Effects of antisense oligonucleotides of PKC-alpha on proliferation and apoptosis of HepG2 in vitro.
BACKGROUND
Hepatocellular carcinoma (HCC) is one of the most common malignancies in China. The long-term survival rate of patients with HCC after prevention and management remains unsatisfactory. In order to provide a novel strategy to cure HCC, we investigated the effects of antisense oligonucleotides of PKC-alpha on proliferation and apoptosis of human hepatoma cell line HepG2 in vitro.


METHODS
The human hepatocellular carcinoma cell line HepG2 was cultured and subcultured in RPMI1640 medium in vitro. PKC-alpha antisense oligonucleotides(asODN) of different concentrations with a random sequence as a control were transfected into HepG2 cells by lipofectin(LP). The cell growth index (GI) and the clone formation rate of HepG2 were detected by MTT colorimetric assay and soft agar assay, respectively. The apoptosis rate of HepG2 treated with PKC-alpha asODN was assayed by flow cytometry(FCM). The results were analyzed by SPSS 10.0 software.


RESULTS
The GI of HepG2 transfected by PKC-alpha asODN with concentrations ranging from 0.10 micromol to 1.00 micromol were lower significantly than those of control groups (P<0.05). The clone formation rates of HepG2 transfected by PKC-alpha asODN from 0.05 micromol to 1.00 micromol were lower significantly than those of the control groups (P<0.01), and there was a dose-dependent relationship among them. The apoptosis rates of HepG2 treated with PKC-alpha asODN from 0.50 micromol to 1.00 micromol were significantly higher than those of the control groups.


CONCLUSIONS
PKC-alpha asODN could inhibit the growth and proliferation of HepG2 and induce its apoptosis by blocking the cell signal transduction related to PKC-alpha in vitro, and may be potentially used in the prevention and management of recurrent and metastatic HCC.
Zhu
Bao-Hua
BH
Department of Hepatobiliary Surgery, First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400016, China. baohuazhu@sohu.com
Yao
Zhen-Xiang
ZX
Luo
Shao-Jun
SJ
Jiang
Li-Ming
LM
Xiao
Jiang-Wei
JW
Liu
Sheng-Chun
SC
Liu
Jin-Biao
JB
Sun
Jian-Ming
JM
Pei
Zhi-Yuan
ZY
D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D008297:Male; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D011336:Probability; D011493:Protein Kinase C / Q000494:pharmacology*; D051571:Protein Kinase C-alpha; D012680:Sensitivity and Specificity
D016376:Oligonucleotides, Antisense; C496628:PRKCA protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha