[The effects of human 21.5 kDa MBP gene on HepG-2 proliferation and apoptosis].
OBJECTIVE
To test the effects of the 21.5 kDa human brain myelin basic protein (MBP) on the proliferation and apoptosis of HepG-2.


METHODS
pSVCEP-MBP-CAT plasmid containing the full-length 21.5 kDa MBP cDNA was transfected into human hepatoma carcinoma cells (HepG-2). The pSVCEP-CAT vector transfected HepG-2 cells served as negative control. RT-PCR and Western blot were performed to confirm the effectiveness of the transfection. MTT measures were used to determine the proliferative curve of cells. H2O2 was then added to induce cell apoptosis. DNA ladder, immunohistochemistry assay, comet electrophoresis and TUNEL (Terminal Deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling) were used to detect the apoptosis and relevant protein expressions.


RESULTS
The 21.5 KDa MBP cDNA was transfected into HepG-2 cells successfully. MTT measures of pSVCEP-MBP-CAT transfected group showed increased proliferation and anti-apoptosis. The control group displayed with more typical DNA ladders and much higher level of Caspase-3 than the MBP group. Comet and TUNEL assays revealed that the control group cells had significant DNA damages and serious apoptosis, whereas the MBP group showed slight changes.


CONCLUSIONS
The 21.5 kDa MBP promotes the proliferation of HepG-2 and blocks apoptosis.
Pan
Hong
H
Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China.
Ren
Si-Chong
SC
He
Xuan-Lin
XL
Wei
Yan-Xia
YX
Wang
Ruo-Han
RH
Yang
Yin
Y
Liu
Ji
J
D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004676:Myelin Basic Protein; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
C499048:MBP protein, human; D004676:Myelin Basic Protein; D009419:Nerve Tissue Proteins; D014157:Transcription Factors