甲基-β-环糊精通过限制脂质代谢抑制横纹肌肉瘤细胞的增殖、迁移和侵袭

Methyl-β-cyclodextrin inhibits the proliferation,migration and invasion of rhabdomyosarcoma cells by restricting lipid metabolism

目的探讨抑制脂质合成对人骨骼横纹肌肉瘤细胞的影响及其分子机制。方法采用肿瘤基因表达数据库检查人骨骼肌细胞(HSMC)和人骨骼横纹肌肉瘤细胞脂质合成相关基因固醇调节元件结合蛋白1(SREBP1)和角鲨烯环氧化酶(SQLE)mRNA表达水平并通过qRT-PCR验证。通过细胞增殖实验选出脂质抑制剂甲基-β-环糊精(mβCD)的细胞处理浓度。分别将3株人骨骼横纹肌肉瘤细胞(RD、SJCRH30和A673)作为对照组,1 mmol/L mβCD处理的三株人骨骼横纹肌肉瘤细胞分别作为实验组。平板克隆实验、软琼脂集落形成实验、细胞迁移及侵袭实验和裸鼠成瘤实验检测对照组和mβCD处理组人骨骼横纹肌肉瘤细胞增殖、迁移、侵袭和肿瘤生长的变化。通过脂质组学及三酰甘油(TG)分析探究mβCD抑制人骨骼横纹肌肉瘤细胞恶性的分子机制。结果相对于HSMC,人骨骼横纹肌肉瘤细胞脂质合成相关基因SREBP1、SQLE表达增高(P<0.001);与对照组相比,mβCD组人骨骼横纹肌肉瘤细胞的增殖、平板克隆、迁移侵袭和集落形成能力减弱(P<0.05);裸鼠肿瘤体积和肿瘤质量增加也减少(P<0.05);脂质组学和TG试剂盒检测结果发现,与对照组相比,mβCD组TG含量降低(P<0.01)。结论mβCD可能通过降低TG等脂质代谢过程从而抑制人骨骼横纹肌肉瘤细胞的增殖、迁移和侵袭等恶性生物学行为。

Objective To investigate the effect of inhibiting lipid synthesis on human skeletal rhabdomyosarcoma cells and its molecular mechanism.Methods The mRNA expression levels of lipid synthesis related genes sterol regulatory element binding protein 1(SREBP1)and squalene cyclooxygenase(SQLE)in human skeletal muscle cells(HSMC)and human skeletal rhabdomyosarcoma cells were detected by tumor gene expression database and verified by qRT-PCR.The concentration of methyl-β-cyclodextrin(mβCD)was determined by cell proliferation assay.The control group consisted of three human skeletal rhabdomyosarcoma cell lines(RD,SJCRH30,A673),while the experimental group comprised three human skeletal rhabdomyosarcoma cell lines treated with 1mmol/L mβCD.Plate clone formation assay,soft agar,colony formation assay,cell migration assay and cell invasion assay,and tumor formation in nude mice employed to assess changes in proliferation,migration,invasion,and tumor growth of human skeletal rhabdomyosarcoma cells between the control group and mβCD treatment group.The molecular mechanism of mβCD inhibiting the malignancy of human skeletal rhabdomyosarcoma cells was explored by lipoomics and triglyceride(TG)detection.Results Compared with HSMC,the expressions of lipid synthesis related genes SREBP1 and SQLE significantly increased in human skeleton rhabdomyosarcoma cells(P<0.001).Compared with the control group,the proliferation,plate cloning,migration,invasion and colony formation ability of human skeleton rhabdomyosarcoma cells in the mβCD group significantly decreased(P<0.05).The growth of tumor volume and weight in nude mice was also significantly reduced(P<0.05).The lipidomics results and TG kit analysis revealed a significant reduction in TG content in the mβCD group compared to the control group(P<0.01).Conclusion mβCD may inhibit the malignant biological behaviors such as proliferation,migration and invasion of human skeleton rhabdomyosarcoma cells by reducing TG and other lipid metabolism.