Transcriptional profile in SK-LM_S1 cells mediated by abrogation of MAPK7 and VCAN gene expression [Part 1]

The ERK5 signaling pathway has emerged as a novel key player in soft tissue sarcoma biology, mediating tumour growth and progression. Here we show how the expression of VCAN, a member of the chondroitin sulphate proteoglycan family, is modulated by the ERK5 pathway at the transcriptional level and mediates biological processes such as migration, adhesion, proliferation and tumourigenesis in vivo. This conclusion is based on genetic (shRNA, overexpression) and pharmacological approaches using the human soft tissue sarcoma cell lline Sk-LM-S1 a. In particular, VCAN knockdown mimics ERK5 knockdown in vitro and in vivo, whereas VCAN overexpression restores the phenotypes observed after ERK5 knockdown. In addition, transcriptomic studies show that VCAN is a central mediator in the ERK5-associated transcriptional landscape, supporting its role in diverse biological processes. Taken together, these data support VCAN as a new key target in the ERK5 pathway. Overall design: Following effective knockdown of VCAN and MAPK7 gene expression in SK-LM-S1 cells using specific shRNA, samples were collected and transcriptomic profile analyzed by RNA-seq.

ERK5信号通路（ERK5 signaling pathway）已被证实为软组织肉瘤（soft tissue sarcoma）发生发展中的新型关键调控因子，介导肿瘤的生长与进展过程。本研究揭示了软骨素硫酸盐蛋白多糖（chondroitin sulphate proteoglycan）家族成员VCAN的表达如何在转录水平受ERK5通路调控，并在体内介导细胞迁移、黏附、增殖及致瘤等多种生物学过程。本研究结论基于以人软组织肉瘤细胞系Sk-LM-S1为模型的遗传干预手段（包括shRNA、过表达（overexpression））与药理学方法。具体而言，体外与体内实验中，VCAN敲低可模拟ERK5敲低的细胞表型，而VCAN过表达则可恢复ERK5敲低后出现的表型。此外，转录组学研究显示，VCAN是ERK5相关转录组景观中的核心介导因子，佐证了其在多种生物学过程中的作用。综上，上述数据支持VCAN作为ERK5通路中新的关键靶点。总体实验设计：利用特异性shRNA对SK-LM-S1细胞中的VCAN与MAPK7基因进行有效敲低后，收集样本并通过RNA测序（RNA-seq）分析其转录组特征。