title: Systemic distribution, subcellular localization and differential expression of sphingosine-1-phosphate receptors in benign and malignant human tissues.
abstract: OBJECTIVE
  Five sphingosine-1-phosphate receptors (S1PR): S1PR1, S1PR2, S1PR3, S1PR4 and S1PR5 (S1PR1-5) have been shown to be involved in the proliferation and progression of various cancers. However, none of the S1PRs have been systemically investigated. In this study, we performed immunohistochemistry (IHC) for S1PR1-S1PR5 on different tissues, in order to simultaneously determine the systemic distribution, subcellular localization and expression level of all five S1PRs.
  
  
  METHODS
  We constructed tissue microarrays (TMAs) from 384 formalin-fixed paraffin-embedded (FFPE) blocks containing 183 benign and 201 malignant tissues from 34 human organs/systems. Then we performed IHC for all five S1PRs simultaneously on these TMA slides. The distribution, subcellular localization and expression of each S1PR were determined for each tissue. The data in benign and malignant tissues from the same organ/tissue were then compared using the Student's t-test. In order to reconfirm the subcellular localization of each S1PR as determined by IHC, immunocytochemistry (ICC) was performed on several malignant cell lines.
  
  
  RESULTS
  We found that all five S1PRs are widely distributed in multiple human organs/systems. All S1PRs are expressed in both the cytoplasm and nucleus, except S1PR3, whose IHC signals are only seen in the nucleus. Interestingly, the S1PRs are rarely expressed on cellular membranes. Each S1PR is unique in its organ distribution, subcellular localization and expression level in benign and malignant tissues. Among the five S1PRs, S1PR5 has the highest expression level (in either the nucleus or cytoplasm), with S1PR1, 3, 2 and 4 following in descending order. Strong nuclear expression was seen for S1PR1, S1PR3 and S1PR5, whereas S1PR2 and S1PR4 show only weak staining. Four organs/tissues (adrenal gland, liver, brain and colon) show significant differences in IHC scores for the multiple S1PRs (nuclear and/or cytoplasmic), nine (stomach, lymphoid tissues, lung, ovary, cervix, pancreas, skin, soft tissues and uterus) show differences for only one S1PR (cytoplasmic or nuclear), and twenty three organs/tissues show no significant difference in IHC scores for any S1PR (cytoplasmic or nuclear) between benign and malignant changes.
  
  
  CONCLUSIONS
  This is the first study to evaluate the expression level of all S1PRs in benign and malignant tissues from multiple human organs. This study provides data regarding the systemic distribution, subcellular localization and differences in expression of all five S1PRs in benign and malignant changes for each organ/tissue.
authors.affiliation: Department of Gastrointestinal Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China; Department of Pathology, University of Mississippi Medical Center, MS 39216, USA. Electronic address: cwang@umc.edu.
authors.forename: Chunyi
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Biology, Tougaloo College, Tougaloo, MS 39174, USA. Electronic address: jmao@tougaloo.edu.
authors.forename: Jinghe
authors.initials: J
authors.lastname: Mao
authors.affiliation: Department of Pathology, University of Mississippi Medical Center, MS 39216, USA. Electronic address: SDRedfield@umc.edu.
authors.forename: Samantha
authors.initials: S
authors.lastname: Redfield
authors.affiliation: Cancer Institute, University of Mississippi Medical Center, MS 39216, USA. Electronic address: ymo@umc.edu.
authors.forename: Yinyuan
authors.initials: Y
authors.lastname: Mo
authors.affiliation: Department of Pathology, University of Mississippi Medical Center, MS 39216, USA. Electronic address: jlage@umc.edu.
authors.forename: Janice M
authors.initials: JM
authors.lastname: Lage
authors.affiliation: Department of Pathology, University of Mississippi Medical Center, MS 39216, USA; Cancer Institute, University of Mississippi Medical Center, MS 39216, USA. Electronic address: xzhou@umc.edu.
authors.forename: Xinchun
authors.initials: X
authors.lastname: Zhou
mesh_terms: D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D009369:Neoplasms / Q000378:metabolism*; D009928:Organ Specificity; D021381:Protein Transport; D049349:Receptors, Lysosphingolipid / Q000235:genetics / Q000378:metabolism*
chemical_list: D049349:Receptors, Lysosphingolipid
keywords: Adrenal gland; Cancer; Colon; Immunohistochemistry (IHC); Liver; Lung; Neural system; Ovary; Sphingosine-1-phosphate (S1P) receptors (S1PRs); Stomach
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