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
in Chinese
| WPRIM
| ID: wpr-349960
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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Pharmacology
/
Autoantigens
/
Tumor Cells, Cultured
/
Chromosomal Proteins, Non-Histone
/
Gene Expression Regulation, Neoplastic
/
Carcinoma, Hepatocellular
/
Reverse Transcriptase Polymerase Chain Reaction
/
RNA, Small Interfering
/
RNA Interference
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Pathology
Year:
2008
Type:
Article
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