CAJ | 학술논문

目的:研究人钠/碘同向转运体(NIS)基因转染肺癌细胞及其蛋白表达.方法:将体外培养的肺癌A549细胞分为实验组和对照两组:以脂质体Lipofectamine2000为载体,分别介导转染重组质粒pcDNA3-hNIS和空质粒pcDNA3.NIS基因重组质粒pcDNA3-hNIS进行扩增、纯化,并经酶切鉴定和DNA测序.采用Western Blot法和免疫组化法分别检测转染肺癌细胞中NIS蛋白表达.结果:酶切鉴定年:UDNA测序结果表明重组质粒pcDNA3-hNIS中插入的hNIS基因片段大小、方向正确.Western Blot法和免疫组化染色结果显示实验组有NIS蛋白表达,其阳性率达70.6%且主要分布于细胞膜而对照组无表达.两组比较有显著性差异(P=0.000).结论:脂质体Lipofectamine 2000介导转染人钠/碘同向转运体基因pcDAN3-hNIS肺癌细胞能够成功地表达MS蛋白.
목적:연구인납/전동향전운체(NIS)기인전염폐암세포급기단백표체.방법:장체외배양적폐암A549세포분위실험조화대조량조:이지질체Lipofectamine2000위재체,분별개도전염중조질립pcDNA3-hNIS화공질립pcDNA3.NIS기인중조질립pcDNA3-hNIS진행확증、순화,병경매절감정화DNA측서.채용Western Blot법화면역조화법분별검측전염폐암세포중NIS단백표체.결과:매절감정년:UDNA측서결과표명중조질립pcDNA3-hNIS중삽입적hNIS기인편단대소、방향정학.Western Blot법화면역조화염색결과현시실험조유NIS단백표체,기양성솔체70.6%차주요분포우세포막이대조조무표체.량조비교유현저성차이(P=0.000).결론:지질체Lipofectamine 2000개도전염인납/전동향전운체기인pcDAN3-hNIS폐암세포능구성공지표체MS단백.
OBJECTIVE To examine the possibility of human sodium iodide symporter(hNIS)protein expression in lung cancer cells.MIRTHODS Human lung A549 cancer cells were thawed and cultured in vitro.The cells were divided into an experimentaI group transfected with a recombinanf pcDNA3-hNIS plasmid and a control group transfected only with a pcDNA3 plasmid.The recombinant plasmid vector encoding the hNIS gene (pcDNA3-hNIS)was amplified,purified and identified.The hNIS gene was followed by DNA sequencing.A Western blof and an immunohistochemical assay were applied 10 detect the hNIS protein expression in the transfected human lung A549 cancer cells.RESULTS Restriction enzyme digestion and DNA sequencing results showed the size and direction of the inserted gene in the recombinant pcDNA3-hNIS plasmid was correct.The Western blot method and immunohistochemical analysis showed a positive NIS protein expression in the experimental group.The NIS protein was detected mainly inthecell membranes showing a positive race up to 70.6%with no expression of fhe NIS protein in the control group.There was a significant difference between two groups (P=0.000).CONCLUSION The hNIS gene was transfected effectively into human lung A549 cancer cells mediated by Lipofectamine 2000,and was expressed with its protein in vitro.