Growth Inhibition and Apoptosis Induction by -Cyanidan-3-ol in Hepatocellular CarcinomaReport as inadecuate

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The objective of this study was to evaluate the cytotoxicity of +-cyanidan-3-ol CD-3 in human hepatocellular carcinoma cell line HepG2 and chemopreventive potential against hepatocellular carcinoma HCC in Balb-c mice. The HepG2 cell line was treated with CD-3 at various concentrations and the proliferation of the HepG2 cells was measure by 3-4,5-dimethylthiazol-2-yl-2,5-diphenyl-tetrazolium bromide MTT, sulforhodamine B SRB and lactate dehydrogenase LDH assays. Cell apoptosis was detected by Hoechst 33258 HO, Acridine orange-ethylene dibromide AO-EB staining, DNA fragmentation analysis and the apoptosis rate was detected by flow cytometry. The HCC tumor model was established in mice by injecting N-nitrosodiethylamine-carbon tetrachloride NDEA-CCl4 and the effect of CD-3 on tumor growth in-vivo was studied. The levels of liver injury markers, tumor markers, and oxidative stress were measured. The expression levels of apoptosis-related genes in in-vitro and in vivo models were determined by RT-PCR and ELISA. The CD-3 induced cell death was considered to be apoptotic by observing the typical apoptotic morphological changes under fluorescent microscopy and DNA fragmentation analysis. Annexin V-PI assay demonstrated that apoptosis increased with increase in the concentration of CD-3. The expression levels of apoptosis-related genes that belong to bcl-2 and caspase family were increased and AP-1 and NF-κB activities were significantly suppressed by CD-3. Immunohistochemistry data revealed less localization of p53, p65 and c-jun in CD-3 treated tumors as compared to localization in NDEA-CCl4 treated tumors. Taken together, our data demonstrated that CD-3 could significantly inhibit the proliferation of HepG2 cells in-vitro and suppress HCC tumor growth in-vivo by apoptosis induction.

Author: Jitender Monga, Saurabh Pandit, Rajinder Singh Chauhan, Chetan Singh Chauhan, Shailender Singh Chauhan, Manu Sharma



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