[HBx gene down-regulates miR-192 expression and inhibits apoptosis of human hepatoma cell line HepG2].
Zhonghua Gan Zang Bing Za Zhi
; 19(11): 857-60, 2011 Nov.
Article
em Zh
| MEDLINE
| ID: mdl-22433310
ABSTRACT
OBJECTIVE:
To explore the mechanism by which HBV X gene(HBx) inhibits apoptosis of human hepatoma cell line HepG2 in terms of miRNA.METHODS:
Three cell lines were prepared HepG2 cells stably transfected with HBx (HepG2/HBx), HepG2 cells stably transfected with pcDNA3.1 (HepG2/pcDNA3.1) and HepG2 cells. Flow cytometry was adopted to measure the apoptosis of these three cells and Taqman fluorescence quantitative PCR was used to examine miR-192 expression. After HepG2 cells was transfected with miR-192, the apoptosis was analyzed by flow cytometry and the expressions of p53 and PUMA at mRNA and protein levels were evaluated by SYBR Green quantitative PCR and Western blot, respectively.RESULTS:
Compared with HepG2/pcDNA3.1 cells (11.46% ± 0.69%) and HepG2 cells (12.5% ± 0.66%), the apoptosis rate of HepG2/HBx cells (2.37% ± 0.35%) was significantly reduced (F = 171.722, P < 0.01). The level of miR-192 was 49.1% ± 5.9% in HepG2 cells, which was dramatically down-regulated (F = 14.319, P = 0.019) as compared to the other two groups (HepG2/pcDNA3.1 98.0% ± 8.9%; HepG2 100%). Compared with HepG2 cells transfected with miR-NC (10.74% ± 1.15%), transfection of miR-192 into HepG2 cells led to increased apoptosis (15.74% ± 1.17%) (F = 18.415, P = 0.013) and higher p53 and PUMA expressions at mRNA (p53 1.68 ± 0.12 vs 0.90 ± 0.09, F = 43.115, P = 0.003, PUMA 1.66 ± 0.10 vs 0.98 ± 0.06, F = 22.541, P = 0.009) and protein (p53 3.07 vs 1, PUMA 2.13 vs 1) levels.CONCLUSION:
HBx could inhibit apoptosis of HepG2 cells through down-regulation of miR-192 which induces apoptosis of HepG2 cells.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Transativadores
/
Vírus da Hepatite B
/
Apoptose
/
Carcinoma Hepatocelular
/
MicroRNAs
/
Neoplasias Hepáticas
Limite:
Humans
Idioma:
Zh
Revista:
Zhonghua Gan Zang Bing Za Zhi
Assunto da revista:
GASTROENTEROLOGIA
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
China