title: Expression of complement C5a receptor and the viability of 4T1 tumor cells following agonist-antagonist treatment.
abstract: BACKGROUND
  Complement system is theoretically believed to halt the progression of tumor by the activity of C5a/CD88. Protein C5a is a potent pro.inflammatory mediator that activates the complement system by binding to its receptor.
  
  
  OBJECTIVE
  The purpose of this study is to determine the expression of the anaphylatoxin C5a receptor on 4T1 cell line and to study the viability of the cells after being treated with the C5a peptides.
  
  
  METHODS
  The cells 4T1 had undergone immunofluorescence staining, conventional polymerase chain reaction (PCR) and real-time PCR for the expression of determination part. Whereas Alamar Blue and MTT assays were conducted for the viability study of the cells.
  
  
  RESULTS
  The cells showed positive result in expressing the receptor of the C5a through immunostaining and PCR. The CT value recorded at initial dilution was 22.24. In cell viability assay, the cell was treated with C5a peptides, PMX205 and EP54. The purpose of this treatment was to see whether C5a had a direct effect on the cell itself using both assays. The result showed that PMX205, which is an antagonist, gave more effects towards the cell as compared with the treatment of EP54.
  
  
  CONCLUSIONS
  This experiment shows the presence of C5a receptor on 4T1 cell line. We believe that the antagonist peptide is eligible to be used widely in cancer immunotherapy field; but in vivo studies need to be carried out first in the future, as it will determine how these drugs affect the tumor cell growth.
authors.affiliation: Department of Veterinary Pre-Clinical Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
authors.forename: Nurneqman Nashreq
authors.initials: NN
authors.lastname: Kosni
authors.affiliation: Department of Veterinary Pre-Clinical Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
authors.forename: Norhaifa
authors.initials: N
authors.lastname: Ganti
authors.affiliation: Department of Veterinary Pre-Clinical Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
authors.forename: Mohd Hezmee Mohd
authors.initials: MH
authors.lastname: Noor
authors.affiliation: Department of Veterinary Pre-Clinical Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
authors.forename: Intan Shameha Abdul
authors.initials: IS
authors.lastname: Razak
authors.affiliation: Department of Veterinary Pre-Clinical Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
authors.forename: Mohd Mokrish Md
authors.initials: MM
authors.lastname: Ajat
authors.affiliation: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine; Institute of BioScience, Universiti Putra Malaysia, Selangor, Malaysia.
authors.forename: Abdul Rahman
authors.initials: AR
authors.lastname: Omar
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival; D015936:Complement C5a / Q000627:therapeutic use*; D005260:Female; D005455:Fluorescent Antibody Technique; D008325:Mammary Neoplasms, Experimental / Q000188:drug therapy* / Q000276:immunology / Q000473:pathology; D051379:Mice; D010446:Peptide Fragments / Q000627:therapeutic use*; D010456:Peptides, Cyclic / Q000627:therapeutic use*; D016133:Polymerase Chain Reaction; D044087:Receptor, Anaphylatoxin C5a / Q000819:agonists / Q000032:analysis* / Q000037:antagonists & inhibitors
chemical_list: D010446:Peptide Fragments; D010456:Peptides, Cyclic; D044087:Receptor, Anaphylatoxin C5a; C105322:complement C5a, (65-74), Tyr(65)-Phe(67)-Pro(69,71)-Ala(73)-; C504156:hydrocinnamate-cyclo(ornithyl-prolyl-cyclohexylalanyl-tryptophyl-arginyl); D015936:Complement C5a