Blocking the autocrine regulatory loop of Gankyrin/STAT3/CCL24/CCR3 impairs the progression and pazopanib resistance of clear cell renal cell carcinoma
Blocking the autocrine regulatory loop of Gankyrin/STAT3/CCL24/CCR3 impairs the progression and pazopanib resistance of clear cell renal cell carcinoma
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阻断 Gankyrin/STAT3/CCL24/CCR3 的自分泌调节环路会损害透明细胞肾细胞癌的进展和帕唑帕尼耐药性
DOI：
10.1038/s41419-020-2306-6
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发表时间：
2020-02-12
影响因子：
9
通讯作者：
Cui, Xingang
中科院分区：
文献类型：
--
作者：
Wang, Chao;Wang, Yuning;Cui, Xingang
透明细胞肾细胞癌（ccRCC）患者的预后不良是由于进展和靶向耐药，但潜在的分子机制需要进一步阐明。本研究在前人研究的基础上，探讨了Gankyrin在肾细胞癌中的生物学功能及其相关机制。为此，进行体外功能实验;皮下肿瘤形成、肺转移和原位ccRCC的体内模型;以及抗体芯片检测、co-IP、ChIP测定，以检查gankyrin在ccRCC中的生物学作用和分子机制。将200例五十六ccRCC患者随机分为训练组和验证组，以通过IHC和统计分析检查Gankyrin和其他标志物的预后价值。我们观察到，gankyrin过表达的ccRCC细胞系786-O和769-P表现出增殖、侵袭、迁移、致瘤性和帕唑帕尼抗性增加，凋亡减少，而gankyrin敲低则达到相反的结果。从机制上讲，gankyrin通过直接结合募集STAT 3，而STAT 3与CCL 24启动子的结合促进了其表达。反过来，自分泌CCL 24的增加增强了ccRCC中Gankyrin的表达和通过CCR 3激活STAT 3，形成正自分泌调节环。此外，体内实验结果显示，通过gankyrin敲低或用CCR 3抑制剂SB 328437处理来阻断正环逆转了对帕唑帕尼的抗性并抑制了ccRCC中的肺转移。此外，在ccRCC标本中观察到gankyrin和STAT 3或CCL 24表达之间的正相关性，并且通过将gankyrin和STAT 3或CCL 24表达与现有的临床预后指标（包括TNM分期和SSIGN评分）相结合来实现ccRCC患者预后的提高的准确性。总之，靶向gankyrin/STAT 3/CCL 24/CCR 3自分泌调节环可以作为晚期ccRCC患者的治疗方法，并且将gankyrin和STAT 3或CCL 24表达与当前临床指标相结合可以更好地预测ccRCC患者的预后。
The poor prognosis of clear-cell renal cell carcinoma (ccRCC) patients is due to progression and targeted drug resistance, but the underlying molecular mechanisms need further elucidation. This study examined the biological function and related mechanisms of gankyrin in ccRCC based on the results of our previous study. To this end, in vitro functional experiments; in vivo models of subcutaneous tumor formation, lung metastasis, and orthotopic ccRCC; and antibody chip detection, co-IP, ChIP assays were performed to examine the biological role and molecular mechanisms of gankyrin in ccRCC. Two hundred fifty-six ccRCC patients were randomly divided into training and validation cohorts to examine the prognostic value of gankyrin and other markers through IHC and statistical analyses. We observed that the gankyrin-overexpressing ccRCC cell lines 786-O and 769-P exhibited increased proliferation, invasion, migration, tumorigenicity, and pazopanib resistance and decreased apoptosis, while gankyrin knockdown achieved the opposite results. Mechanistically, gankyrin recruited STAT3 via direct binding, and STAT3 binding to the CCL24 promoter promoted its expression. Reciprocally, an increase in autocrine CCL24 enhanced the expression of gankyrin and STAT3 activation via CCR3 in ccRCC, forming a positive autocrine-regulatory loop. Furthermore, in vivo experimental results revealed that blocking the positive loop through gankyrin knockdown or treatment with the CCR3 inhibitor SB328437 reversed the resistance to pazopanib and inhibited lung metastasis in ccRCC. Moreover, a positive correlation between gankyrin and STAT3 or CCL24 expression in ccRCC specimens was observed, and improved accuracy for ccRCC patient prognosis was achieved by combining gankyrin and STAT3 or CCL24 expression with existing clinical prognostic indicators, including the TNM stage and SSIGN score. In summary, targeting the gankyrin/STAT3/CCL24/CCR3 autocrine-regulatory loop may serve as a remedy for patients with advanced ccRCC, and combining gankyrin and STAT3 or CCL24 expression with the current clinical indicators better predicts ccRCC patient prognosis.