RESUMEN
Crude extract of Scutellaria baicalensis (S. baicalensis) has cytotoxic effect on human myelogenous leukemia cells (HL-60). We invesigated which compound from the crude extract is responsible for the cytotoxic effect on HL-60 cells. We identified 29 compounds from the crude extract using high performance liquid chromatography mass spectrometry (HPLC/MS). Two of the compounds, baicalin and wogonoside, are converted to baicalein and wogonin, respectively, after treatment with beta-glucuronidase. We observed a dose-dependent reduction in cell viability when cells with either wogonin or aqueous extract of S. baicalensis. Several of the apoptotic features including deoxyribonucleic acid (DNA) fragmentation and increased caspase-3 activity were found in cells treated with wogonin and aqueous extract. The changes were associated with down-regulation of Bcl-2, and not Bax. Furthermore, treatment of HL-60 cells with wogonin or S. baicalensis led to the inhibition of human telomerase reverse transcriptase (hTERT), human telomerase-associated protein 1 (hTP1) and c-myc messenger ribonucleic acid (m-RNA) expression. Wogonin and S. baicaleisis down-regulated the telomerase activity. Our findings suggest that wogonin may be the major compound in S. baicalensis responsible for HL-60 growth inhibition in vitro. The inhibition of HL-60 cell growth is mediated partly through the induction of Bax/Bcl-2 apoptosis and by telomerase inhibition through suppression of c-myc, which is a promoter of hTERT.
Asunto(s)
Antineoplásicos Fitogénicos/uso terapéutico , Apoptosis/efectos de los fármacos , Flavanonas/uso terapéutico , Leucemia Promielocítica Aguda/tratamiento farmacológico , Extractos Vegetales/uso terapéutico , Telomerasa/antagonistas & inhibidores , Antineoplásicos Fitogénicos/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo , Flavanonas/farmacología , Células HL-60 , Humanos , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Extractos Vegetales/química , Extractos Vegetales/farmacología , ARN Mensajero/metabolismo , Scutellaria baicalensis/química , Telomerasa/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteína X Asociada a bcl-2/antagonistas & inhibidoresRESUMEN
BACKGROUND: This study was designed to obtain the chemical fingerprint and to investigate the effect of Phyllanthus urinaria on telomerase activity and apoptotic pathways in the human nasopharyngeal carcinoma cell line (NPC-BM1). MATERIALS AND METHODS: The polyphenol compounds in P. urinaria were investigated by HPLC/MS. Cell viability with the treatment of P. urinaria, gallic acid, ellagic acid, quercetin and cisplatin was detected by MTT assay. TUNEL assay, DNA fragmentation analysis and caspase3 activity were used to confirm apoptotic changes. Telomerase activity was determined using the TRAP assay. RNA isolation and RT-PCR were used to analyze the related genes expression. All experiments on treatments with P. urinaria from 0-3 mg/ml were carried out for 24 h. RESULTS: 5 major compounds including gallic acid, brevifolin carboxylic acid, corilagin, phyllanthusiin C and ellagic acid were identified as a plant fingerprint by HPLC/MS. With the MTT assay, we demonstrated that P. urinaria, gallic acid and ellagic acid reduce cell viability. The apoptosis features showed DNA fragmentation and increased caspase-3 activity associated with the down-regulation of Bcl-2, but not of Bax, p53, and PCNA (proliferating cell nuclear antigen) in P. urinaria-treated NPC-BM1 cells. Furthermore, treatment of NPC-BM1 cells led to an inhibition of hTERT (human telomerase reverse transcriptase), hTP1 (human telomerase-associated protein 1) and c-myc mRNA expression and to decreased telomerase activity. CONCLUSION: This study suggests that P. urinaria induces the death of NPC-BM1 cells in vitro through the induction of apoptosis and inhibited telomerase activity.
Asunto(s)
Apoptosis/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Phyllanthus/química , Extractos Vegetales/farmacología , Antineoplásicos/farmacología , Antioxidantes/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Cisplatino/farmacología , Medicamentos Herbarios Chinos/química , Ácido Elágico/farmacología , Ácido Gálico/farmacología , Genes bcl-2/genética , Genes myc/genética , Humanos , Espectrometría de Masas , Neoplasias Nasofaríngeas , Extractos Vegetales/química , Quercetina/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , TelomerasaRESUMEN
Seven new cytotoxic xenicane-type diterpenoids, 9-deoxyxeniloide-E (1), 9-deoxy-7,8-epoxyxeniloide-E (2), xeniolide-G (3), 9-deoxyxenialactol-C (4), xenibecin (5), xeniolide-H (6), and xenitacin (7), were isolated from the methylene chloride solubles of the Formosan soft coral Xenia umbellata. The structures were elucidated by 1D and 2D NMR spectral analysis, and their cytotoxicity against selected cancer cells was measured in vitro.
Asunto(s)
Antozoos/química , Antineoplásicos/aislamiento & purificación , Diterpenos/aislamiento & purificación , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Cromatografía en Capa Delgada , Neoplasias del Colon , Diterpenos/química , Diterpenos/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Leucemia P388 , Neoplasias Pulmonares , Ratones , Conformación Molecular , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Estereoisomerismo , Taiwán , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Three new cytotoxic prostanoids, claviridenone E-G (1-3), and three new cytotoxic steroids, stoloniferone E-G (4-6), were isolated from the methylene chloride solubles of the Formosan soft coral Clavulariaviridis. A cytotoxic cembranoid, claviolide (7), was isolated from the methylene chloride solubles of the Formosan soft coral Clavularia violacea. The structures were elucidated by 1D and 2D NMR spectral analysis, and their cytotoxicity against selected cancer cells was measured in vitro.