2022年3月，复旦大学(Fudan University)Houbao Liu老师研究团队在《Research Square》上发表论文：

“GPRIN1 promotes gallbladder cancer progression through the CDK1 pathway”

“GPRIN1通过CDK1途径促进胆囊癌的进展”

Abstract：

Abstract

Purpose Recent studies have shown that G protein-regulated inducer of neurite outgrowth 1 (GPRIN1) regulates cancer proliferation and migration. However, there is limited research regarding its role in gallbladder cancer (GBC). We aimed to investigate the mechanism by which GPRIN1 promotes GBC cell proliferation and migration.

Methods Immunohistochemistry was performed to visualize in situ GPRIN1 expression in 65 human GBC tissues and 59 corresponding normal tissues. The Kaplan–Meier method was used to analyze the correlation between GPRIN1 and overall survival. Cell proliferation (Celigo and MTT), apoptosis (FACS), and cell migration and invasion assays and a subcutaneous xenograft tumor model were used to define the effects of GPRIN1 downregulation in GBC cells in vitro and in vivo. Coimmunoprecipitation assays, mass spectrometry and Western blotting were performed to identify GPRIN1-interacting proteins. Cell proliferation, MTT and Transwell assays were performed to verify the functional recovery effect of cyclin-dependent protein kinase 1 (CDK1) on GPRIN1. Proteomic analysis of differentially expressed proteins and/or phosphorylated peptides was performed.

Results GPRIN1 is upregulated in GBC tumor tissues and high GPRIN1 expression is positively correlated with a poor prognosis in GBC patients. High GPRIN1 expression reinforced the proliferative and invasive capacities of GBC in vitro and in vivo. Coimmunoprecipitation assays, mass spectrometry and Western blotting validated the direct interaction between GPRIN1 and CDK1. Cell proliferation, MTT and Transwell assays suggested that CDK1 exerts a functional recovery effect on GPRIN1. Proteomic analysis revealed the involvement of CDK1 in many biological processes and cellular pathways.

Conclusions GPRIN1 promotes the invasion and metastasis of GBC cells via CDK1 pathway activation.

摘要：

摘要

近期研究表明，G蛋白调控的神经突生长诱导因子1 (GPRIN1)调控肿瘤的增殖和迁移。然而，关于其在胆囊癌(GBC)中的作用的研究有限。我们的目的是研究GPRIN1促进GBC细胞增殖和迁移的机制。

方法采用免疫组化方法观察65例人GBC组织和59例正常组织中GPRIN1的原位表达。采用Kaplan-Meier法分析GPRIN1与总生存率的相关性。通过细胞增殖(Celigo和MTT)、细胞凋亡(FACS)、细胞迁移和侵袭试验以及皮下异种移植肿瘤模型来确定GPRIN1下调对GBC细胞的体外和体内影响。采用共免疫沉淀法、质谱法和Western blotting鉴定gprin1相互作用蛋白。细胞增殖、MTT和Transwell实验验证细胞周期蛋白依赖性蛋白激酶1 (cyclin-dependent protein kinase 1, CDK1)对GPRIN1的功能恢复作用。对差异表达蛋白和/或磷酸化肽进行蛋白质组学分析。

结果GPRIN1在GBC肿瘤组织中表达上调，GPRIN1高表达与GBC患者预后不良呈正相关。GPRIN1的高表达增强了GBC在体外和体内的增殖和侵袭能力。共免疫沉淀法、质谱法和Western blotting验证了GPRIN1和CDK1之间的直接相互作用。细胞增殖、MTT和Transwell实验表明，CDK1对GPRIN1具有功能恢复作用。蛋白质组学分析显示CDK1参与许多生物学过程和细胞通路。

结论GPRIN1通过激活CDK1通路促进GBC细胞的侵袭和转移。

该论文中，GBC-SD、NOZ人GBC细胞系和293T人胚胎肾细胞是使用Ausbian进口特级胎牛血清完成的。