METTL3 and IGF2BP1-Mediated m6A Modification of ZHX2 Promotes Tumor Property of Renal Cell Carcinoma.

IF 2.3 4区医学 Q2 PERIPHERAL VASCULAR DISEASE

Kidney & blood pressure research Pub Date : 2024-01-01 Epub Date: 2024-08-19 DOI:10.1159/000540483

Bangming Xiao, Yalan Li, Yong Yang, Congbo Chen, Shide Gong, Hao Li, Qisheng Yao, Li Wang

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引用次数: 0

Abstract

Introduction: Renal cell carcinoma (RCC) is a common type of kidney cancer with limited treatment options and a high mortality rate. Therefore, it is essential to understand the role and mechanism of key genes in RCC development and progression. This study aimed to analyze the role of zinc fingers and homeoboxes 2 (ZHX2) in RCC and the underlying mechanism.

Methods: RNA expression was analyzed by quantitative real-time polymerase chain reaction, while protein expression was analyzed by Western blotting assay and immunohistochemistry assay. Cell viability was evaluated using CCK-8 assay, and cell proliferation was assessed by EdU assay. The rate of cell apoptosis was quantified by flow cytometry. Transwell assays were conducted to analyze cell migration and invasion. The sphere formation assay was performed to assess the formation of microspheres. Additionally, m6A RNA immunoprecipitation assay and RNA immunoprecipitation assay were utilized to investigate the relationship between ZHX2 and two proteins, methyltransferase like 3 (METTL3) and insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1). The stability of ZHX2 mRNA was analyzed through the Actinomycin D assay. Furthermore, a xenograft mouse model assay was conducted to analyze the effect of ZHX2 overexpression and METTL3 silencing on RCC cell tumor properties in vivo.

Results: ZHX2 expression was upregulated in both RCC tissues and cells when compared with healthy renal tissues and human renal cortex proximal convoluted tubule epithelial cells. Depletion of ZHX2 inhibited RCC cell proliferation, migration, invasion, and spheroid-forming capacity but promoted cell apoptosis. Moreover, it was found that METTL3-mediated m6A methylation of ZHX2 and IGF2BP1 also stabilized ZHX2 through m6A methylation modification. Furthermore, ZHX2 overexpression showed a potential for attenuating the effects induced by METTL3 silencing and counteracted the inhibitory effect of METTL3 depletion on tumor formation in vivo.

Conclusion: METTL3 and IGF2BP1-mediated m6A modification of ZHX2 promoted RCC progression. The finding suggests that ZHX2 may serve as a potential therapeutic target in RCC, providing valuable insights for future clinical interventions.

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METTL3 和 IGF2BP1 介导的 ZHX2 m6A 修饰可促进肾细胞癌的肿瘤特性。

导言：肾细胞癌（RCC）是一种常见的肾癌，治疗方法有限，死亡率高。因此，了解关键基因在 RCC 发生和发展中的作用和机制至关重要。本研究旨在分析锌指和同源染色体 2（ZHX2）在 RCC 中的作用及其内在机制：方法：采用实时定量聚合酶链式反应分析 RNA 表达，采用 Western 印迹分析和免疫组化分析蛋白质表达。用 CCK-8 法评估细胞活力，用 EdU 法评估细胞增殖。细胞凋亡率通过流式细胞术进行量化。透孔试验用于分析细胞迁移和侵袭。球形成试验用于评估微球的形成。此外，还利用 m6A RNA 免疫沉淀试验和 RNA 免疫沉淀试验研究了 ZHX2 与两种蛋白质（类似甲基转移酶 3 (METTL3)和胰岛素样生长因子 2 mRNA 结合蛋白 1 (IGF2BP1)）之间的关系。通过放线菌素 D 试验分析了 ZHX2 mRNA 的稳定性。此外，还进行了异种移植小鼠模型试验，以分析 ZHX2 过表达和 METTL3 沉默对 RCC 细胞体内肿瘤特性的影响：结果：与健康肾组织和人肾皮质近曲小管上皮细胞相比，ZHX2在RCC组织和细胞中均表达上调。抑制 ZHX2 可抑制 RCC 细胞的增殖、迁移、侵袭和球形细胞形成能力，但可促进细胞凋亡。研究还发现，METTL3介导了ZHX2的m6A甲基化，IGF2BP1也通过m6A甲基化修饰稳定了ZHX2。此外，ZHX2的过表达显示出减弱METTL3沉默诱导效应的潜力，并抵消了METTL3耗竭对体内肿瘤形成的抑制作用：结论：METTL3和IGF2BP1介导的ZHX2 m6A修饰促进了RCC的进展。这一发现表明，ZHX2可能是RCC的潜在治疗靶点，为未来的临床干预提供了宝贵的启示。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Kidney & blood pressure research 医学-泌尿学与肾脏学

CiteScore

4.80

自引率

3.60%

发文量

审稿时长

6-12 weeks

期刊介绍： This journal comprises both clinical and basic studies at the interface of nephrology, hypertension and cardiovascular research. The topics to be covered include the structural organization and biochemistry of the normal and diseased kidney, the molecular biology of transporters, the physiology and pathophysiology of glomerular filtration and tubular transport, endothelial and vascular smooth muscle cell function and blood pressure control, as well as water, electrolyte and mineral metabolism. Also discussed are the (patho)physiology and (patho) biochemistry of renal hormones, the molecular biology, genetics and clinical course of renal disease and hypertension, the renal elimination, action and clinical use of drugs, as well as dialysis and transplantation. Featuring peer-reviewed original papers, editorials translating basic science into patient-oriented research and disease, in depth reviews, and regular special topic sections, ''Kidney & Blood Pressure Research'' is an important source of information for researchers in nephrology and cardiovascular medicine.