背景:鸟嘌呤核苷酸结合蛋白2(GNBP2)是一种GTP酶,在宿主免疫和某些类型的癌症中具有关键作用,但其在肾透明细胞癌(ccRCC)中的功能尚不完全清楚。
目的:这项工作探讨了GNBP2在ccRCC进展中的作用及其潜在的分子机制。
方法:使用两个公共人类癌症数据库TNMplot和TISIDB来分析GNBP2在ccRCC进展过程中的表达模式以及GNBP2表达与ccRCC患者临床特征之间的相关性。通过EdU染色确定ccRCC细胞中的GNBP2功能,流式细胞术,划痕伤口试验,transwell分析,和异种移植模型。使用qPCR评估基因表达,蛋白质印迹,免疫荧光染色,和免疫组织化学染色。
结果:GNBP2在ccRCC组织中的表达明显升高,并随着肿瘤分级的增加而逐渐升高。GNBP2表达较高的患者总生存时间较短。敲除GNBP2抑制肿瘤细胞增殖和细胞周期进程,降低迁移和侵袭能力,而GNBP2过表达表现出促肿瘤作用。通过RNA干扰沉默GNBP2可显着抑制荷瘤裸鼠的肿瘤生长,并降低增殖标志物Ki67。机械上,GNBP2下调抑制了STAT3信号转导,因为它降低了STAT3的磷酸化并调节了靶基因的表达,包括c-Myc,MMP2,N-钙粘蛋白,和E-cadherin.
结论:这些发现表明GNBP2通过调节STAT3信号转导促进ccRCC进展,表明GNBP2可能是ccRCC治疗的有希望的分子靶标。
Guanine nucleotide-binding protein 2 (GNBP2) is a GTPase that has critical roles in host immunity and some types of cancer, but its function in clear cell renal cell carcinoma (ccRCC) is not fully understood.
This work explored the role of GNBP2 in ccRCC progression and the underlying molecular mechanism.
Two public human cancer databases TNMplot and TISIDB were employed to analyze the expression pattern of GNBP2 during ccRCC progression and the correlation between GNBP2 expression and clinical features of ccRCC patients. GNBP2 functions in ccRCC cells were determined by EdU staining, flow cytometry, scratch wound assay, transwell assay, and xenograft model. Gene expression was evaluated using qPCR, Western blot, immunofluorescence staining, and immunohistochemical staining.
GNBP2 expression was significantly elevated in ccRCC tissues and increased gradually with the increasing tumor grades. Patients with higher GNBP2 expression had shorter overall survival times. Knockdown of GNBP2 suppressed tumor cell proliferation and cell cycle progression and reduced the capability of migration and invasion, while GNBP2 overexpression exhibited protumor effects. GNBP2 silencing by RNA interference significantly inhibited the tumor growth of tumor-bearing nude mice and decreased the proliferation marker Ki67. Mechanistically, GNBP2 downregulation suppressed the STAT3 signaling transduction, as it reduced the phosphorylation of STAT3 and modulated the expression of the target genes, including c-Myc, MMP2, N-cadherin, and E-cadherin.
These findings reveal that GNBP2 promotes ccRCC progression by regulating STAT3 signaling transduction, indicating that GNBP2 might be a promising molecular target for ccRCC therapy.