华中科技大学学报（医学版）
華中科技大學學報（醫學版）
화중과기대학학보（의학판）
ACTA UNIVERSITATIS MEDICINAE TONGJI
2009年
5期
598-601
,共4页
RNA%小分子干扰%PLAGL2%SP-C%SP-B%H441细胞
RNA%小分子榦擾%PLAGL2%SP-C%SP-B%H441細胞
RNA%소분자간우%PLAGL2%SP-C%SP-B%H441세포
siRNA,PLAGL2%SP-C,SP-B,H441 cells
目的 构建PLAGL2 siRNA表达载体质粒并探讨其体外干扰作用.方法 构建的PLAGL2 siRNA表达载体质粒经脂质体介导转染到小鼠肺Ⅱ型上皮细胞株H441细胞中并获得稳定细胞株,采用实时荧光PCR及Western blot技术分别检测转染组和野生型H441细胞中PLAGL2、SP-C、SP-B mRNA和PLAGL2蛋白表达水平的变化.结果 实时荧光PCR检测结果显示:与野生型H441细胞相比较,PLAGL2 siRNA表达载体质粒转染的H441细胞中PLA-GL2 mRNA水平明显降低(P<0.05),SP-C mRNA水平明显增高(P<0.05),而SP-B mRNA水平没有明显差异(P>0.05);Western blot检测结果显示:与野生型H441细胞相比较,PLAGL2 siRNA表达载体质粒转染的H441细胞中PLAGL2蛋白表达水平明显降低(P<0.05).结论 成功构建了PLAGL2 siRNA表达载体,将其转染至H441细胞中可有效抑制PLAGL2基因的表达,同时促进SP-C基因的表达.
目的 構建PLAGL2 siRNA錶達載體質粒併探討其體外榦擾作用.方法 構建的PLAGL2 siRNA錶達載體質粒經脂質體介導轉染到小鼠肺Ⅱ型上皮細胞株H441細胞中併穫得穩定細胞株,採用實時熒光PCR及Western blot技術分彆檢測轉染組和野生型H441細胞中PLAGL2、SP-C、SP-B mRNA和PLAGL2蛋白錶達水平的變化.結果 實時熒光PCR檢測結果顯示:與野生型H441細胞相比較,PLAGL2 siRNA錶達載體質粒轉染的H441細胞中PLA-GL2 mRNA水平明顯降低(P<0.05),SP-C mRNA水平明顯增高(P<0.05),而SP-B mRNA水平沒有明顯差異(P>0.05);Western blot檢測結果顯示:與野生型H441細胞相比較,PLAGL2 siRNA錶達載體質粒轉染的H441細胞中PLAGL2蛋白錶達水平明顯降低(P<0.05).結論 成功構建瞭PLAGL2 siRNA錶達載體,將其轉染至H441細胞中可有效抑製PLAGL2基因的錶達,同時促進SP-C基因的錶達.
목적 구건PLAGL2 siRNA표체재체질립병탐토기체외간우작용.방법 구건적PLAGL2 siRNA표체재체질립경지질체개도전염도소서폐Ⅱ형상피세포주H441세포중병획득은정세포주,채용실시형광PCR급Western blot기술분별검측전염조화야생형H441세포중PLAGL2、SP-C、SP-B mRNA화PLAGL2단백표체수평적변화.결과 실시형광PCR검측결과현시:여야생형H441세포상비교,PLAGL2 siRNA표체재체질립전염적H441세포중PLA-GL2 mRNA수평명현강저(P<0.05),SP-C mRNA수평명현증고(P<0.05),이SP-B mRNA수평몰유명현차이(P>0.05);Western blot검측결과현시:여야생형H441세포상비교,PLAGL2 siRNA표체재체질립전염적H441세포중PLAGL2단백표체수평명현강저(P<0.05).결론 성공구건료PLAGL2 siRNA표체재체,장기전염지H441세포중가유효억제PLAGL2기인적표체,동시촉진SP-C기인적표체.
Objective To construct human PLAGL2 siRNA expression DNA plasmid and study the interfering effect on gene expression in H441 cells. Methods H441 cells were transfected with PLAGL2 siRNA expression DNA plasmid with Fu-gene6. Real time PCR and Western blot were employed to detect the interfering effect on the expression of PLAGL2.SP-CSP-B mRNA and protein in wildtype and PLAGL2 siRNA expression DNA plasmid transfected H441 cells respectively. Results Real time PCR revealed that,as compared with wildtype H441 cells, the expression level of PLAGL2 mRNA in PLAGL2 siRNA plasmid transfected H441 cells was significantly reduced(P<0. 05) ,but that of SP-C mRNA in PLAGL2 siRNA plasmid transfected H441 cells was significantly increased(P<0. 05). Western blot showed that,as compared with wildtype H441 cells, the expression level of PLAGL2 protein in PLAGL2 siRNA DNA plasmid transfected H441 cells was significantly reduced(P< 0. 05). Conclusion Human PLAGL2 siRNA expression DNA plasmid was constructed successfully,and its transfection into the H441 cells could effectively inhibit the PLAGL2 expression,and simultaneously promote the expression of SP-C.
