Suppression of human hepatoma (HepG2) cell growth by nuclear factor-kappaB/p65 specific siRNA.
Nuclear factor-kappaB (NF-κB) is a transcription factor and antagonist of apoptosis during liver regeneration and closely related to the formation and development of hepatocellular carcinoma. In the present study, we investigated the effect of small interference RNA (siRNA)-mediated inhibition of NF-κB on growth of human hepatoma (HepG2) cells. Our data indicated that the expression of NF-κB/p65 mRNA was significantly higher in the HepG2 cells than that in the normal liver (LO2) cells before transfection, and the expression of NF-κB/p65 in the HepG2 cells with NF-κB/p65 siRNA (100 nMol/L) transfection at 72 h was reduced at the levels of mRNA (93%) and protein (62%) using real-time reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting. Interestingly, the apoptosis index of the HepG2 cells increased up to 85%, detected by Annexin V-fluorescein isothiocyanate, suggesting that NF-κB is overexpressed in hepatoma cells and can be inhibited by NF-κB/p65 siRNA through the apoptotic mechanism. Thus, we conclude that NF-κB is a potential molecular target for HCC gene therapy.
Wu
Wei
W
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, 20 West Temple Road, Nantong 226001, Jiangsu Province, China.
Yao
Dengfu
D
Wang
Yilang
Y
Qiu
Liwei
L
Sai
Wenli
W
Yang
Junling
J
Yao
Ninghua
N
Li
Shanshan
S
Bian
Yinzhu
Y
Wang
Zhiwei
Z
Yao
Dengbing
D
D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology*; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology*; D013997:Time Factors; D014162:Transfection
D016328:NF-kappa B; D012333:RNA, Messenger; D034741:RNA, Small Interfering