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    Evaluation of in vitro Anticancer Activity of Vulpinic Acid and its Apoptotic Potential Using Gene Expression and Protein Analysis
    (Assoc Pharmaceutical Teachers India, 2018) Kilic, Nil; Derici, Mehmet Kursat; Buyuk, Ilker; Aydin, Semra Soydam; Aras, Sumer; Cansaran-Duman, Demet
    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 mu M 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 real-time 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 Box 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.
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    Usnic acid causes apoptotic-like death in Leishmania major, L. infantum and L. tropica
    (Springer Heidelberg, 2018) Derici, Mehmet Kursat; Cansaran-Duman, Demet; Taylan-Ozkan, Aysegul
    Leishmaniasis, a deadly parasitic infection, threatens many people worldwide. Since the high cost, toxicity, and resistance are drawbacks of current treatment options, it is necessary to find safer and more effective new antileishmanial drugs. The aim of this study was to determine the antileishmanial activity of usnic acid and its apoptotic mechanism on Leishmania spp. promastigotes. The antileishmanial activity was evaluated by MTT assay and apoptosis-related gene expression was investigated by qRT-PCR. Usnic acid was to be effective against Leishmania major, L. infantum, and L. tropica promastigotes at IC50 =10.76 mu g/ml, 13.34 mu g/ml, and 21.06 mu g/ml, respectively. We also demonstrated a novel mechanism by which usnic acid inhibited proliferation and caused apoptosis; usnic acid upregulated p53, Bax, Casp-3, and Casp-9 gene expression and downregulated the level of Bcl-2 gene expression. Accordingly, the expression level of the P53 gene increased in L. major, L. infantum and L. tropica by 14.4-, 11.8-, and 9.5-fold, respectively, and in contrast, the Bcl-2 gene expression decreased in all three leishmaniasis by 0.8-, 0.8-, and 0.7-fold, respectively. The present study, therefore, revealed that usnic acid played a critical role in the usnic acid-induced apoptotic process in Leishmania species. Usnic acid is easily accessible and an inexpensive agent, and can be considered as an alternative therapeutic agent for Leishmania infections subject to further tests in animal models.