背景:转移是结直肠癌(CRC)患者死亡的主要原因。癌症已扩散到远处的CRC患者的5年生存率为13.5%。CRC转移的最常见部位是肝和肺。CRC转移性疾病的主要治疗方法是手术,但它的好处是有限的。
目的:本研究旨在揭示小檗碱对CRC细胞继发归巢形成转移灶的调控机制。这比以往直接研究CRC细胞的迁移和转移特征更有价值。
方法:在本研究中,我们使用小檗碱治疗后差异表达基因的功能富集分析,并通过WGCNA研究与CRC转移相关的共表达模块。还进行了重要模块的PPI和生存分析。通过一系列体外和体内实验研究了小檗碱在CRC肺和肝转移中的生物学功能:MTT,集落形成和小鼠尾静脉注射。我们扫描了HEY2蛋白的整个胞外域,用于与小檗碱的自动对接分析。
结果:我们发现小檗碱治疗后差异表达基因(DEGs)与癌症进展和转移相关通路有关。通过WGCNA分析,检测到4个癌症进展和转移相关模块.经过PPI和生存分析,我们鉴定并验证了HEY2为hub基因,转移期的高表达和低生存率。功能上,小檗碱抑制存活,CRC细胞在体外和体内的侵袭和迁移。机械上,小檗碱治疗下调HEY2、转移相关蛋白E-cadherin的表达,间充质上皮转化(MET)过程中的β-catenin和CyclinD1。小檗碱与HEY2表现出显著的交互作用,小檗碱在残基HIS-99界面与HEY2蛋白结合,氢键距离为1.9A。
结论:我们发现小檗碱可以显著抑制CRC肺和肝转移MET过程中hub基因HEY2和转移相关蛋白E-cadherin、β-catenin和CyclinD1的表达。总的来说,HEY2是CRC转移中预后和分子特征的有希望的候选生物标志物。
BACKGROUND: Metastasis is the leading cause of death in patients with colorectal cancer (CRC). The 5-year survival rate of CRC patients in whom the cancer has spread to distant sites is 13.5%. The most common sites of CRC metastasis are liver and lung. The principal therapies for CRC metastatic disease are surgery, but its benefits are limited.
OBJECTIVE: This study aimed to reveal the regulatory mechanism of berberine on secondary homing of CRC cells to form metastatic focus. This was more valuable than the previous direct study of the migration and metastasis characteristics of CRC cells.
METHODS: In this study, we used the functional enrichment analysis of differentially expressed genes after berberine treatment and investigated co-expression modules related with CRC metastasis by WGCNA. PPI and survival analyses of significant modules were also conducted. The biological functions of berberine in CRC lung and liver metastasis were investigated by a series of in vitro and in vivo experiments: MTT, colony formation and mouse tail vein injection. And we scanned through the entire extracellular domain of
HEY2 protein for autodocking analysis with berberine.
RESULTS: We found the differentially expressed genes (DEGs) after berberine treatment were related with cancer progression and metastasis related pathways. Through WGCNA analysis, four cancer progression and metastasis related modules were detected. After PPI and survival analysis, we identified and validated
HEY2 as a hub gene, high expression and poor survival at the metastatic stage. Functionally, berberine inhibited the survival, invasion and migration of CRC cells in vitro and in vivo. Mechanistically, berberine treatment down-regulated the expression of
HEY2, metastasis related protein E-cadherin, β-catenin and Cyclin D1 during Mesenchymal epithelial transformation (MET). Berberine and
HEY2 showed a significant interaction, and berberine binded to
HEY2 protein at the residue HIS-99 interface with a hydrogen-bond distance of 1.9A.
CONCLUSIONS: We revealed that berberine could significantly inhibit the expression of hub gene HEY2 and metastasis related proteins E-cadherin and β-catenin and Cyclin D1 during MET in CRC lung and liver metastases. In total,
HEY2 was a promising candidate biomarker for prognosis and molecular characteristics in CRC metastasis.