构建表达人环氧化酶－2（cyclooxygenase-2,cox-2)反义RNA的腺病毒载体，研究其对人食管癌细胞生长的影响。方法：把人cox-2基因的cDNA片段反向克隆于穿梭质粒pHCMVSP1A的CMV启动子之下，获得pAd-AShcox-2，通过脂质体与pJM17共转染293细胞，经同源重组产生编码cox-2反义RNA的重组腺病毒－Ad-AShcox-2，PCR鉴定为阳性克隆并大量扩增，转染食管癌细胞EC9706，用生长细胞计数，3H－TdR掺入及免疫细胞化学的方法，研究对食管癌细胞生长，cox-2表达，DNA合成的影响，结果：成功构建编码cox-2反义RNA的重组腺病毒Ad-AShcox-2，病毒感染EC9706后，cox-2表达降低，3H－TdR掺入量减少，与对照组比较P＜0．001，同时发现食管癌细胞的生长受抑制，结论：减少cox-2的表达可能是治疗食管癌的一种新途径。

To construct the recombinant adenovirus encoding human cox-2 antisense RNA and to investigate its effects on proliferation of tumor cells. 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 homologous recombinantion to acquire recombinant adenovirus which was confirmed by PCR . We evaluated how alterations of cox-2 expression in EC9706 cells transfected with adenovirus-mediated human cox-2 antisense RNA expression (Ad-AShcox-2) or adenovirus recombinants carrying LacZ gene (Ad-LacZ), and its effects on cell proliferation were determined by cell growth rate, 3 H-TdR incorporation and immunohistochemistry. Results: The recombinant adenovirus encoding antisense cox-2 fragment Ad-AShcox-2 was obtained with the titer of 0.86 10 12 PFU/ml. Ad-AShcox-2 could reduce the expression of cox-2 , and strongly inhibit cell growth rate and cause cellular death. Simultaneously, 3 H-TdR incorporation was lower than that of control group ( P <0.001) Conclusion: Reduction of cox-2 expression could inhibit esophageal cancer proliferation through inhibiting the synthesis of DNA.

郧阳医学院基金