ABSTRACT
Background: Avimers are originally types of artificial proteins with multiple binding sites for specific binding to certain antigens. Various radioisotopes and nanoparticles link these molecules, which are widely used in early detection in tissue imaging, treatment and study on carcinogenesis. Among these, c-Met antagonist avimer [C426 avimer], with ability to bind the c-Met receptor of tyrosine kinase [RTK] is an attractive candidate for targeted cancer therapy. In this study, a novel traceable C426 avimer gene was designed and introduced by adding the 12nt tracer binding site encoded four specific amino acid residues at the C-terminal region of C426 avimer coding sequence
Methods: The 282 bp DNA sequence encoded 94aa avimer protein was synthesized and sub-cloned into prokaryotic pET26b expression vector. The expression of the mature peptide encoding the traceable avimer molecule was carried out in Escherichia coli strain BL21 using IPTG [Isopropyl beta-D-1-thiogalactopyranoside] induction process. The expression level of the 11 kDa traceable avimer was studied by SDS-PAGE, western blot and ELISA analysis
Results: Docking analysis of C426 avimer protein and its ligand c-Met showed that the traceability related changes happened at the best conformation and optimal energy. The SDS-PAGE, western blotting and ELISA analysis results demonstrated that the expression of the 11 kDa C426 avimer molecule was detectable without any degradation compared with the control group
Conclusion: Concerning the consequences of this work, this new approach can be widely used in the medical field and provide an opportunity to evaluate the affinity and traceability features
ABSTRACT
Aim: In this study we attempt to indicate anti-carcinogenic influence of ether extracted metabolites of Streptomyces Levis sp. on gene expression in colon cancer
Background: Colon cancer is one of the most prevalent cancers worldwide. In recent decades, researchers have been seeking the treatment for cancer. Natural products are valuable compounds with fewer side effects in comparison to chemotherapy drugs
Methods: Secondary metabolites were extracted with the inoculation of bacterial sample in Mueller Hinton Broth. MTT assay was done to evaluate the cytotoxicity effect of metabolites on SW480 cells. qRT-PCR was performed to observe effects of metabolites on Bcl-2, P53, SOX2, KLF4, Beta-Catenin, SMAD4, K-ras, BRAF genes expression in colon cancer
Results: The metabolites exhibited cytotoxic effects on colon cancer in a dose/time dependent manner [P < 0.001]. After 48 h treatment, fold expression of Bcl-2, SOX2, Beta-catenin, K-ras, BRAF genes fold of expression were decreased, whereas P53, KLF4, SMAD4 genes were increased in treated cells [P < 0.001]
Conclusion: These findings indicate that ether extracted metabolites of Streptomyces Levis ABRIINW111 have anti-carcinogenic effects on colon cancer
ABSTRACT
Because drug resistance is one of the most important problems in patients with chronic myeloid leukemia [CML], finding new anti-cancer drugs especially from natural sources is research priority. Therefore, in this study, anti-cancer effects of ethyl acetate soluble metabolite of Iranian native bacteria, Streptomyces calvus, were studied using human chronic myeloid leukemia K562 cells. In this experimental trail, ethyl acetate soluble metabolites were isolated from S. calvus bacteria. K562 cell line was treated by various concentrations of this metabolite for 12- 72 h intervals. Anti-proliferative effects of ethyl acetate soluble metabolite were studied by trypan blue exclusion test. Wright-Giemsa staining and latex particle phagocytosis assay were used to study differentiated cells. Ethyl acetate soluble metabolite induced growth inhibition in K562 cells in concentration and time- dependent manner. At the concentration of 200 ng/ml, the growth was inhibited 19-50% after 12-72h. Latex particle phagocytosis assay and Wright- Giemsa staining results revealed that K562 cells were differentiated toward monocytic- macrophagic lineage. According to growth inhibitory and differentiating effects of S.calvus metabolite in K562 cells, this metabolite can be proposed for more investigations in differentiation therapy of CML patients.