Evaluation of in vitro Anticancer Activity of Vulpinic Acid and its Apoptotic Potential Using Gene Expression and Protein Analysis

Abstract:

Lichens and their secondary metabolite are still among the many unexamined natural sources in the drug industry. This study was designed to evaluate the cytotoxic effects of vulpinic acid lichen secondary metabolite and 6.25, 12.5, 25, 50, 100, 200 and 400 μM concentrations that treat cancer cell lines (CaCo2, HepG2, Hep2C, RD Wehi) and normal cells (Vero and L929) by MTT assay. The aim of this study was to determine the apoptotic effect of vulpinic acid on a molecular level. The determination of apoptotic molecular patterns of vulpinic acid was performed on western blot and quantitative realtime PCR (qRT-PCR) analysis. In our study, transcriptome changes on both gene and protein levels showed similar results. The determination of, mRNA levels of Bax, Bcl-2 and P53 genes were showed by qRT-PCR in cancer and normal cell lines. The results of the study showed that IC₅₀ value of vulpinic acid altered the mRNA levels of Bax, Bcl- 2 and P53 genes in all examined cancer cells when compared to the untreated cells. When the mRNA levels of the examined genes were compared, it was observed that Bax gene showed more expression increase in all cell lines when compared to Bcl-2 and P53 genes. This is the first evaluation of the apoptotic effect of vulpinic acid secondary metabolite on mRNA levels. The current study highlights some points regarding vulpinic acid and its use for cancer treatment.

Key words: Vulpinic acid, Apoptotic effect, qRT-PCR, Western blot.