PDF(Pubmed)
Abstract：
Metastasis is the main death reason for triple-negative breast cancer (TNBC). Thus, identifying the driver genes associated with metastasis of TNBC is urgently needed. CRISPR screens have dramatically enhanced genome editing and made it possible to identify genes associated with metastasis. In this study, we identified and explored the crucial role of ras homolog family member V (RhoV) in TNBC metastasis. Here, we performed customized in vivo CRISPR screens targeting metastasis-related genes obtained from transcriptome analysis of TNBC. The regulatory role of RhoV in TNBC was validated using gain- or loss-of-function studies in vitro and in vivo. We further conducted immunoprecipitation and LC-MS/MS to explore the metastasis mechanism of RhoV. In vivo functional screens identified RhoV as a candidate regulator involved in tumor metastasis. RhoV was frequently upregulated in TNBC and correlated with poor survival. Knockdown of RhoV significantly suppressed cell invasion, migration, and metastasis both in vitro and in vivo. In addition, we provided evidence that p-EGFR interacted with RhoV to activate the downstream signal pathway of RhoV, thereby promoting tumor metastasis. We further confirmed that this association was dependent on GRB2 through a specific proline-rich motif in the N-terminus of RhoV. This mechanism of RhoV is unique, as other Rho family proteins lack the proline-rich motif in the N-terminus.
摘要：
转移是三阴性乳腺癌（TNBC）的主要死亡原因。因此，迫切需要鉴定与TNBC转移相关的驱动基因。CRISPR筛选显着增强了基因组编辑，并使鉴定与转移相关的基因成为可能。在这项研究中,我们确定并探索了Ras同源家族成员V(RhoV)在TNBC转移中的关键作用。这里,我们针对从TNBC转录组分析获得的转移相关基因进行了定制的体内CRISPR筛选.使用体外和体内的功能增益或功能丧失研究验证了RhoV在TNBC中的调节作用。我们进一步进行了免疫沉淀和LC-MS/MS来探讨RhoV的转移机制。体内功能筛选将RhoV鉴定为涉及肿瘤转移的候选调节因子。RhoV在TNBC中经常上调，并与低生存率相关。敲除RhoV显著抑制细胞侵袭,体内和体外的迁移和转移。此外,我们提供了p-EGFR与RhoV相互作用激活RhoV下游信号通路的证据，从而促进肿瘤转移。我们进一步证实，这种关联通过RhoVN末端的特定富含脯氨酸的基序依赖于GRB2。RhoV的这种机制是独特的，因为其他Rho家族蛋白在N端缺乏富含脯氨酸的基序。
