[The effects of adenovirus-mediated human COX-2 antisense RNA on the growth of hepatocellular carcinoma].
OBJECTIVE
To investigate the relationship between the expression of COX-2 and liver cancer and construct a recombinant adenovirus encoding human COX-2 antisense RNA, and then to investigate its effects on liver cancer cell proliferation.


METHODS
The expression of COX-2 in 34 cases of hepatocellular carcinoma and in SMMC-7402 and SMMC-7721 cell lines was studied by using immunohistochemical techniques. The shuttle plasmid encoding anti-sense COX-2 was constructed by using cloning COX-2 cDNA fragment in the reverse direction into the pHCMVSPIA. Then the plasmid pJM17 and the shuttle plasmid were co-transferred into 293 cells with lipofectamine for homologous recombination to acquire recombinant adenovirus (Ad-AShcox-2), which was confirmed by PCR. Human hepatocellular carcinoma cell lines SMMC-7402 and SMMC-7721 were transduced in vitro. The cell apoptosis and cell cycle were analyzed by flow cytometry. The cell proliferation was determined by colony-forming efficiency.


RESULTS
We observed COX-2 expression in 82.4% of the hepatocellular carcinomas and SMMC-7402 cell line, but no COX-2 expression in the SMMC-7721 cell line. In addition, the recombinant adenovirus encoding anti-sense COX-2 fragment Ad-AShcox-2 was obtained with a titer of 1.06 x 10(12) PFU/ml. Ad-AShcox-2 reduced the expression of COX-2 and enhanced the percentage of cells into G1/G0 phase in the SMMC-7402 cell line. The difference of apoptotic index between the Ad-AShcox-2 group and the control group was statistically significant in SMMC-7402 but not in SMMC-7721. Similarly, colony-forming rates of SMMC-7402 and SMMC-7721 cell lines after Ad-AShcox-2 being transferred were (2.7+/-0.94)% and (33.6+/-4.24)%, respectively.


CONCLUSIONS
By reducing the expression of COX-2 in hepatocellular carcinoma cells with the expression of COX-2, the cells could be inhibited.
Wang
Xiao-hu
XH
Department of Gastroenterology, Taihe Hospital, Yunyang Medical College, Shiyan 442000, China.
Li
Sheng-bao
SB
Xie
Guo-jian
GJ
Yi
Jian-hua
JH
Wu
Qing-ming
QM
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis / Q000502:physiology; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D051546:Cyclooxygenase 2 / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D016372:RNA, Antisense / Q000096:biosynthesis* / Q000235:genetics
D008565:Membrane Proteins; D016372:RNA, Antisense; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human