title: TIMP3 regulates osteosarcoma cell migration, invasion, and chemotherapeutic resistances.
abstract: 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.
authors.affiliation: 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.
authors.forename: Xiu-Guo
authors.initials: XG
authors.lastname: Han
authors.affiliation: 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.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Institution of Hematology, Xuzhou Medical College, Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Jiangsu Province, China.
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: Mo
authors.affiliation: 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.
authors.forename: Kang
authors.initials: K
authors.lastname: Li
authors.affiliation: Department of Orthopedic Surgery, Shanghai First People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Du
authors.initials: D
authors.lastname: Lin
authors.affiliation: 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.
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Zhao
authors.affiliation: 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. medzhaojie@163.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhao
authors.affiliation: 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. tingtingtang@hotmail.com.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Tang
mesh_terms: D053148:Caspase 3 / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D002470:Cell Survival / Q000235:genetics; D002945:Cisplatin / Q000494:pharmacology; D015536:Down-Regulation / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000235:genetics; D020778:Matrix Metalloproteinase 2 / Q000235:genetics; D009361:Neoplasm Invasiveness / Q000235:genetics* / Q000473:pathology; D012516:Osteosarcoma / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics*; D015854:Up-Regulation / Q000235:genetics; D051028:bcl-2-Associated X Protein / Q000235:genetics
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; C088390:TIMP3 protein, human; D019717:Tissue Inhibitor of Metalloproteinase-3; D051028:bcl-2-Associated X Protein; C494918:AKT1 protein, human; D051057:Proto-Oncogene Proteins c-akt; D053148:Caspase 3; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1; D002945:Cisplatin
keywords: Cisplatin sensitivity; Invasion; Migration; Osteosarcoma; TIMP3