Анатомия и физиология: текущие исследования
Открытый доступ
ISSN: 2161-0940
ISSN: 2161-0940
Kiem X Trinh, Pham Van Thuc, Chu Thi My, Long X Trinh, Thuc Thi Minh Vu, Bao Khanh Nguyen, Francis Markland J, Stephen Sweenson and Radu Minea
Aim: 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 University 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 dsintegrin from CR venom of VN (CRd.VN), to determine the molecular weight, structure and its biological antitumor activities.
Methods: The process of collection, lyophilisation 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.
Results: Peak 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 anticancer 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 recombinant production, preliminary pharmacokinetics/ toxicology properties and antitumor activities.