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A Novel Anti-Tumor Protein From Calloselasma Rhodostoma Venom In Vietnam

Kiem X. Trinh, Thuc v Pham, Long X Trinh, My T Chu, Thuc M Vu, Khanh B Nguyen, Francis Markland. Jr, Stephen Swenson, Radu X Radu Minea, et al

Abstract

Abstract

Introduction: Vietnam is a tropical and agricultural country. Annually, there are 30,000 cases of snake bite. Two venomous snake families cause the big medical problem. In this, Calloselasma rhodostoma (CR) is the most dangerous snake. Therefore, since 2001, the scientific research collaborations between Vietnam (VN) and Uiversity of Southern California (USC) were established. Since 2014 this project has been approved by VN government. The aims of the 1st project are to establish the technological process for purification of disintegrin from CR venom of VN (CRd.VN), to determin the molecular weigh, structure and its biological antitumor activities. Methods: The process of collection, lyophilization of CR venom from VN. Protein concentration of CR venom was determined by BCA assay. High Performance Liquid Chromatography (HPLC), SDS-PAGE, Mass spectrometry (MS)analysis and sequencing by tryptic digestion were used for purification of CRd.VN and its molecular weight (MW) and structure. Standard cell biology methods were employed to characterize CRd’s abilities (in vitro) to inhibit platelet aggregation, adhesion, migration and invasion of tumor cells. Its anti-cancer activity in a breast cancer (BC) murine model (in vivo ) was tested.The resuls: Peak No:7 of HPLC (CRd.VN), showed a single ≈10 kDa band on SDS-PAGE gel. CRd.VN’s MW, structure and the sequence are 7.33 kDa, a monomer containing 68 amino acids with an RGD motif (position 49-51) and 6 disulfide bonds. The anti-cancer activities of CRd.VN are very strong. Conclusion: We have shown that CRd.VN is a possible anti-tumor agent with clinical potential. However, further research is required on CRd.VN recombinant production, preliminary pharmacokinetics/ toxicology properties and anti-tumor activities.
Keyword: CRd anti-cancer.

(1):Vietnam Institute on Toxicology (VST).
(2):University of Medicine & Pharmacy of Haiphong.
(3):Biochemistry and Molecular Biology, Keck School of Medicine of USC.

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