title: [Inoculation of the cells transfected with IP10 inhibited experimental tumor metastases in vivo].
abstract: OBJECTIVE
  To explore the inhibitory effects of IP10 on the experimental tumor metastases of a mammary carcinoma cell line 4T1 in vivo.
  
  
  METHODS
  4T1 cells were transfected with pcDNA3-IP10 plasmid and the positive clones (IP10-4T1) were screened in the presence of G418. The parental 4T1 cells and 4T1 cells transfected with pcDNA3 (pcDNA3-4T1) were used as controls. The expression of IP10 mRNA was examined with RT-PCR. The chemotactic activity of the cell-cultured supernatant for the activated lymphocytes was assessed with chemotaxis assay. All BALB/c mice were divided into 3 equal groups: IP10-4T1, parental 4T1 and pcDNA3-4T1 (6 mice in each group). Seven days after mice were inoculated with 2 x 10(6) 4T1, pcDNA3-4T1 or IP10-4T1 cells, 1 x 10(5) 4T1 cells were injected into mice in tail vein. On day 14 tumors were sectioned. On day 28 mice were sacrificed and the lung weight, metastatic forci on the lung surface, disseminated metastases in the lungs and the number of clonogenic metastases of 4T1 cells were observed. The splenocytes were isolated from tumor-bearing mice, and the cytotoxicity of the splenocytes was evaluated by CFSE/7-AAD method.
  
  
  RESULTS
  The transcription of IP10 mRNA increased in IP10-4T1 cells compared to parental 4T1 and pcDNA3-4T1 cells (P = 0.002). Moreover, accumulated lymphocytes migrated to the supernatants of IP10-4T1 cells, which can be abrogated by anti-CXCR3 (P < 0.05). Compared to controls, inoculation of IP10-4T1 cells resulted in the decreased disseminated metastases as indicated by dramatically reduced lung metastatic forci on the surface of the lungs; the lung weight was lighter (IP10-4T1, 0.27 +/- 0.02 g v.s. 4T1, 0.48 +/- 0.08 g and pcDNA3-4T1, 0.43 +/- 0.16 g, P = 0.021); the number of clonogenic metastatic 4T1 cells enumerated in ten fields decreased greatly (IP10-4T1, 2.6 +/- 1.7 v.s. 4T1, 34 +/- 6.3 and pcDNA3-4T1, 33 +/- 2.3, P < 0.05); histological assay showed that metastasis was not found in the lungs of the 4/6 of mice in IP10-4T1 group, and was seen in all the mice of parental 4T1 and pcDNA3-4T1 groups. The splenocytes from the mice inoculated with IP10-4T1 cells exhibited stronger cytotoxic activity against 4T1 target cells at different ratios of effector versus target cells (3:1, 9:1, 27:1) than controls (P < 0.05).
  
  
  CONCLUSIONS
  IP10 expressed locally could inhibit disseminated metastasis of circulating 4T1 tumor cells by enhancing anti-tumor cellular immune responses.
authors.affiliation: Department of Immunology, Shanghai Medical College, Fudan University, Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai 200032, China.
authors.forename: Xiu-li
authors.initials: XL
authors.lastname: Yang
authors.forename: Yi-wei
authors.initials: YW
authors.lastname: Chu
authors.forename: Lin
authors.initials: L
authors.lastname: Xu
authors.forename: Bao-hua
authors.initials: BH
authors.lastname: Li
authors.forename: Si-dong
authors.initials: SD
authors.lastname: Xiong
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D054357:Chemokine CXCL10 / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D002633:Chemotaxis / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D008175:Lung Neoplasms / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008297:Male; D008325:Mammary Neoplasms, Experimental / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009368:Neoplasm Transplantation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D054357:Chemokine CXCL10; D017077:Culture Media, Conditioned; D012333:RNA, Messenger