RESUMO
BACKGROUND: Picrasma quassioides (PQ) is a traditional Asian herbal medicine with anti-tumor properties that can inhibit the viability of HepG2 liver cancer cells. H-Ras is often mutated in liver cancer, however, the effect of PQ treatment on H-Ras mutated liver cancer is unclear. This study aimed to investigate the role of PQ on ROS accumulation and mitochondrial dysfunction in H-ras mutated HepG2 (HepG2G12V) cells. MATERIALS AND METHODS: PQ ethanol extract-induced HepG2G12V apoptosis was analyzed by the MTT assay, fluorescence microscopy, flow cytometry and western blotting. RESULTS: PQ treatment affected cell migration and colony formation in HepG2G12V cells. Cleaved-caspase-3, cleaved-caspase-9 and BCL2 associated agonist of cell death (BAD) expression levels were increased, while the levels of B-cell lymphoma-extra large (Bcl-xL) were decreased with PQ treatment. PQ treatment led to a reduction of H-Ras expression levels in liver cancer cells, thus reducing their abnormal proliferation. Furthermore, it led to increased expression levels of Peroxiredoxin VI, which regulates the redox signal in cells. CONCLUSION: Taken together these results provide a new functional significance for the role of PQ in treating HepG2G12V liver cancer.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Hepáticas/tratamento farmacológico , Mitocôndrias Hepáticas/efeitos dos fármacos , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas p21(ras)/genética , Espécies Reativas de Oxigênio/metabolismo , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Genes ras , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias Hepáticas/metabolismo , Picrasma/química , Proteínas Proto-Oncogênicas p21(ras)/biossínteseRESUMO
Contagious ecthyma (also known as orf) is an acute skin zoonosis caused by orf virus (ORFV), which affects sheep, goats and humans. As one of the typical species of the Parapoxvirus genus of the Poxviridae family, orf virus has distinctive and unique characteristics of these species. A range of immuno-modulatory/pathogenesis -related genes acquired by virus that function is to limit (at least transiently) the effectiveness of host immunity during its evolution. This review is aimed to describe the latest progress on the molecular characteristics of ORFV, and upon which we analyzed molecular mechanism of the immune escape designed and a set of strategies developed for ORFV to effective against immune clearance of the host. Known as an essential component in evolutionary system, host is regulated by ORFV for using in population evolution. By the ORFV evolutional immune regulation components and its effect approach, we can understand the viral biological characteristics of ORFV, and it is helpful for us to further study the counter-measures of this disease.
Assuntos
Ectima Contagioso/virologia , Evasão da Resposta Imune , Vírus do Orf/imunologia , Animais , Ectima Contagioso/imunologia , Regulação Viral da Expressão Gênica , Vírus do Orf/genética , Proteínas Virais/genética , Proteínas Virais/imunologiaRESUMO
AIM: The present study aims at preparation of CARP antibody and analysis of CARP expression pattern. METHODS: Through bioinformatics analysis the antigenicity of CARP, 279 bp cDNA fragment coding CARP N-terminal 93 amino acid was amplified by RT-PCR from mouse heart RNA, then cloned into pET-28b to construct the prokaryotic expression pET 28b-CARP N93. The plasmid was transformed into E.coli(BL21). The target fusion protein was expressed with induction of IPTG, and purified by affinity chromatography. The antiserum against CARP was obtained from the mice immunized with CARP. CARP expression patterns were examined by Western blot and immunofluorescence. RESULTS: Prepared CARP antibody; CARP specific expression in mouse heart and mainly localized in the nucleus in rat cardiac primary cells. CONCLUSION: Successfully generated CARP antibody and examined the expression pattern. The present study has laid a foundation for functional study of CARP.
