title: The anti-tumour activity of rLj-RGD4, an RGD toxin protein from Lampetra japonica, on human laryngeal squamous carcinoma Hep-2 cells in nude mice.
abstract: OBJECTIVE
  The objective of this study is to investigate the antiproliferative activity and mechanism of integrin-binding rLj-RGD4 in a Hep-2 human laryngeal carcinoma-bearing nude mouse model.
  
  
  METHODS
  Human laryngeal squamous carcinoma cells (Hep-2) were inoculated subcutaneously into the axilla of nude mice to generate a Hep-2 human laryngeal carcinoma-bearing nude mouse model. When the Hep-2 xenograft model was successfully established, the animals were randomly separated into five groups. Three groups were treated with different dosages of rLj-RGD4. Cisplatin was administered to the positive control group, and normal saline (NaCl) was administered to the negative control group for 3 weeks. The body weights and the survival of the nude mice were evaluated, and the volumes and weights of the solid tumours were measured. The mechanism underlying rLj-RGD4 inhibition of tumour growth in transplanted Hep-2 human laryngeal carcinoma-bearing nude mice was evaluated by haematoxylin-eosin (HE) staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL), measurement of intratumoural microvessel density (MVD), Western blotting, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR).
  
  
  RESULTS
  The tumour volumes and weights of the treatment groups were reduced compared with the model group, and survival times were improved by rLj-RGD4 treatment in Hep-2 human laryngeal carcinoma-bearing nude mice. The number of apoptotic Hep-2 human cells and intratumoural MVD significantly decreased after the administration of rLj-RGD4. In the xenograft tissue of animals treated with rLj-RGD4, FAK, PI3K, and Akt expression was unaltered, whereas P-FAK, P-PI3K, Bcl-2, P-Akt, and VEGF levels were down-regulated. In addition, activated caspase-3, activated caspase-9, and Bax levels were up-regulated.
  
  
  CONCLUSIONS
  rLj-RGD4 exhibits potent in vivo activity and inhibits the growth of transplanted Hep-2 human laryngeal carcinoma cells in a nude mouse model. Thus, these results indicate that the recombinant RGD toxin protein rLj-RGD4 may serve as a potent clinical therapy for human laryngeal squamous carcinoma.
authors.affiliation: Department of Pharmacology, Dalian Medical University, Dalian, Liaoning Province, 116044, China.
authors.forename: Fangyu
authors.initials: F
authors.lastname: Shao
authors.affiliation: Department of Otorhinolaryngology and Head and Neck Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning, 116011, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Lv
authors.affiliation: School of Life Sciences, Liaoning Normal University, Dalian, Liaoning Province, 116029, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Department of Pharmacology, Dalian Medical University, Dalian, Liaoning Province, 116044, China.
authors.forename: Junshu
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Department of Pharmacology, Dalian Medical University, Dalian, Liaoning Province, 116044, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, Dalian Medical University, Dalian, Liaoning Province, 116044, China. Electronic address: lv_li@126.com.
authors.forename: Li
authors.initials: L
authors.lastname: Lv
authors.affiliation: School of Life Sciences, Liaoning Normal University, Dalian, Liaoning Province, 116029, China. Electronic address: jihongwang999@hotmail.com.
authors.forename: Jihong
authors.initials: J
authors.lastname: Wang
mesh_terms: D020816:Amino Acid Motifs; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry / Q000627:therapeutic use*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001835:Body Weight / Q000187:drug effects; D002294:Carcinoma, Squamous Cell / Q000098:blood supply / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D029941:Fish Proteins / Q000008:administration & dosage / Q000009:adverse effects / Q000235:genetics / Q000627:therapeutic use*; D005397:Fish Venoms / Q000737:chemistry*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006258:Head and Neck Neoplasms / Q000098:blood supply / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007798:Lampreys; D007822:Laryngeal Neoplasms / Q000098:blood supply / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008387:Marine Toxins / Q000008:administration & dosage / Q000009:adverse effects / Q000235:genetics / Q000627:therapeutic use*; D008819:Mice, Nude; D055806:Microvessels / Q000187:drug effects / Q000473:pathology; D009842:Oligopeptides / Q000737:chemistry; D011897:Random Allocation; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry / Q000627:therapeutic use; D000077195:Squamous Cell Carcinoma of Head and Neck; D016019:Survival Analysis; D047368:Tumor Burden / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
chemical_list: D020533:Angiogenesis Inhibitors; D029941:Fish Proteins; D005397:Fish Venoms; D008387:Marine Toxins; D009842:Oligopeptides; C000611447:RGD4 protein, Lampetra japonica; D011994:Recombinant Proteins; C047981:arginyl-glycyl-aspartic acid
keywords: Anti-tumour; Hep-2; Laryngeal squamous carcinoma; Nude mice; rLj-RGD4