桑根酮C通过促进β-catenin的泛素化水平抑制胶质母细胞瘤的迁移侵袭能力

Sanggenon C inhibits glioblastoma cell migration and invasion by promoting ubiquitination of β-catenin

摘要:

胶质母细胞瘤属于浸润性的恶性肿瘤，目前其化疗新药物的寻找和治疗仍待突破。桑根酮C (Sanggenon C, SC)来源于桑白皮，在多种癌症中发挥着抗肿瘤功效。本研究利用显微镜拍摄、transwell实验和免疫荧光实验验证了SC对胶质母细胞瘤的迁移侵袭能力的影响；基因富集、实时荧光定量PCR (real-time qPCR)以及泛素化实验用于阐明SC抑制胶质母细胞瘤迁移侵袭能力的分子机制。显微镜拍摄结果显示，对胶质母细胞瘤进行加药处理后细胞形态明显回缩，细胞迁移侵袭能力被削弱；Transwell实验和免疫荧光实验也验证了SC能抑制胶质母细胞瘤迁移侵袭能力的猜测；基因富集实验结果表明SC可能调控迁移侵袭相关基因的表达，并且影响Wnt/β-catenin信号通路的活性；Western blotting揭示了SC可调控β-catenin的泛素化水平，并且抑制β-catenin及其下游蛋白的表达；Real-time qPCR实验结果在转录水平上佐证Western blotting的结果。SC通过调控β-catenin的泛素化水平抑制胶质母细胞瘤的迁移侵袭能力，为胶质母细胞瘤的治疗提供了新的思路。

关键词:胶质母细胞瘤;桑根酮C;迁移;侵袭;β-catenin泛素化

Abstract:

Glioblastoma is a malignant and highly invasive tumor, which requires new approaches to search for chemotherapeutic agents. Sanggenon C (SC) mainly exists in the root bark of white mulberry. Although its anti-tumor effects have been reported in some cancers, the mechanism remains unclear. In this study, we used microscopic observation, transwell assay, and immunofluorescence assay to verify the effect of Sanggenon C on the migration and invasion of glioblastoma cells. We then carried out the gene set enrichment analysis (GESA), real-time qPCR assay and ubiquitination assay to delineate the molecule mechanism by which Sanggenon C affects the migration and invasion ability of glioblastoma. With the addition of Sanggenon C, glioblastoma cells were rounded up, with the migration and invasion ability weakened as verified by transwell assay and immunofluorescence assay. The results of GESA suggested that SC might regulate the expression of genes associated with migration and invasion and affect the activity of Wnt/β-catenin signaling pathway. Western blotting revealed that Sanggenon C promoted the ubiquitination of β-catenin to reduce the levels of β-catenin and its downstream proteins. This was further supported by the results of real-time qPCR analysis of target genes of β-catenin. Taken together, SC inhibits glioblastoma cell migration and invasion by enhancing β-catenin ubiquitination. Our work suggests a new direction for the treatment of glioblastoma.

Key words:glioblastoma;Sanggenon C;migration;invasion;the ubiquitination of β-catenin