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目的：构建多肿瘤抑制基因１（ＭＴＳ１）的真核表达载体，将其转入人源性卵巢癌细胞株ＨＯ－８９１０中，并得到稳定表达，为进一步研究其对卵巢癌细胞的影响奠定实验基础．方法：利用ＢａｍＨⅠ与ＥｃｏＲⅠ从ｐＵＣ１９－ｐ１６质粒上切下ＭＴＳ１ｃＤＮＡ片段并克隆到真核表达载体ｐｃＤＮＡ３上，构建成ＭＴＳ１真核表达载体ｐｃＤＮＡ３－ｐ１６；利用脂质体（Ｆｕ－ＧＥＮＥＴＭ６）介导的基因导入法将ｐｃＤＮＡ３－ｐ１６导入人源性卵巢癌细胞株ＨＯ－８９１０中；采用ＡＢＣ法对转染后的不同代数细胞进行免疫细胞化学检测，观察ｐ１６蛋白的表达．结果：成功构建了ＭＴＳ１真核表达载体ｐｃＤＮＡ３－ｐ１６，并进行酶切鉴定；转染后经Ｇ４１８筛选２ｗｋ出现阳性克隆８９１０－ｐ１６，并检测到其中有ｐ１６蛋白的稳定表达．结论：成功构建ＭＴＳ１真核表达载体ｐｃＤＮＡ３－ｐ１６，并可在人源性卵巢癌细胞株ＨＯ－８９１０中得到稳定表达．
OBJECTIVE: To construct a eukaryotic expression vector of multiple tumor suppressor gene 1 (MTS1) and transfer it into HO-8910 human ovarian carcinoma cell line HO-8910, and obtain stable expression, which lays the foundation for further study of its effect on ovarian cancer cells basis. METHODS: The MTS1 cDNA fragment was cut from pUC19-p16 plasmid by BamHⅠ and EcoRI and cloned into the eukaryotic expression vector pcDNA3 to construct the eukaryotic expression vector pcDNA3-p16 of MTS1. The gene mediated by Fu-GENETM6 PcDNA3-p16 was introduced into HO-8910 human ovarian cancer cell line HO-8910 by using the method of induction; the different algebraic cells after transfection were detected by immunocytochemistry using ABC method to observe the expression of p16 protein. Results: MTS1 eukaryotic expression vector pcDNA3-p16 was successfully constructed and identified by restriction enzyme digestion. Positive clones 8910-p16 were detected by G418 selection 2 h after transfection. Stable expression of p16 protein was detected. Conclusion: MTS1 eukaryotic expression vector pcDNA3-p16 was successfully constructed and stably expressed in human ovarian cancer cell line HO-8910.