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Sheng-Bao Li, Qing-Ming Wu, Qiang Wang, Xiao-Hu Wang, Guo-JianXie, Department of Gastroenterology, Taihe affiliated Hospital, Yunyang Medical college, Shiyan 442000, Hubei Province, China
Correspondence to: Dr. Sheng-Bao Li, Department of Gastroenterology, Taihe Affiliated Hospital, Yunyang Medical College, 29 Renmin south Road, Shiyan 442000, Hubei Province, China. libao2000@ 163.com
Received: August 10, 2002 Revised: August 15, 2002 Accepted: August 23, 2002 Published online: May 15, 2003
AIM
To construct the recombinant adenovirus encoding human cox-2 antisense RNA, and to investigate its effect on synthesis of DNA and proteins in esophgeal carcinoma cell line EC9706.
METHODS
The shuttle plasmid encoding antisense cox-2 was constructed by cloning cox-2 cDNA fragment in the reverse direction into the pHCMVSP1A. Then the plasmid pJM17 and the shuttle plasmid were cotransferred into 293 cells with lipofectamine for homologouserecombinantion to acquire recombinant adenovirus confirmed by PCR. The expressions of cox-2 in esophgeal carcinoma cell line EC9706 cells were evaluated, and its effects on cell proliferation were determined by cell growth rate, 3H-TdR and 3H-Leucine incorporation.
RESULTS
The recombinant adenovirus encoding antisense cox-2 fragment ad-AShcox-2 was obtained with the titer of 0.86±1012 PFU/ml. Ad-AShcox-2 can reduce the expression of cox-2, and inhibit cell growth rate and cause cellular death. Meanwhile, The efficiency of 3H-TdR and 3H-Leucine incorporation was significant lower than that in the control group at 48, 72, 96 hours (q48 h = 16.36 vs 16.36, q72 h = 39.07 vs 19.90 , q96 h= 54.80 vs 30.33; P<0.001).
CONCLUSION
Reducing the expression of cox-2 may inhibit the proliferation of esophageal cancer cells through inhibiting the synthesis of DNA and protein.
Key Words: N/A
Citation: Li SB, Wu QM, Wang Q, Wang XH, Guo-JianXie. Effects of adenovirus-mediated human cox-2 antisense RNA on synthesis of DNA and proteins in esophgeal carcinoma cell line. Shijie Huaren Xiaohua Zazhi 2003; 11(5): 517-521
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