脱氧核糖核酸酶1在肾细胞癌中的作用及机制研究

doi:10.19586/j.2095-2341.2024.0048

生物技术进展 ›› 2024, Vol. 14 ›› Issue (3): 486-491.DOI: 10.19586/j.2095-2341.2024.0048

• 研究论文 • 上一篇

脱氧核糖核酸酶1在肾细胞癌中的作用及机制研究

曾艳(), 祝恒成, 杨康()

武汉大学人民医院泌尿外科，武汉 443002

收稿日期:2024-03-11 接受日期:2024-04-11 出版日期:2024-05-25 发布日期:2024-06-18
通讯作者: 杨康
作者简介:曾艳 E-mail: 2839541386@qq.com；
基金资助:
国家自然科学基金项目(82100703)

The Mechanism of DNASE1L3 in Renal Cell Carcinoma

Yan ZENG(), Hengcheng ZHU, Kang YANG()

Department of Urology，Renmin Hospital of Wuhan University，Wuhan 443002，China

Received:2024-03-11 Accepted:2024-04-11 Online:2024-05-25 Published:2024-06-18
Contact: Kang YANG

摘要/Abstract

摘要：

肾细胞癌（renal cell carcinoma，RCC）患者在手术后对放疗、化疗和靶向治疗均不敏感。前期研究证明，在肝癌中，脱氧核糖核酸酶1（deoxyribonuclease 1-like 3，DNASE1L3）可以分解包膜内的单链和双链DNA，通过弱化糖酵解的限速酶，抑制细胞周期、增殖和代谢。然而，DNASE1L3在肾癌中的作用和相关机制尚不清楚。从癌症基因组图谱数据库（the Cancer Genome Atlas，TCGA）中下载并分析了肾癌RNA测序数据；使用（Gene Expression Omnibus，GEO）数据库、人类蛋白质图谱数据库和免疫印迹验证DNASE1L3的表达水平；运用免疫相关数据库分析、划痕和侵袭实验，探究了DNASE1L3的作用和潜在机制。结果发现，在TCGA肾癌数据中，DNASE1L3表达量与患者的临床特征显著相关，且与患者生存期呈正相关；过表达DNASE1L3会抑制ACHN和786-O细胞的增殖和侵袭能力。结果表明，DNASE1L3可能成为肾癌患者诊断和治疗的潜在生物标志物。

关键词: 脱氧核糖核酸酶1, 肿瘤免疫, 肾癌, 免疫治疗

Abstract:

Renal cell carcinoma （RCC） patients are insensitive to radiotherapy， chemotherapy and targeted therapy after surgery. The previous study have proved that deoxyribonuclease 1-like 3 （DNASE1L3） could decompose both membrane-encapsulated single and double-stranded DNA， and suppressed the cell cycle， proliferation and metabolism by weakening rate-limiting enzymes of glycolysis in hepatocellular carcinoma. However， the mechanism underlying the relationship between DNASE1L3 and the tumor immune microenvironment in renal cell carcinoma is still unclear. The RNA sequencing data of RCC were downloaded and analyzed from the Cancer Genome Atlas （TCGA） database. The expression levels of DNASE1L3 were verified using the Gene Expression Omnibus （GEO） database， Human Protein Atlas database and Western bloting. The role and potential mechanism of DNASE1L3 were investigated with immune-related database analysis， wound healing， and invasion. The results revealed that DNASE1L3 expression was significantly correlated with the clinical characteristics of patients and positively correlated with patient survival. In addition， DNASE1L3 overexpression inhibited the proliferation and invasion capacities of ACHN and 786-O cells. This study proved that DNASE1L3 may be a promising potential biomarker for the diagnosis and treatment of RCC patients.

Key words: DNASE1L3, tumor immunity, renal cell carcinoma, immunotherapy

中图分类号:

Q291

曾艳, 祝恒成, 杨康. 脱氧核糖核酸酶1在肾细胞癌中的作用及机制研究[J]. 生物技术进展, 2024, 14(3): 486-491.

Yan ZENG, Hengcheng ZHU, Kang YANG. The Mechanism of DNASE1L3 in Renal Cell Carcinoma[J]. Current Biotechnology, 2024, 14(3): 486-491.

图/表 5

图1 DNASE1L3在肾癌中的表达水平A~C：DNASE1L3在GCE68417、GSE36895和GSE53757数据集中肾癌和癌旁正常组织的表达情况；D：免疫组化分析DNASE1L3在肾癌和癌旁正常组织的表达情况；E~F：免疫印迹分析DNASE1L3在肾癌细胞组和肾小管上皮细胞HK2的表达情况并进行定量分析。***表示与正常组相比在P<0.001水平上具有统计学意义。

Fig. 1 The expression level of DNASE1L3 in renal cell carcinoma

图2 DNASE1L3表达水平与肾癌患者临床特征关系A：Kaplan-Meier方法分析DNASE1L3在肾癌患者中的预后情况；B~D：Pearson相关性分析DNASE1L3与肾癌患者肿瘤体积、临床分期和病理分级的关系

Fig. 2 The relationship between DNASE1L3 expression levels and clinical characteristics of renal cancer patients

表1 TCGA KIRC队列中临床病理特征和DNASE1L3与总生存期的单变量和多变量分析

Table 1 Univariate and multivariate analysis of clinical and pathological features，DNASE1L3 and overall survival in the TCGA KIRC cohort

	单变量分析		多变量分析
	HR(95% CI)	P值	HR(95% CI)	P值
年龄	0.561(0.403~0.779)	<0.001	0.610(0.434~0.857)	0.004 3
性别	1.129(0.813~1.568)	0.469	1.107(0.786~1.558)	0.561 6
组织学分级	2.668(1.829~3.893)	<0.001	1.679(1.121~2.515)	0.012 0
病理分期	3.990(2.815~5.655)	<0.001	1.797(0.812~3.981)	0.148 4
病理学 T	3.240(2.329~4.507)	<0.001	1.250(0.632~2.474)	0.521 7
病理学 N	0.822(0.598~1.13)	0.228	0.867(0.619~1.215)	0.408 2
病理学 M	3.474(2.506~4.815)	<0.001	2.443(1.653~3.61)	<0.001 0
肿瘤尺寸	0.424(0.3~0.599)	<0.001	0.943(0.628~1.416)	0.778 3
肿瘤重量	0.562(0.407~0.776)	<0.001	0.557(0.394~0.788)	0.000 9
DNASE1L3	0.505(0.363~0.703)	<0.001	0.828(0.585~1.17)	0.028 5

图3 DNASE1L3对肾癌细胞增殖和侵袭能力的影响A：免疫印迹检测DNASE1L3在肾癌细胞株中的转染效率；B、E：伤口划痕实验分析转染对照组和转染组786-O以及ACHN在0和24 h的迁移距离及定量，放大倍数为40倍；C~D：transwell实验分析转染对照组和转染组786-O以及ACHN在0和24 h的迁徙能力及定量，放大倍数为100倍。***表示与转染对照组相比在P<0.001水平上具有统计学意义。

Fig. 3 Effects of DNASE1L3 on the proliferation and invasion ability of renal cancer cells

图4 Pearson相关性分析DNASE1L3与肾癌免疫微环境的关系注：*、**、***分别表示与DNASE1L3低表达组相比，在P<0.05、P<0.01和P<0.001水平上具有统计学意义。

Fig. 4 Pearson correlation analysis of the relationship between DNASE1L3 and the immune microenvironment of renal cancer

参考文献 12

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脱氧核糖核酸酶1在肾细胞癌中的作用及机制研究

The Mechanism of DNASE1L3 in Renal Cell Carcinoma

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