中文摘要：

目的构建CD59第40位氨基酸突变的重组体，研究Hela细胞中CD59基因突变引起肿瘤逃逸相关分子caspase-3表达量的变化。方法采用重组聚合酶链反应定点诱变技术构建第40位氨基酸突变的CD59，克隆人pALTER-MAX质粒，采用阳离子脂质体法将重组质粒转染Hela细胞。G418筛选稳定表达细胞克隆，免疫酶标、免疫荧光、ELISA筛选出高表达CD59的细胞株，用免疫组化法检测转染前后Hela细胞内caspase-3表达量的变化。结果①酶切鉴定和序列测定均证实成功构建了CD59第40位氨基酸突变的重组质粒。②转染突变CD59后Hela细胞内caspase-3的表达明显增加，与未转染的Hela细胞比较（P〈0．01）差别有显著差异。结论caspase-3的表达变化可能是CD59引起肿瘤逃逸的另一途径。

英文摘要：

Objective To construct mutant CD59 molecule （mutated at the 40th amino acid） and study the alleosis in the expression of caspase-3 caused by the mutant CD59 in Hela cells. Methods Mutant CD59 DNA was inserted into the vector pALTER-MAX and transfected into the Hela cells via lipofectamine method. Stable expression clones were selected by the addition of G418. The G418-resistant clones, which expressed relatively high levels of mutant CD59, were sorted by immunofluorescence method, enzyme immunohistochemistry, and ELISA. The expression of caspase-3 in Hela cells before and after transfection was tested by immunohistochemistry. Results （1）Recombinant plasmids of pALTER-MAX-CD59 which was mutated at the 40th amino acid had been successfully constructed according to thesequence of enzyme digestion analysis and fragment investigation. （2）The amount of caspase-3 in transfected Hela cells was obviously higher than that in nomlal （ P 〈 0.01 ）. Conclusion The other way by which CD59 lead the tumor escaping may be realized through the expression changing of caspase-3.