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1.
Biomedical and Environmental Sciences ; (12): 391-399, 2011.
Artigo em Inglês | WPRIM | ID: wpr-306847

RESUMO

<p><b>OBJECTIVE</b>In order to investigate the potential mechanisms in troglitazone-induced apoptosis in HT29 cells, the effects of PPARγ and POX-induced ROS were explored.</p><p><b>METHODS</b>[3- (4, 5)-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay, Annexin V and PI staining using FACS, plasmid transfection, ROS formation detected by DCFH staining, RNA interference, RT-PCR & RT-QPCR, and Western blotting analyses were employed to investigate the apoptotic effect of troglitazone and the potential role of PPARγ pathway and POX-induced ROS formation in HT29 cells.</p><p><b>RESULTS</b>Troglitazone was found to inhibit the growth of HT29 cells by induction of apoptosis. During this process, mitochondria related pathways including ROS formation, POX expression and cytochrome c release increased, which were inhibited by pretreatment with GW9662, a specific antagonist of PPARγ. These results illustrated that POX upregulation and ROS formation in apoptosis induced by troglitazone was modulated in PPARγ-dependent pattern. Furthermore, the inhibition of ROS and apoptosis after POX siRNA used in troglitazone-treated HT29 cells indicated that POX be essential in the ROS formation and PPARγ-dependent apoptosis induced by troglitazone.</p><p><b>CONCLUSION</b>The findings from this study showed that troglitazone-induced apoptosis was mediated by POX-induced ROS formation, at least partly, via PPARγ activation.</p>


Assuntos
Humanos , Antineoplásicos , Farmacologia , Apoptose , Cromanos , Farmacologia , Citocromos c , Genética , Metabolismo , Regulação Neoplásica da Expressão Gênica , Células HT29 , PPAR gama , Metabolismo , Prolina Oxidase , Metabolismo , Espécies Reativas de Oxigênio , Metabolismo , Tiazolidinedionas , Farmacologia
2.
Medical Journal of Chinese People's Liberation Army ; (12)2001.
Artigo em Chinês | WPRIM | ID: wpr-552990

RESUMO

To explore the the effect of RSV M 2-1 gene expression on the growth of human lung adenocarcinoma PAa cell line, the recombinant RSV M 2-1 gene eukaryotic plasmid PXJ 41/ M 2-1 was transfected to the human lung adenocarcinoma PAa cell line. Expression of the M 2-1 gene was examined by RT PCR and Western blot. The growth of the human lung adenocarcinoma PAa cell was observed by MTT curve, flow cytometry, the capacity of inherence, colony forming units and inoculation of nude mouse. The results showed that ①The desired fragments of M 2-1 gene were digested by restriction enzyme and RT PCR, respectively. ②The bands of M 2-1 gene protein were found by Western blot. ③ The ratio of transfected PAa cells in S phase of cell cycle was decreased, but increased in G 2 /M phase. Colony forming efficiency and units of the transfected PAa cell were increased, but the capacity of its inherence was decreased. ④Tumorogensising time of the transfected PAa cell in nude mouse was delayed, but its growing speed was increased. The tumor tissue transformed to some extent from adenocarcinoma to squamous carcinoma was found in the nude mouse. It is suggested that the M 2-1 gene may promote the growth of PAa cells, which may implicate some relationship between RSV and lung cancer.

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