Identification of P-Rex1 in the Regulation of Liver Cancer Cell Proliferation and Migration via HGF/c-Met/Akt Pathway.
Identification of P-Rex1 in the Regulation of Liver Cancer Cell Proliferation and Migration via HGF/c-Met/Akt Pathway.
复制标题
鉴定 P-Rex1 通过 HGF/c-Met/Akt 途径调节肝癌细胞增殖和迁移。
DOI：
10.2147/ott.s265592
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发表时间：
2020
影响因子：
4
通讯作者：
Sun G
中科院分区：
文献类型：
--
作者：
Qiu W;Chang Y;Liu J;Yang X;Yu Y;Li J;Liang Q;Sun G
rho - gtpase及其激活剂鸟嘌呤核苷酸交换因子（gef）越来越被认为是致癌信号传导的重要介质。P-Rex1是Rac小GTPase gef的Dbl家族成员之一，虽然已知它有助于癌细胞的迁移，但其在肝癌中的确切作用及其潜在机制尚不清楚。分析基因表达综合数据库（Gene Expression Omnibus database， GEO）的公共数据集和临床肝癌样本，探讨P-Rex1的表达。利用重组慢病毒转染系统建立P-Rex1敲低和过表达细胞系。采用BrdU和菌落形成试验测定细胞活力。迁移能力分析使用跨井迁移试验和体外伤口愈合试验。建立皮下移植瘤裸鼠，观察P-Rex1对体内肿瘤发生的影响。通过Western blot和免疫共沉淀法检测P-Rex1在肝癌发生中的作用。从临床资料比较，我们在肝癌肿瘤中发现了内源性P-Rex1的诱导表达。作为对HGF处理的反应，p - rex1敲低细胞在体外表现出增殖和迁移减少，以及体内异种移植物肿瘤生长减少。P-Rex1过表达促进肝癌细胞增殖和迁移。P-Rex1主要作为GPCR信号传导的下游效应因子。本研究表明，P-Rex1的下调通过降低酪氨酸激酶受体c-Met的磷酸化，导致Akt和Erk1/2的磷酸化显著降低。此外，在HGF处理后，P-Rex1和c-Met之间存在物理关联，这表明P-Rex1可能参与了HGF/c-Met信号通路。这些结果支持P-Rex1在肝癌中的新作用，这表明靶向P-Rex1可能为肝癌治疗提供潜在的策略。
Rho-GTPases and their activators, guanine nucleotide exchange factors (GEFs), are increasingly being recognized as essential mediators of oncogenic signaling. Although it is known that P-Rex1, a member of the Dbl family of GEFs for the Rac small GTPase, contributes to the migration of cancer cells, its exact role in liver cancer and the underlying mechanisms remain unclear. Public datasets from the Gene Expression Omnibus database (GEO) and clinical liver cancer samples were analyzed to explore the expression of P-Rex1. P-Rex1 knockdown and overexpression cell lines were established using a recombinant lentiviral transfection system. BrdU and colony formation assays were performed to determine cell viability. Migratory capacity was analyzed using a transwell migration assay and an in vitro wound-healing assay. Nude mice bearing subcutaneous xenograft tumors were established to determine the effects of P-Rex1 on tumorigenesis in vivo. The role of P-Rex1 in hepatocarcinogenesis was determined through Western blot and co-immunoprecipitation. Induced expression of endogenous P-Rex1 was identified in liver cancer tumors when compared with adjacent nonmalignant tissues from clinical data. In response to HGF treatment, P-Rex1-knockdown cells displayed reduced proliferation and migration in vitro as well as reduced xenograft tumor growth in vivo. Overexpression of P-Rex1 promoted liver cancer cell proliferation and migration. P-Rex1 primarily acts as a downstream effector of GPCR signaling. This study demonstrated that downregulation of P-Rex1 led to a significant decrease in the phosphorylation of Akt and Erk1/2 by reducing the phosphorylation of the tyrosine kinase receptor c-Met. Furthermore, a physical association between P-Rex1 and c-Met was observed after HGF treatment, suggesting that P-Rex1 may be involved in the HGF/c-Met signaling pathway. These results support the role of P-Rex1 as a novel player in liver cancer, which suggest that targeting P-Rex1 may provide a potential strategy for liver cancer treatment.