The Effect of Cowanin on ERK1/2 and p-ERK1/2 Protein Expression in T47D Breast Cancer Cells by The Western Blot Method
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Abstract
Breast cancer in Indonesia occupies the first position in cancer cases that cause death in women. Recent studies have shown that cowanin is effective as a cytotoxic by inhibiting T47D cell cycle regulation in the G0-G1 phase. This study was conducted to determine the effect of cowanin on signal transduction by looking at the expression of extracellular regulated kinase (ERK)1/2 and phosphorylated-ERK1/2 (p-ERK1/2) proteins. The treatments were divided into 3 groups: negative control, cowanin, and doxorubicin. The administration of cowanin is IC50 11.11 g/mL, and doxorubicin is 0.125 g/mL. The western blot steps include lysing cells and separating them by gel electrophoresis, transferring protein bands from gel to membrane, and detecting protein bands by adding antibodies that react with the substrate. Parameters that will be observed are surface area and protein density using ImageJ. The results showed that cowanin could affect the expression of the ERK1/2 protein by significantly reducing the area (p<0.05) and the protein density of the ERK1/2 protein band significantly (p<0.05) followed by the post hoc Duncan test, there was a difference in the effect from treatment. Cowanin suppresses the expression of the ERK1/2 protein, which inhibits replication by decreasing the area and density of the ERK1/2 protein.
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