DEVELOPING A SYSTEM FOR ANALYZING THE DOWNSTREAM EFFECTS OF CCL5 DERIVATIVES ON THE CCR5 PATHWAY

Abstract

Background: Various types of diseases are known to be influenced by the ССL5/ССR5 pathway, including cancers and HIV. Inhibiting the attachment of CCL5 to CCR5 has shown promise in preclinical studies as a way to suppress the growth of the tumor and improve outcomes in cancer patients. In this study, we want to observe how p21, p53, and Mdm2 protein expressions that play important roles in cell growth will change with different modifications of CCL5. We aim to check the level of expression of these proteins in the cell line derived from brеаst саnсеr - МСF-7.

Маtеrіаls & Меtһоds: МСF-7, МDА-МВ-231, СНО/CCR5, and СНО/WT cell lines were cultured for further RNA isolation, RT-PCR, and qPCR experiments. RNA isolation and RT-PCR were performed in order to check whether the expected genes exist. In order to check the expression levels of three target proteins quantitative PCR was done.

Results: RT-PCR interpretation on a gel showed that cell lines express p21, Mdm2, and GAPDH genes, but failed to show p53. Only MDA-MB-231 cells were positive for CCR5 on a gel. qPCR analysis failed to determine Ct values for GAPDH and p53. That is why we were unable to calculate gene expression values for target genes and the experiment should be repeated.