背景:胰腺导管腺癌易发生转移,导致生存时间短,生活质量低。LncRNAs是参与各种肿瘤进展的关键协调器。lncRNAFAM83H-AS1在PDAC进展中的潜在作用和机制仍然未知。
方法:要解决此问题,首先,我们对TCGA数据库中的lncRNA异常表达进行了分析,并确定FAM83H-AS1是促进胰腺癌细胞迁移的最有效方法.然后,FAM83H-AS1在患者血清中的表达水平,使用RT-qPCR检测肿瘤组织和PDAC细胞,通过FISH和RT-qPCR测定PDAC细胞中FAM83H-AS1的分布。接下来,进行了一系列体内和体外功能测定,以阐明FAM83H-AS1在PDAC细胞生长和转移中的作用。FAM83H-AS1和FAM83H(FAM83H-AS1的同源基因)之间的调控关系分别通过进行蛋白质和RNA降解测定来验证。进行Co-IP测定以探索FAM83H对β-连环蛋白的潜在调节机制。进行挽救测定以验证PDAC进展中FAM83H-AS1/FAM83H/β-连环蛋白轴的调节。
结果:FAM83H-AS1在PDAC患者的肿瘤组织和血清中高表达,并与较短的生存期相关。FAM83H-AS1显著促进增殖,PDAC细胞的侵袭和转移,通过保护FAM83HmRNA免于降解。重要的是,FAM83H蛋白在PDAC细胞中表现出与FAM83H-AS1相似的恶性功能,并能与β-连环蛋白结合。具体来说,FAM83H可以降低β-catenin的泛素化,并相应地激活Wnt/β-连环蛋白信号传导的效应基因。
结论:总的来说,FAM83H-AS1可以通过稳定FAM83H的mRNA表达来促进FAM83H的表达,允许FAM83H降低β-连环蛋白的泛素化,因此导致扩增的FAM83H-AS1/FAM83H/β-catenin信号轴促进PDAC进展。FAM83H-AS1可能是对抗PDAC的新的预后和治疗靶标。
BACKGROUND: Pancreatic ductal adenocarcinoma is prone to metastasis, resulting in short survival and low quality of life. LncRNAs are pivotal orchestrators that participate in various tumor progress. The underlying role and mechanism of lncRNA FAM83H-AS1 is still unknown in PDAC progression.
METHODS: To address this issue, firstly, we profiled and analyzed the aberrant lncRNA expression in TCGA database and identified FAM83H-AS1 as the most effective one in promoting the migration of pancreatic cancer cells. Then, the expression levels of FAM83H-AS1 in patient\'s serum, tumor tissues and PDAC cells were detected using RT-qPCR, and FAM83H-AS1 distribution in PDAC cells was determined by performing FISH and RT-qPCR. Next, a series of in vivo and in vitro functional assays were conducted to elucidate the role of FAM83H-AS1 in cell growth and metastasis in PDAC. The regulatory relationship between FAM83H-AS1 and FAM83H (the homologous gene of FAM83H-AS1) was verified by performing protein and RNA degradation assays respectively. Co-IP assays were performed to explore the potential regulatory mechanism of FAM83H to β-catenin. Rescue assays were performed to validate the regulation of the FAM83H-AS1/FAM83H/β-catenin axis in PDAC progression.
RESULTS: FAM83H-AS1 was highly expressed in the tumor tissues and serum of patients with PDAC, and was correlated with shorter survival. FAM83H-AS1 significantly promoted the proliferation, invasion and metastasis of PDAC cells, by protecting FAM83H mRNA from degradation. Importantly, FAM83H protein manifested the similar malignant functions as that of FAM83H-AS1 in PDAC cells, and could bind to β-catenin. Specifically, FAM83H could decrease the ubiquitylation of β-catenin, and accordingly activated the effector genes of Wnt/β-catenin signaling.
CONCLUSIONS: Collectively, FAM83H-AS1 could promote FAM83H expression by stabilizing its mRNA, allowing FAM83H to decrease the ubiquitylation of β-catenin, thus resulted in an amplified FAM83H-AS1/FAM83H/β-catenin signal axis to promote PDAC progression. FAM83H-AS1 might be a novel prognostic and therapeutic target for combating PDAC.