College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2023 Oct 3;52(5):533-543. doi: 10.3724/zdxbyxb-2023-0256.
To investigate the effects and mechanisms of deubiquitinating enzyme Josephin domain containing 2 (JOSD2) on susceptibility of non-small cell lung carcinoma (NSCLC) cells to anti-cancer drugs.
The transcriptome expression and clinical data of NSCLC were downloaded from the Gene Expression Omnibus. Principal component analysis and limma analysis were used to investigate the deubiquitinating enzymes up-regulated in NSCLC tissues. Kaplan-Meier analysis was used to investigate the relationship between the expression of deubiquitinating enzymes and overall survival of NSCLC patients. Gene ontology enrichment and gene set enrichment analysis (GSEA) were used to analyze the activation of signaling pathways in NSCLC patients with high expression of . Gene set variation analysis and Pearson correlation were used to investigate the correlation between expression levels and DNA damage response (DDR) pathway. Western blotting was performed to examine the expression levels of JOSD2 and proteins associated with the DDR pathway. Immunofluorescence was used to detect the localization of JOSD2. Sulforhodamine B staining was used to examine the sensitivity of -knock-down NSCLC cells to DNA damaging drugs.
Compared with adjacent tissues, the expression level of was significantly up-regulated in NSCLC tissues (<0.05), and was significantly correlated with the prognosis in NSCLC patients (<0.05). Compared with the tissues with low expression of , the DDR-related pathways were significantly upregulated in NSCLC tissues with high expression of (all <0.05). In addition, the expression of was positively correlated with the activation of DDR-related pathways (all <0.01). Compared with the control group, overexpression of significantly promoted the DDR in NSCLC cells. In addition, DNA damaging agents significantly increase the nuclear localization of JOSD2, whereas depletion of significantly enhanced the sensitivity of NSCLC cells to DNA damaging agents (all <0.05).
Deubiquitinating enzyme JOSD2 may regulate the malignant progression of NSCLC by promoting DNA damage repair pathway, and depletion of significantly enhances the sensitivity of NSCLC cells to DNA damaging agents.
研究去泛素化酶 Josephin 结构域包含蛋白 2（JOSD2）对非小细胞肺癌（NSCLC）细胞对抗癌药物敏感性的影响及其作用机制。
从基因表达综合数据库中下载 NSCLC 的转录组表达和临床数据。采用主成分分析和 limma 分析方法研究 NSCLC 组织中上调的去泛素化酶。Kaplan-Meier 分析用于研究去泛素化酶的表达与 NSCLC 患者总生存期的关系。基因本体论富集和基因集富集分析（GSEA）用于分析高表达 NSCLC 患者信号通路的激活情况。基因集变异分析和 Pearson 相关性用于研究表达水平与 DNA 损伤反应（DDR）通路的相关性。Western blot 检测 JOSD2 及与 DDR 通路相关蛋白的表达水平。免疫荧光检测 JOSD2 的定位。磺酰罗丹明 B 染色检测 -敲低 NSCLC 细胞对 DNA 损伤药物的敏感性。
与相邻组织相比，NSCLC 组织中 （<0.05） 的表达水平显著上调，与 NSCLC 患者的预后显著相关（<0.05）。与低表达 相比，高表达 组织中 DDR 相关通路明显上调（均<0.05）。此外， 表达与 DDR 相关通路的激活呈正相关（均<0.01）。与对照组相比，过表达 显著促进 NSCLC 细胞的 DDR。此外，DNA 损伤剂显著增加 JOSD2 的核定位，而 耗竭则显著增强 NSCLC 细胞对 DNA 损伤剂的敏感性（均<0.05）。
去泛素化酶 JOSD2 可能通过促进 DNA 损伤修复通路调节 NSCLC 的恶性进展，而 耗竭则显著增强 NSCLC 细胞对 DNA 损伤剂的敏感性。
