Han Xiu-Guo, Li Yan, Mo Hui-Min, Li Kang, Lin Du, Zhao Chang-Qing, Zhao Jie, Tang Ting-Ting
Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Zhizaoju Road 639, Shanghai, 200011, China.
Institution of Hematology, Xuzhou Medical College, Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Jiangsu Province, China.
Tumour Biol. 2016 Jul;37(7):8857-67. doi: 10.1007/s13277-015-4757-4. Epub 2016 Jan 9.
Tissue inhibitors of metalloproteinases (TIMPs) inhibit matrix metalloproteinases (MMPs) to limit degradation of the extracellular matrix. Low levels of TIMP3 have been demonstrated in cancer tissues at advanced clinical stages, with positive distant metastasis and chemotherapeutic resistance. We examined the role of TIMP3 in osteosarcoma (OS) cell invasiveness and chemoresistance. TIMP3 was overexpressed or knocked down in the human OS cell lines Saos2 and MG63. Cell migration and invasion capacities were then evaluated using Transwell assays, and resistance to cisplatin was assessed by CCK-8 assay and flow cytometry. Real-time PCR and western blotting were used to investigate activation of signaling pathways downstream of TIMP3. Overexpression of TIMP3 inhibited the migration and invasion of Saos2 and MG63 cells, while knockdown of TIMP3 had the opposite effect. Cell survival after exposure to cisplatin was inhibited by TIMP3 overexpression in both Saos2 and MG63 cells. Consistently, downregulation of TIMP3 gene expression significantly decreased the sensitivity of OS cells to cisplatin treatment. MMP1, MMP2, Bcl-2, and Akt1 were all downregulated following TIMP3 overexpression, while Bax and cleaved caspase-3 were upregulated. TIMP3 knockdown had opposite effects on the regulation of these genes. Taken together, our findings suggest TIMP3 as a new target for inhibition of OS progression and chemotherapeutic resistance.
金属蛋白酶组织抑制剂（TIMPs）可抑制基质金属蛋白酶（MMPs），从而限制细胞外基质的降解。在临床晚期、存在远处转移且具有化疗耐药性的癌症组织中，已证实TIMP3水平较低。我们研究了TIMP3在骨肉瘤（OS）细胞侵袭和化疗耐药中的作用。在人OS细胞系Saos2和MG63中过表达或敲低TIMP3。然后使用Transwell实验评估细胞迁移和侵袭能力，并通过CCK-8实验和流式细胞术评估对顺铂的耐药性。采用实时PCR和蛋白质印迹法研究TIMP3下游信号通路的激活情况。TIMP3的过表达抑制了Saos2和MG63细胞的迁移和侵袭，而TIMP3的敲低则产生相反的效果。在Saos2和MG63细胞中，TIMP3的过表达均抑制了顺铂处理后的细胞存活。同样，TIMP3基因表达的下调显著降低了OS细胞对顺铂治疗的敏感性。TIMP3过表达后，MMP1、MMP2、Bcl-2和Akt1均下调，而Bax和裂解的caspase-3上调。TIMP3敲低对这些基因的调控产生相反的影响。综上所述，我们的研究结果表明TIMP3是抑制OS进展和化疗耐药的新靶点。
