Department of Integrative Oncology, Fudan University Shanghai Cancer Center, No. 270 Dongan Rd., Xuhui District, Shanghai, 200032, China.
Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, 200032, China.
Mol Med. 2023 Apr 4;29(1):47. doi: 10.1186/s10020-023-00648-z.
Protein kinases play a pivotal role in the malignant evolution of pancreatic cancer (PC) through mediating phosphorylation. Many kinase inhibitors have been developed and translated into clinical use, while the complex pathology of PC confounds their clinical efficacy and warrants the discovery of more effective therapeutic targets.
Here, we used the Gene Expression Omnibus (GEO) database and protein kinase datasets to map the PC-related protein kinase-encoding genes. Then, applying Gene Expression and Profiling Interactive Analysis (GEPIA), GEO and Human Protein Atlas, we evaluated gene correlation, gene expression at protein and mRNA levels, as well as survival significance. In addition, we performed protein kinase RIPK2 knockout and overexpression to observe effects of its expression on PC cell proliferation, migration and invasion in vitro, as well as cell apoptosis, reactive oxygen species (ROS) production and autophagy. We established PC subcutaneous xenograft and liver metastasis models to investigate the effects of RIPK2 knockout on PC growth and metastasis. Co-immunoprecipitation and immunofluorescence were utilized to explore the interaction between protein kinases RIPK2 and PRKCI. Polymerase chain reaction and immunoblotting were used to evaluate gene expression and protein phosphorylation level.
We found fourteen kinases aberrantly expressed in human PC and nine kinases with prognosis significance. Among them, RIPK2 with both serine/threonine and tyrosine activities were validated to promote PC cells proliferation, migration and invasion. RIPK2 knockout could inhibit subcutaneous tumor growth and liver metastasis of PC. In addition, RIPK2 knockout suppressed autophagosome formation, increased ROS production and PC cell apoptosis. Importantly, another oncogenic kinase PRKCI could interact with RIPK2 to enhance the phosphorylation of downstream NF-κB, JNK and ERK.
Paired protein kinases PRKCI-RIPK2 with multiple phosphorylation activities represent a new pathological mechanism in PC and could provide potential targets for PC therapy.
蛋白激酶通过介导磷酸化在胰腺癌（PC）的恶性演进中发挥关键作用。许多激酶抑制剂已经被开发并转化为临床应用，然而 PC 的复杂病理情况使它们的临床疗效复杂化，需要发现更有效的治疗靶点。
在这里，我们使用基因表达综合数据库（GEO）和蛋白激酶数据集来映射与 PC 相关的蛋白激酶编码基因。然后，我们应用基因表达和分析交互（GEPIA）、GEO 和人类蛋白质图谱，评估基因相关性、蛋白和 mRNA 水平的基因表达以及生存意义。此外，我们进行了蛋白激酶 RIPK2 敲除和过表达实验，观察其表达对 PC 细胞体外增殖、迁移和侵袭以及细胞凋亡、活性氧（ROS）产生和自噬的影响。我们建立了 PC 皮下移植瘤和肝转移模型，以研究 RIPK2 敲除对 PC 生长和转移的影响。我们利用免疫共沉淀和免疫荧光来探索蛋白激酶 RIPK2 和 PRKCI 之间的相互作用。聚合酶链反应和免疫印迹用于评估基因表达和蛋白磷酸化水平。
我们发现十四种激酶在人类 PC 中异常表达，其中九种激酶与预后相关。在这些激酶中，具有丝氨酸/苏氨酸和酪氨酸活性的 RIPK2 被证实可促进 PC 细胞的增殖、迁移和侵袭。RIPK2 敲除可抑制 PC 皮下肿瘤生长和肝转移。此外，RIPK2 敲除抑制自噬体形成，增加 ROS 产生和 PC 细胞凋亡。重要的是，另一种致癌激酶 PRKCI 可以与 RIPK2 相互作用，增强下游 NF-κB、JNK 和 ERK 的磷酸化。
具有多种磷酸化活性的配对蛋白激酶 PRKCI-RIPK2 代表了 PC 中的一种新的病理机制，并可为 PC 治疗提供潜在的靶点。
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