中文摘要：

目的确定重组腺病毒p53（rAd—p53）对口腔黏膜异常增生角化细胞POE-9n转染的最佳滴度及其转染效率，为其在口腔癌前病变领域的进～步研究提供实验依据。方法采用携带绿色荧光蛋白（GFP）报告分子的重组腺病毒颗粒rAd—GFP确定口腔异常增生细胞对腺病毒转染的敏感性；以不同滴度的rAd—p53转染口腔异常增生细胞并通过MTT比色检测各组细胞的增殖抑制率；免疫细胞化学染色证实，rAd—p53转入细胞内表达P53蛋白。结果rAd—GFP转染POE-9n细胞在100～500的感染倍数（MOI）范围内转染效率都大于95％，当MOI为100、200和500时，转染效率升高不明显（r=0．124，P〉0．05）；在100～500的MOI范围rAd-p53对POE-9n细胞增殖的抑制效果显著，MOI=100、MOI=200及MOI=500组的生长抑制率在96h和120h时间点差异无统计学意义（P〉0．05）；rAd—p53转染后的POE-9n细胞中均呈现P53蛋白的强阳性表达。结论MOI=100可作为rAd—p53对POE-9n细胞转染的理想滴度，能获得较高且稳定的转染效率。

英文摘要：

Objective To investigate and evaluate the appropriate virus titer and transfection efficiency of recombinant adenovirus p53 into the oral dysplastic epithelial cells（POE-9n） and provide reference for oral precancerosis research. Methods The transfection sensitivity of adenovirus into oral dysplastic epithelial cells was evaluated by the recombinant adenovirus p53 containing green fluorescent protein （rAd-GFP）. Different titre rAd-p53 was transfected into oral dysplastic epithelial cells to evaluate the effects of rAd-p53 on cell proliferation inhibition by MTT assay. The expression of exogenous p53 genc in POE-9n cells was detected by immunoeytoehemistry. Results More than 95% POE-9n cells were transfected by rAd-GFP with MOI from 100 to 500 and there was no statistical difference between different MOI values（r=0.124, P〉0.05）. It was found that rAd-p53 had significant inhibition effects on POE- 9n cell proliferation with MOI from 100 to 500, and there were no significant differences at 96 h and 120 h after the transfection on cell proliferation inhibition（P〉0.05）. P53 protein was well expressed in rAd-p53 transfected POE- 9n cells. Conclusion Exogenous p53 can be successfully transfected into POE-9n cells by rAd-p53 and the virus titer of MOI 100 was high enough to ensure efficient transfection.