甲状旁腺素通过调控初级纤毛的表达促进软骨肉瘤细胞增殖和侵袭

摘要

目的 探究甲状旁腺素(parathyroid hormone,PTH)对软骨肉瘤细胞增殖和侵袭能力的影响,及其与初始纤毛表达调控之间的相互关系.方法 应用不同浓度PTH刺激软骨肉瘤细胞SW1353,缺氧诱导纤毛表达和水合氯醛破坏纤毛结构后,通过CCK8细胞活性实验,Western blot及Transwell实验检测SW1353细胞的增殖和侵袭能力,免疫荧光计数初始纤毛表达情况,荧光定量PCR检测相关目的基因GLI1、PTCH1和IFT88的表达情况.结果 PTH能促进软骨肉瘤细胞增殖,且呈浓度依赖性,其促增殖效应与纤毛结构的完整性有关;PTH能上调SW1353细胞的侵袭能力,增加基质金属蛋白酶MMP9的表达,破坏纤毛结构后,PTH促进软骨肉瘤的侵袭效应受到抑制;PTH下调SW1353细胞初始纤毛的数量,其效应与纤毛结构完整性有关;PTH能调控Hedgehog通路靶基因GLI1、PTCH1及纤毛功能相关基因IFT88的表达.结论 PTH能促进软骨肉瘤细胞的增殖和侵袭,同时抑制软骨肉瘤细胞初级纤毛的表达并影响其下游靶基因表达,PTH可能通过作用于初始纤毛来调控软骨肉瘤的恶性生物学特性.%Objective To investigate the effect of parathyroid hormone(PTH)on the proliferation and invasion of chondrosarcoma cells,and the relationship between PTH and the regulation of primary cilia expression.Methods After stimulation of the chondrosarcoma cell line SW1353 with different concentrations of PTH,induction of the expression of cilia with hypoxia and destruction of cilia structure with chloral hydrate,the cell viability was detected by CCK8 assay,the proliferation and invasion of SW1353 by Western blotting and Transwell method,the primary cilia expression by immunofluorescence assay and the GLI1,PTCH1 and IFT88 expression levels by real-time PCR.Results PTH could promote the proliferation of chondrosarcoma cells in a concentration-dependent manner and this effect was correlated with the structural integrity of the primary cilia.PTH could up-regulate the invasive ability of SW1353 cells and increase the expression levels of MMP9,which was suppressed when the primary cilia structure was damaged.Additionally,it was found that PTH could down-regulate the number of primary cilia,which was related to the structural integrity of the primary cilia.It could also regulate the expression levels of GLI1 and PTCH1,the target genes in Hedgehog pathway,and the expression levels of IFT88,the gene associated with the cilia function.Conclusion PTH can promote the proliferation and invasion of chondrosarcoma cells,down-regulate the expression of primary cilia and the downstream target genes.PTH may regulate the malignant biological features of chondrosarcoma by regulating the primary cilia expression.