ABSTRACT
<p><b>OBJECTIVE</b>To investigate the anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc and its possible molecular mechanisms in vitro and in vivo.</p><p><b>METHODS</b>Transonic alcohol-chloroform extraction method was used to extract toosendanin from the bark of Melia toosendan Sieb. et Zucc, and the content of toosendanin in the crude extract was measured by high performance liquid chromatography (HPLC). Anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc were investigated in in vivo and in vitro studies. In the in vitro experiment, human hepatocellular carcinoma cell lines SMMC-7721 and Hep3B were co-incubated with toosendanin crude extract of different concentrations, respectively. In the in vivo experiment, BALB/c mice were subcutaneously inoculated with mouse hepatocellular carcinoma H22 cells and treated with crude extract.</p><p><b>RESULTS</b>HPLC revealed the content of toosendanin was about 15%. Crude extract from Melia toosendan Sieb. et Zucc inhibited cancer cells growth in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50, 72 h) was 0.6 mg/L for SMMC-7721 cells and 0.8 mg/L for Hep3B cells. Both high-dose [0.69 mg/(kg d)] and low-dose [0.138 mg/(kg d)] crude extract could markedly suppress cancer growth, and the inhibition rate was greater than 50%. Hematoxylin and eosin staining showed necrotic area in cancers and transmission electron microscopy displayed necrotic and apoptotic cancer cells with apoptotic bodies. Immunohistochemistry showed that the expression of Bax and Fas increased and the expression of Bcl-2 reduced.</p><p><b>CONCLUSIONS</b>Toosendanin extract has potent anti-cancer effects via suppressing proliferation and inducing apoptosis of cancer cells in vivo and in vitro. The mechanism of apoptosis involves in mitochondrial pathway and death receptor pathway.</p>
Subject(s)
Animals , Female , Male , Antineoplastic Agents , Pharmacology , Therapeutic Uses , Apoptosis , Carcinoma, Hepatocellular , Drug Therapy , Pathology , Cell Proliferation , Drugs, Chinese Herbal , Chemistry , Pharmacology , Therapeutic Uses , Immunohistochemistry , Liver Neoplasms , Drug Therapy , Pathology , Melia , Chemistry , Mice, Inbred BALB C , Mitochondria , Metabolism , Neoplasm Transplantation , Plant Extracts , Therapeutic Uses , Reference Standards , bcl-2-Associated X Protein , Metabolism , fas Receptor , MetabolismABSTRACT
Objective To investigate the effects and mechanism of Sufentanil on Hep3B cell viability in liver cancer.Methods Sufentanil of different concentrations (0.01,0.1,1,5,10,1 5 μmol/L)was used to treat Hep3B cells.The changes of cell cycle,apoptosis and protein expression were detected to explore the potential mechanisms by MTT method,flow cytometry and Western blot,respectively.Results Reduced viability of Hep3B cells,arrested cell cycle in the G0/G1 phase,decreased expression of survivin protein,and increased expression of caspase-3 protein were observed with the increase of Sufentanil concentration. Conclusion Sufentanil inhibited the viability of Hep3B cells through affecting cell cycle,promoting cell apoptosis and changing protein expressions of survivin and caspase-3.
ABSTRACT
PURPOSE: Hepatocellular carcinoma (HCC), a typical hypervasculized tumor is very sensitive to hypoxia and vascular endothelial growth factor (VEGF) has previously been identified to be up-regulated in response to hypoxia in several cell types. However, the molecular mechanisms by which hypoxia is sensed by the cells remain enigmatic. To investigate whether calcium and AP-1 are involved in hypoxia-sensing mechanism, we performed following experiments. MATERIALS AND METHODS: Hep3B cells were grown in hypoxic condition. To assess cell viability, MTT assay was performed. To investigate the effect of calcium and AP-1, northern blot analysis was performed after treatment with BAPTA/AM. RESULTS: The expression of VEGF was significantly up-regulated by hypoxia in Hep3B, hepatocellular carcinoma cell line. The increased expression of VEGF induced by hypoxia was blocked by the addition of BAPTA/AM, a cytosolic calcium chelator to the media. In addition, we found that the expression of c-jun protooncogene was also up-regulated by hypoxia. Hypoxic increase of c-jun expression was also normalized by the treatment with BAPTA/AM. CONCLUSION: These results suggest that the increased expression of VEGF by hypoxia is mediated through the calcium and c-jun signalling pathway in the Hep3B human hepatoma cell lines.