title: Novel esophageal squamous cell carcinoma bone metastatic clone isolated by scintigraphy, X ray and micro PET/CT.
abstract: OBJECTIVE
  To establish a Chinese esophageal squamous cell carcinoma (ESCC) cell line with high bone metastasis potency using (99m)Tc-methylene diphosphonate ((99m)Tc-MDP) micro-pinhole scintigraphy, X ray and micro-positron emission tomography/computed tomography (PET/CT) for exploring the mechanism of occurrence and development in esophageal cancer.
  
  
  METHODS
  The cells came from a BALB/c nu/nu immunodeficient mouse, and oncogenic tumor tissue was from a surgical specimen from a 61-year-old male patient with ESCC. The cell growth curve was mapped and analysis of chromosome karyotype was performed. Approximately 1 × 10⁶ oncogenic cells were injected into the left cardiac ventricle of immunodeficient mice. The bone metastatic lesions of tumor-bearing mice were detected by (99m)Tc-MDP scintigraphy, micro-PET/CT and X-ray, and were resected from the mice under deep anesthesia. The bone metastatic cells in the lesions were used for culture and for repeated intracardiac inoculation. This in vivo/in vitro experimental metastasis study was repeated for four cycles. All of the suspicious bone sites were confirmed by pathology. Real-time polymerase chain reaction was used to compare the gene expression in the parental cells and in the bone metastatic clone.
  
  
  RESULTS
  The surgical specimen was implanted subcutaneously in immunodeficient mice and the tumorigenesis rate was 100%. First-passage oncogenic cells were named CEK-Sq-1. The chromosome karyotype analysis of the cell line was hypotriploid. The bone metastasis rate went from 20% with the first-passage oncogenic cells via intracardiac inoculation to 90% after four cycles. The established bone metastasis clone named CEK-Sq-1BM had a high potential to metastasize in bone, including mandible, humerus, thoracic and lumbar vertebrae, scapula and femur. The bone metastasis lesions were successfully detected by micro-pinhole bone scintigraphy, micro-PET/CT, and X-ray. The sensitivity, specificity and accuracy of the micro-pinhole scintigraphy, X-ray, and micro-PET/CT imaging examinations were: 89.66%/32%/80%, 88.2%/100%/89.2%, and 88.75%/77.5%/87.5%, respectively. Some gene expression difference was found between parental and bone metastasis cells.
  
  
  CONCLUSIONS
  This newly established Chinese ESCC cell line and animal model may provide a useful tool for the study of the pathogenesis and development of esophageal carcinoma.
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Bi-Zeng
authors.initials: BZ
authors.lastname: Zhao
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Cao
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Jin-Chen
authors.initials: JC
authors.lastname: Shao
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Yan-Bing
authors.initials: YB
authors.lastname: Sun
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Li-Min
authors.initials: LM
authors.lastname: Fan
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Chun-Yu
authors.initials: CY
authors.lastname: Wu
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Liang
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Bao-Feng
authors.initials: BF
authors.lastname: Guo
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Guang
authors.initials: G
authors.lastname: Yang
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Wen-Hui
authors.initials: WH
authors.lastname: Xie
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Qing-Cheng
authors.initials: QC
authors.lastname: Yang
authors.affiliation: Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China.
authors.forename: Shun-Fang
authors.initials: SF
authors.lastname: Yang
mesh_terms: D000818:Animals; D001859:Bone Neoplasms / Q000000981:diagnostic imaging / Q000235:genetics / Q000556:secondary*; D002294:Carcinoma, Squamous Cell / Q000000981:diagnostic imaging / Q000235:genetics / Q000556:secondary* / Q000601:surgery; D045744:Cell Line, Tumor; D002877:Chromosomes, Human; D004938:Esophageal Neoplasms / Q000235:genetics / Q000473:pathology* / Q000601:surgery; D000077277:Esophageal Squamous Cell Carcinoma; D015972:Gene Expression Regulation, Neoplastic; D064593:Heterografts; D006801:Humans; D007621:Karyotyping; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D064847:Multimodal Imaging / Q000379:methods*; D009368:Neoplasm Transplantation; D049268:Positron-Emission Tomography; D011237:Predictive Value of Tests; D019275:Radiopharmaceuticals; D013669:Technetium Tc 99m Medronate; D055114:X-Ray Microtomography
chemical_list: D019275:Radiopharmaceuticals; D013669:Technetium Tc 99m Medronate
keywords: Bone metastasis; Cell line; Esophageal squamous cell carcinoma; Molecular imaging; Real-time polymerase chain reaction