Overexpression of bcl-2 protects hepatoma cell line HCC-9204 from ethanol-induced apoptosis / 中华医学杂志(英文版)
Chinese Medical Journal
;
(24): 8-11, 2002.
Article
in English
| WPRIM
| ID: wpr-308134
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of overexpression of bcl-2 on ethanol-induced apoptosis of primary hepatocellular carcinoma (HCC) cells.</p><p><b>METHODS</b>The retrovirus expression vector pDOR-SB containing human bcl-2 cDNA was introduced into a human HCC cell line HCC-9204 by liposome-mediated transfection. pDOR-transfected and non-transfected HCC-9204 cells were used as controls. The expression of Bcl-2 protein by transfected HCC-9204 cells was detected by the immunohistochemical method. Then the cells were cloned with the limited dilution method continually until a monoclonal cell strain whose positive rate of Bcl-2 protein was 100% detected by flow cytometry was obtained. The killing rates of cells were detected by Methabenzthiazuron assay after the treatment of 6% ethanol for 6 h. The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry.</p><p><b>RESULTS</b>Most of the pDOR-SB-transfected cells demonstrated Bcl-2 positive signals, while no signal was found in the controls. The positive rate of Bcl-2 protein detected by flow cytometry in the obtained monoclonal cell strain, which was named HCC-bcl2, was 100% after the cells had been cloned 3 times continually. The killing rate, TUNEL index and the scale of sub-G1 apoptotic peak in HCC-bcl2 cells were all significantly lower than those in the control cells.</p><p><b>CONCLUSION</b>Overexpression of Bcl-2 protein suppresses ethanol-induced apoptosis of the HCC cell line HCC-9204.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Pharmacology
/
Physiology
/
Tumor Cells, Cultured
/
Transfection
/
Apoptosis
/
Carcinoma, Hepatocellular
/
Proto-Oncogene Proteins c-bcl-2
/
In Situ Nick-End Labeling
/
Ethanol
Limits:
Humans
Language:
English
Journal:
Chinese Medical Journal
Year:
2002
Type:
Article
Similar
MEDLINE
...
LILACS
LIS