),抗肿瘤剂的类似物，通过生物信息学分析进行筛选。CDKN3的表达与Dp44mT的IC50呈正相关。在两个RCC细胞系中，786-0和Caki-1，我们通过转染质粒进行了CDKN3的小干扰RNA（siRNA）敲低和CDKN3的过表达。随后，我们使用Dp44mT来检查细胞增殖的结果变化，迁移,和细胞凋亡，从而阐明CDKN3和Dp44mT在这些过程中的作用。实验结果表明，CDKN3表达与RCC细胞系的增殖之间存在正相关。下调CDKN3可显著提高786-0和Caki-1细胞的凋亡率并抑制细胞迁移。此外，生物信息学分析显示CDKN3在RCC中高表达，CDKN3表达与生存之间呈负相关。基因集富集分析（GSEA）揭示了高CDKN3表达与细胞周期途径之间的显着关联。此外，我们通过数据库确定Dp44mT是与CDKN3高度相关的药物.随后添加Dp44mT导致与在CDKN3敲低时观察到的相似的发现。我们的发现对RCC中CDKN3的诊断和治疗具有重要意义。此外,Dp44mT可能是未来临床应用的有希望的候选者。
