Impact of fragile histidine triad gene transfection on the proliferation and apoptosis of human colorectal cancer cell / 中华胃肠外科杂志
Chinese Journal of Gastrointestinal Surgery
; (12): 691-694, 2010.
Article
en Zh
| WPRIM
| ID: wpr-266286
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of fragile histidine triad (FHIT) gene transfection on human colorectal cancer cell line SW480 through up-regulation of caspase-8 expression.</p><p><b>METHODS</b>The eukaryotic expression plasmid containing FHIT, pRc/CMV2-FHIT was prepared and purified, and then identified by restrictive enzyme digestion. pRc/CMV2-FHIT was transfected into SW480 cells, and positive cell clones (SW480-FHIT, study group) were selected and amplified. Empty plasmid-transfected SW480 cells(SW480-pRc/CMV2, negative control) and normal SW480 cells (bland control) were used as control. Methyl thiazolyl tetrazolium (MTT) assay was used to test the changes in the proliferation of SW480 cells. Cell-cycle kinetics and apoptosis were analyzed by flow cytometry (FCM). The changes of pro-caspase-8, caspase-8 mRNA and caspase-8 relative activity were analyzed by Western blot, semi-quantitative RT-PCR and colorimetric assay with pan labeled substrate, respectively.</p><p><b>RESULTS</b>At 96 hours after transfection, cell inhibition rates of the study group and the negative control group were 71.7% and 16.9%. G0/G1 ratio was (63.2±3.5)% and (50.6±2.1)%, optical density of caspase-8 mRNA band 107 and 41, and relative activity of caspase-8 0.43 and 0.25, respectively. All the differences above were statistically significant (P<0.05). When FHIT inhibitor was added, the relative activity of caspase-8 decreased to 0.22, comparable to that in the control group.</p><p><b>CONCLUSIONS</b>FHIT gene transfection can significantly inhibit the proliferation and induce G0/G1 arrest in human colon cancer cell line SW480. The mechanism is related to the up-regulation of caspase-8 expression.</p>
Texto completo:
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Base de datos:
WPRIM
Asunto principal:
Patología
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ARN Mensajero
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Transfección
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Apoptosis
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Neoplasias del Colon
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Ácido Anhídrido Hidrolasas
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Línea Celular Tumoral
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Proliferación Celular
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Caspasa 8
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Genética
Límite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Gastrointestinal Surgery
Año:
2010
Tipo del documento:
Article