[Inhibitory effect of adriamycin combined P21CIP1 gene transfection on proliferation of human hepatoblastoma cell line HepG2].
OBJECTIVE
Chemotherapy protocols using adriamycin (ADM) is a standard treatment for hepatoblastoma, but the treatment results became unsatisfied because of drug resistance. Recently, ADM combined gene therapy is a developing alternative treatment for hepatoblastoma. This study was to investigate the effect of ADM combined human p21CIP1 transfection on the proliferation of hepatoblastoma cell line HepG2.


METHODS
HepG2 cells were divided into empty control group (no treatment), ADM group (treated with 0.5 microg/mL ADM), blank control group (transfected with blank plasmid pcDNA3), p21 group (transfected with plasmid pcDNA3-p21), and combination group (ADM treatment plus p21 transfection). The proliferation of HepG2 cells was observed by MTT assay. The mRNA levels of p21 and survivin were detected by real-time polymerase chain reaction (PCR).


RESULTS
After transfection, the mRNA level of p21 in p21 group was increased by 155 folds of that in empty control group (P<0.05). p21 inhibited the proliferation of HepG2 cells at Day 3 and Day 4 after transfection (P<0.01). The proliferation inhibition rate was significantly higher in combination group than in ADM group and p21 group (43.92% vs. 32.97% and 35.77% at Day 3, P<0.01; 59.86% vs. 39.35% and 40.96% at Day 4, P<0.01; 51.81% vs. 33.91% and 10.68% at Day 5, P<0.01). This effect was enhanced along with the increasing time of co-treatment from Day 1 to Day 4 (r=0.91, P<0.05), and it was obvious at Day 4 (Q =1.07). The mRNA level of survivin was significantly lower in combination group than in p21 group and ADM group (P<0.01).


CONCLUSIONS
p21 gene transfection plus ADM can inhibit the proliferation of HepG2 cells and down-regulate the level of survivin mRNA, thus may be a potential therapeutic strategy against human hepatoblastoma.
Xiong
Juan
J
Institute of Immunology,Union Hospital, Tongji Medical College,Huazhong University of Science and Technology, Wuhan, Hubei, 430022, People's Republic of China.
Hu
Li-Hua
LH
Li
Yi-Rong
YR
Wang
Lin
L
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics / Q000378:metabolism*; D004317:Doxorubicin / Q000494:pharmacology*; D056945:Hep G2 Cells; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D012333:RNA, Messenger / Q000378:metabolism; D000077022:Survivin; D014162:Transfection
D000903:Antibiotics, Antineoplastic; C107489:BIRC5 protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; D000077022:Survivin; D004317:Doxorubicin