脂质体介导外源基因转入人晶体上皮细胞的实验研究

目的确定外源基因能否由脂质体携带进入人原代晶体上皮细胞（ｐｒｉｍａｒｙｈｕｍａｎｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｓ,ＰＨＬＥＣｓ）。方法体外培养人原代晶体上皮细胞,用阳离子脂质体（ｌｉｐｏｆｅｃｔｉｎｒｅａｇｅｎｔ,ＬＲ）携带重组质粒报道基因（β半乳糖苷酶）,向人原代晶体上皮细胞转移,在转移１２、２４、３６小时,分别表达２天和６天时,用以Ｘｇａｌ为底物的酶显色方法,检测报道基因对人原代晶体上皮细胞的转移率。结果脂质体（ｌｉｐｏｆｅｃｔｉｎ）介导的外源基因可以转入人原代晶体上皮细胞,在转移２４小时,表达２天时转移率可达４８％。结论稳定的外源基因可由脂质体介导转入生长中人原代晶体上皮细胞,作为晶体上皮细胞基因类药物介入的基础研究和应用研究的介导体,脂质体显示出有希望的前景。借此方法也可以从外源基因入手,对晶体上皮细胞的生理和病理活动,进行机制研究。

Experimental observation of foreign gene transfer with lipofectin to the primary human lens epithelial cells

OBJECTIVE: To determine whether an exogenous gene carried by lipofectin can be introduced into the primary human lens epithelial cells. METHODS: Plasmid DNA with beta-galactosidase gene carried by lipofectin was applied to primary cultured human lens epithelial cells. Gene expression was detected by enzymatic color reaction using X-gal as a substrate in the 2 and 6 days of expression after 12, 24, 36 hours of transfection respectively. The gene transfer positive rate of the lens epithelial cells was counted. RESULTS: The foreign gene could be transferred into primary human lens epithelial cells by lipofectin. In the expression of 2 days, transfer positive rate was up to 48% after 24 hours of transfection. CONCLUSIONS: Efficient and stable transfer of the functional gene can be achieved by lipofectin into the lens epithelial cells. Lipofectin is an available and a promising vehicle for delivering aim gene and studying on the mechanism of physiology and pathology of lens epithelial cells.