RESUMO
BACKGROUND: Triptolide is a structurally unique diterpene triepoxide with potent antitumor activity. However,the effect and mechanism of triptolide on hepatocellular carcinoma (HCC) is not well studied. METHODS: Cells were treated with triptolide, and the anti-HCC activity of triptolide was evaluated using flow cytometry, western blot, and xenograft studies. MicroRNA microarray and quantitative reverse-transcription polymerase chain reaction was used to identify differential microRNAs induced by triptolide. Chromatin immunoprecipitation assay was employed to study the interaction between c-Myc and genomic regions of miR106b-25. MicroRNAs overexpression and knockdown experiments were performed to determine the role of these microRNAs in triptolide-induced apoptosis. RESULTS: Triptolide inhibited cell proliferation and induced marked apoptosis in multiple HCC cell lines with different p53 status. Several signaling molecules involved in different pathways were altered after the treatment of triptolide. Xenograft tumor volume was significantly reduced in triptolide-treated group compared with vehicle control group. Two miRNA clusters, miR-17-92 and miR-106b-25, were significantly suppressed by triptolide, which resulted in the upregulation of their common target genes, including BIM, PTEN, and p21. In HCC samples, high levels of these miRNA clusters correlated with shorter recurrence free survival. Triptolide inhibited the expression of theses miRNAs in a c-Myc-dependent manner, which enhanced triptolide-induced cell death. We further showed that triptolide down-regulated the expression of c-Myc through targeting ERCC3, a newly identified triptolide-binding protein. CONCLUSIONS: The triptolide-induced modulation of c-Myc/miRNA clusters/target genes axis enhances its potent antitumor activity, which indicates that triptolide serves as an attractive chemotherapeutic agent against HCC.
Assuntos
Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Diterpenos/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , MicroRNAs/genética , Fenantrenos/farmacologia , Proteínas Proto-Oncogênicas c-myc/metabolismo , Animais , Antineoplásicos Alquilantes/farmacologia , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Morte Celular/efeitos dos fármacos , Morte Celular/genética , Linhagem Celular Tumoral , Proliferação de Células , DNA Helicases/genética , Proteínas de Ligação a DNA/genética , Modelos Animais de Doenças , Compostos de Epóxi/farmacologia , Perfilação da Expressão Gênica , Humanos , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Camundongos , Oncogenes , Interferência de RNA , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
OBJECTIVE: To observe in vitro and in vivo effects of triptolide on growth inhibition and apoptosis of osteosarcoma cells, and to further explore its correlated molecular mechanisms. METHODS: The growth inhibition effects of triptolide on osteosarcoma cells were detected using MTT assay. The apoptosis was detected using flow cytometry.The protein expressions of associated signals were detected using Western blot. The in vivo anti-osteosarcoma effects of triptolide were verified in osteosarcoma nude mice. The in vivo associated protein expressions were detected using immunohistochemistry (IHC). RESULTS: Triptolide could significantly inhibit the proliferation of various osteosarcoma cells. Besides, it could induce their apoptosis. Triptolide triggered the mitochondrial dependent apoptotic pathway, significantly inhibited the in vivo growth of osteosarcoma cells, and caused in vivo apoptosis. CONCLUSIONS: Triptolide induced apoptosis of osteosarcoma cells partially through activating mitochondria associated apoptosis signal pathway. Triptolide also induced apoptosis of osteosarcoma cells and inhibited their in vivo growth in nude mice.
