Data Sheet 1_SOX2 induces LPCAT1 expression to promote cholesterol metabolic reprogramming-mediated invasion and metastasis in osteosarcoma.zip

BackgroundSOX2 and LPCAT1 are implicated in tumor progression, but their roles in osteosarcoma pathogenesis and cholesterol metabolism remain unclear.
MethodSOX2 and LPCAT1 expression in osteosarcoma tissues and cell lines was assessed via qRT-PCR and Western blot. Functional assays (CCK-8, wound healing and Transwell) evaluated proliferation, migration, and invasion of osteosarcoma cells. SOX2-LPCAT1 binding was confirmed by dual-luciferase reporter assay and ChIP assays. RNA sequencing and bioinformatics analyses explored cholesterol metabolism pathways. In vitro and in vivo models (xenograft tumor model and lung metastasis model) validated mechanistic roles.
ResultSOX2 and LPCAT1 were overexpressed in osteosarcoma. LPCAT1 or SOX2 overexpression promoted malignant behaviors and cholesterol metabolism (free cholesterol/total cholesterol levels, SREBP1/INSIG1 expression) of osteosarcoma cells, while shSOX2 or shLPCAT1 did the opposite. SOX2 transcriptionally activated LPCAT1. LPCAT1 reversed shSOX2-induced suppression, while LPCAT1 knockdown attenuated SOX2-driven oncogenicity. In vivo, LPCAT1 enhanced tumor growth, lung metastasis, and cholesterol metabolism, while these effects were counteracted by SOX2 inhibition.
ConclusionThe SOX2/LPCAT1 axis drives osteosarcoma progression by modulating cholesterol metabolism.

背景：SOX2与LPCAT1已被证实与肿瘤进展相关，但其在骨肉瘤发生发展及胆固醇代谢中的作用仍未明确。方法：本研究通过实时荧光定量PCR（qRT-PCR）与蛋白质印迹法（Western blot）检测了骨肉瘤组织及细胞系中SOX2与LPCAT1的表达水平；采用CCK-8实验、划痕愈合实验及Transwell实验评估骨肉瘤细胞的增殖、迁移与侵袭能力；通过双荧光素酶报告基因实验与染色质免疫沉淀（ChIP）实验验证了SOX2与LPCAT1的结合关系；借助RNA测序与生物信息学分析探究了胆固醇代谢相关通路；并通过体外实验与体内实验（异种移植瘤模型及肺转移模型）验证了其机制作用。结果：SOX2与LPCAT1在骨肉瘤中呈高表达状态。过表达LPCAT1或SOX2可促进骨肉瘤细胞的恶性生物学行为及胆固醇代谢进程，包括游离胆固醇/总胆固醇水平、SREBP1与INSIG1的表达量，而shSOX2或shLPCAT1则会产生相反效果。SOX2可通过转录激活方式调控LPCAT1的表达。LPCAT1可逆转shSOX2诱导的细胞增殖抑制效应，而敲低LPCAT1则会削弱SOX2介导的肿瘤恶性表型。体内实验结果显示，LPCAT1可促进肿瘤生长与肺转移，并增强胆固醇代谢水平，而抑制SOX2可抵消上述效应。结论：SOX2/LPCAT1信号轴通过调控胆固醇代谢进程促进骨肉瘤进展。