Effect of siRNA targeting centromere protein-A gene on biological behavior of HepG2 cells / 中华病理学杂志
Chinese Journal of Pathology
; (12): 124-128, 2008.
Article
em Zh
| WPRIM
| ID: wpr-349960
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To study the influence of siRNA inhibition of CENP-A expression on the biological behavior of HepG2 cells.</p><p><b>METHODS</b>Three pairs of 21 bp reverse repeated motifs of CENP-A target sequence with 9 spacer were synthesized and inserted into vector pSilencer 2.1-U6 neo to generate siRNA eukaryotic expression plasmids. After stable transfection into HepG2 cells, cell growth, apoptosis, cell cycles and plate clone forming efficiency were investigated. Expressions of CENP-A mRNA was monitored by the reverse transcriptase polymerase chain reaction (RT-PCR). The protein expression of CENP-A, bcl-2, Bax, p53, p21waf1 and mdm2 were detected by Western-blotting.</p><p><b>RESULTS</b>Two eukaryotic expression plasmids with significant siRNA specific inhibition to the CENP-A gene were created. Compared with control cells, HepG2 cells transfected with the constructs showed G1 phase delay (P < 0.01) and cell number decrease in the S phase (P < 0.001), along with an increased apoptotic rate (P = 0.003), significant increase of Bax expression and decreased bcl-2 expression (P< or =0.001). The protein expressions of p21waf1 was higher and mdm2 was lower than those of the control groups. However, the wild type p53 protein expression was not effected by CENP-A siRNA.</p><p><b>CONCLUSIONS</b>An altered expression of CENP-A may be related to the proliferation of hepatocellular carcinoma through cell cycle regulation involving an altered bcl-2/Bax expression, that may be p53 independent.</p>
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Base de dados:
WPRIM
Assunto principal:
Patologia
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Farmacologia
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Autoantígenos
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Células Tumorais Cultivadas
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Proteínas Cromossômicas não Histona
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Regulação Neoplásica da Expressão Gênica
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Carcinoma Hepatocelular
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Reação em Cadeia da Polimerase Via Transcriptase Reversa
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RNA Interferente Pequeno
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Interferência de RNA
Limite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Pathology
Ano de publicação:
2008
Tipo de documento:
Article