Modulation of C5a-C5aR interactions against murine mammary cancer cell line

Abstract

Background: Complement C5a is a potent in ammatory chemoattractant and might be a bene cial therapeutic target for the induction of an effective anti-tumour response. C5a agonist and antagonist modulation have demonstrated to have either promotes or inhibits tumour development, EMT6 murine mammary cancer cells through both in vitro and in vivo studies.Methods: For the in-vitro studies, Alamar Blue cell viability assay was used for cell viability determination and immuno uorescence assay was used to determine the location of C5aR expression on EMT6 cell line. For the in-vivo experiment, female Balb/c mice were subcutaneously injected with EMT6 tumour cells and subsequently treated with both C5aR agonist and antagonist peptides. At the end of in-vivo study period of 14 days, liver and tumour samples were obtained for an ELISA assay to quantify the levels of TNF-α, caspase-3, C5a and VEGF-A signals following the treatment with both C5aR agonist and antagonist. One-way ANOVA test was performed to evaluate the differences between the mean of treatments given in a group and that of the negative control Results: The in-vitro experiment revealed an expression of C5aR was found on the cell membrane of the EMT6 cells and treatment with EP54; which is the C5aR agonist has shown low cell viability after 48 hours post-treatment. For the in-vivo experiment, the ELISA assay outcome have shown that EP54 signi cantly promote high numbers of circling of signaling proteins except for VEGF-A, suggesting that the C5aR agonist modulation might inhibits tumour development and also trigger the induction of apoptosis. Conclusion:C5aR modulation through the in uence of EP54 agonism may have bene cial effects in terms of reduction of the tumour size and attenuation of its development.