title: [Influence of receptor activity modifying protein 1 overexpression on enhancing effect of calcitonin gene-related peptide on MG-63 cells proliferation].
abstract: OBJECTIVE
  To investigate the influnce of receptor activity modifying protein 1(RAMP-1) overexpression on enhancing effect of calcitonin gene-related peptide on MG-63 cells proliferation.
  
  
  METHODS
  Cultured MG-63 osteoblasts in exponential phase of growth were randomly divided into three groups: RAMP-1 overexpression group, empty vector control group and negative control group. RAMP-1 eukaryotic expression vector was constructed and stably transfected into MG-63 cells. Realtime-polymerase chain reaction, Western blotting and immunofluroescence were used respectively to detect the expression of calcitonin receptor-like receptor (CRLR) in the cells and its distribution on cell membrane. The status of proliferation was detected respectively at 0, 24, 48, 72, 96 h by cell counting kit-8 (CCK-8) and cells were collected to analyze their cycle respectively at 0, 8, 16, 24 h by flow cytometry.
  
  
  RESULTS
  CRLR protein and mRNA expression levels of MG-63 cells in RAMP-1 overexpression group were significantly higher than the other two groups (P < 0.05). The A value of RAMP-1 overexpression group at 24, 48, 72, 96 h were 0.628 ± 0.175, 0.896 ± 0.592, 1.055 ± 0.004, 1.179 ± 0.618, respectively, which were significantly higher than that of the other two groups (P < 0.05). The difference was most pronounced at 72 h. S-phase fraction of RAMP-1 overexpression group was (1.25 ± 0.13)%, (68.79 ± 0.56)%, (64.49 ± 1.59)%, (57.82 ± 0.75)%, respectively, which were significantly higher than the other two groups (P < 0.05). The difference was most pronounced at 8 h.
  
  
  CONCLUSIONS
  RAMP-1 overexpression can promote CRLR distribution on MG-63 cell membrane and enhance CGRP's promotion effect on MG-63 cell proliferation.
authors.affiliation: Department of Stomatology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Zhi-liang
authors.initials: ZL
authors.lastname: Zhao
authors.forename: Gang
authors.initials: G
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Ying-hui
authors.initials: YH
authors.lastname: Tan
mesh_terms: D001859:Bone Neoplasms / Q000378:metabolism / Q000473:pathology*; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005822:Genetic Vectors; D006801:Humans; D012516:Osteosarcoma / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D011897:Random Allocation; D058262:Receptor Activity-Modifying Protein 1 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: C545502:RAMP1 protein, human; D012333:RNA, Messenger; D058262:Receptor Activity-Modifying Protein 1; D015740:Calcitonin Gene-Related Peptide