Detection of anticancer and apoptotic effect of the produced IgYs against the three extracellular domain of human DR5 protein

ABSTRACT

TNF alpha cytokine family in the body plays divers' roles in the cellular events such as cell proliferation, differentiation, necrosis, septic shock and apoptosis. In response to TNF therapy, several cell signaling pathways activated in cells which in different manners can lead to apoptosis or necrosis. However induction of apoptosis is depended on one of its important members, TRAIL and its receptors that will be followed by apoptosis activity. Tumor necrosis factor-related apoptosis-inducing ligand [TRAIL] and especially the DR5, is generating considerable interests as a possible anticancer therapeutic agent because of its selective activation in apoptosis of this receptor as a superior affinity to ligands. The study was performed in invitro assay and the anticancer effects of the produced antibodies were assumed by MTT and flowcytometric methods. In the first step for immunization of the hens, three selective small peptides from extracellular domain of DR5 which were chemically synthesized, injected to hens and after the proper immunization of them, IgYs were extracted from the egg yolk. After assumption of specificity of the purified IgYs against the whole DR5 protein, they were performed in MTT assay and flowcytometric colorimeter. After confirmation of synthesized peptides they were injected to hens with Fround`s complete adjuvant. With completing the immunization procedure the specificity of purified IgYs were confirmed by ELISA. The antibodies were significantly killed the MCF7 breast cancer cells, but had divers affect [proliferative] on normal hepatocyte cells. Additionally, significantly they induced apoptosis on the cancerous cells in contrast to control cells. The results clearly demonstrated that the produced IgYs with reduced cost and time managing could remarkably use as an effective anticancer drug