Zhou Qiwen, You Yang, Zhao Yingying, Xiao Shuxiu, Song Zhengqing, Huang Chuxin, Qian Jiali, Lu Weiqi, Tong Hanxing, Zhang Yong, Wang Zhiming, Li Wei, Zhang Chenlu, Guo Xi, Luo Rongkui, Hou Yingyong, Cui Jiefeng, Lu Lili, Zhou Yuhong
Department of Oncology, Zhongshan Hospital, Fudan University, Shanghai, 200032, China.
Liver Cancer Institute, Zhongshan Hospital, Fudan University and Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai, 200032, China.
Cell Oncol (Dordr). 2025 Apr;48(2):455-470. doi: 10.1007/s13402-024-01008-7. Epub 2024 Nov 29.
Leiomyosarcoma (LMS) is an aggressive mesenchymal malignant tumor with poor therapeutic options, but the molecular mechanisms underlying LMS remain largely unknown. Increasing evidence indicates that transient receptor potential vanilloid 4 (TRPV4) levels are closely related to the advancement of various malignant tumors through diverse molecular mechanisms. However, the roles and regulatory mechanisms of TRPV4 in LMS progression remain unclear.
Immunohistochemistry, Western blot, and immunofluorescence were used to investigate the relationship between TRPV4 expression and LMS. Survival analysis was conducted to evaluate the association between TRPV4 levels and prognosis in LMS patients. Intracellular Ca measurement, colony formation, CCK-8, wound healing and Transwell assays and peritoneal metastasis mouse model were used to verify the effect of TRPV4 activity and expression on LMS proliferation and metastasis. RNA-seq and proteomics were performed to explore the underlying mechanism.
TRPV4 was upregulated in LMS tissues and cells and served as a novel prognostic factor. Moreover, TRPV4 overexpression enhanced cell proliferation, cell migration and invasion of LMS cells in vitro, as well as promoted tumor metastasis in vivo, which could be blocked by HC067047 intervention or TRPV4 knockdown. Combined RNA-seq and proteomics analysis of KEGG pathway indicated that ECM receptor interaction was obviously activated. Extracellular matrix protein 1 (ECM1) was identified as downstream gene of TRPV4. Mechanistically, TRPV4 overexpression increased ECM1 level and activated the FAK/PI3K/AKT/GSK3β pathway, which could be reversed by TRPV4 knockdown or LY294002 treatment. Moreover, ECM1 overexpression enhanced the activation of FAK/PI3K/AKT/GSK3β pathway. And simultaneous overexpression of TRPV4 and ECM1 synergistically activated this pathway.
Our findings provide a novel mechanism by which TRPV4 directly activates Ca/FAK/PI3K/AKT/GSK3β pathway and further indirectly enhances the FAK/PI3K/AKT/GSK3β pathway through the promotion and secretion of ECM1 to promote LMS malignant progression. Targeting the TRPV4/FAK axis might be a promising potential strategy for prognosis and treatment of LMS.
平滑肌肉肉瘤（LMS）是一种侵袭性间充质恶性肿瘤，治疗选择有限，但其潜在分子机制仍在很大程度上未知。越来越多的证据表明，瞬时受体电位香草酸受体4（TRPV4）水平通过多种分子机制与各种恶性肿瘤的进展密切相关。然而，TRPV4在LMS进展中的作用和调控机制仍不清楚。
采用免疫组织化学、蛋白质免疫印迹和免疫荧光法研究TRPV4表达与LMS之间的关系。进行生存分析以评估LMS患者中TRPV4水平与预后的相关性。采用细胞内钙测量、集落形成、CCK-8、伤口愈合和Transwell实验以及腹膜转移小鼠模型来验证TRPV4活性和表达对LMS增殖和转移的影响。进行RNA测序和蛋白质组学分析以探索潜在机制。
TRPV4在LMS组织和细胞中上调，并作为一种新的预后因素。此外，TRPV4过表达增强了LMS细胞在体外的增殖、迁移和侵袭能力，以及在体内促进肿瘤转移，这可被HC067047干预或TRPV4基因敲低所阻断。KEGG通路的联合RNA测序和蛋白质组学分析表明，细胞外基质受体相互作用明显被激活。细胞外基质蛋白1（ECM1）被鉴定为TRPV4的下游基因。机制上，TRPV4过表达增加了ECM1水平并激活了FAK/PI3K/AKT/GSK3β通路，这可通过TRPV4基因敲低或LY294002处理来逆转。此外，ECM1过表达增强了FAK/PI3K/AKT/GSK3β通路的激活。并且TRPV4和ECM1的同时过表达协同激活了该通路。
我们的研究结果提供了一种新机制，即TRPV4直接激活Ca/FAK/PI3K/AKT/GSK3β通路，并通过促进和分泌ECM1进一步间接增强FAK/PI3K/AKT/GSK3β通路，从而促进LMS的恶性进展。靶向TRPV4/FAK轴可能是LMS预后和治疗的一种有前景的潜在策略。
