激光杂志
激光雜誌
격광잡지
LASER JOURNAL
2013年
4期
130-131
,共2页
TDL复合物%肝癌细胞株HepG2%基因转染
TDL複閤物%肝癌細胞株HepG2%基因轉染
TDL복합물%간암세포주HepG2%기인전염
TDL compound%Hepatoma cell line HepG2%Gene transfection
目的：探讨HIV -1 Tat蛋白转导域/质粒DNA/Liposome （TDL ）复合物提高增强型绿色荧光蛋白质粒（pEGFP ）在体外培养肝癌细胞株HepG2中转染效率的可行性。方法2？l Lipofectamine 2000加入以不同电荷比混匀的Tat肽与质粒DNA溶液中制成TDL复合物。琼脂糖凝胶电泳法观察Tat肽与质粒DNA的结合力；荧光显微镜和流式细胞仪观察TDL复合物、HIV -1 Tat 肽与Lipofectamine 2000介导质粒DNA体外转染HepG2的效果；MTT法分析质粒DNA转染对HepG2活性的影响。结果琼脂糖凝胶电泳显示，各TDL复合物组均未发现明显DNA条带。TDL复合物的转染率优于单用Lipofectamine 2000（P＜0．05），HIV-1 Tat肽未发生基因转染。DNA/Tat 电荷比为1﹕8时，TDL复合物的转染效率最高。与空白组比较，各TDL复合物组、HIV -1 Tat肽组对细胞活性无明显影响（P＞0．05），Lipofectamine 2000组降低细胞活性（P＜0．05）。结论：TDL复合物可明显提高体外基因转染HepG2效率，该方法为提高非病毒载体的转染效率提供一个新策略，可能成为肝癌基因治疗的有效手段之一。
目的：探討HIV -1 Tat蛋白轉導域/質粒DNA/Liposome （TDL ）複閤物提高增彊型綠色熒光蛋白質粒（pEGFP ）在體外培養肝癌細胞株HepG2中轉染效率的可行性。方法2？l Lipofectamine 2000加入以不同電荷比混勻的Tat肽與質粒DNA溶液中製成TDL複閤物。瓊脂糖凝膠電泳法觀察Tat肽與質粒DNA的結閤力；熒光顯微鏡和流式細胞儀觀察TDL複閤物、HIV -1 Tat 肽與Lipofectamine 2000介導質粒DNA體外轉染HepG2的效果；MTT法分析質粒DNA轉染對HepG2活性的影響。結果瓊脂糖凝膠電泳顯示，各TDL複閤物組均未髮現明顯DNA條帶。TDL複閤物的轉染率優于單用Lipofectamine 2000（P＜0．05），HIV-1 Tat肽未髮生基因轉染。DNA/Tat 電荷比為1﹕8時，TDL複閤物的轉染效率最高。與空白組比較，各TDL複閤物組、HIV -1 Tat肽組對細胞活性無明顯影響（P＞0．05），Lipofectamine 2000組降低細胞活性（P＜0．05）。結論：TDL複閤物可明顯提高體外基因轉染HepG2效率，該方法為提高非病毒載體的轉染效率提供一箇新策略，可能成為肝癌基因治療的有效手段之一。
목적：탐토HIV -1 Tat단백전도역/질립DNA/Liposome （TDL ）복합물제고증강형록색형광단백질립（pEGFP ）재체외배양간암세포주HepG2중전염효솔적가행성。방법2？l Lipofectamine 2000가입이불동전하비혼균적Tat태여질립DNA용액중제성TDL복합물。경지당응효전영법관찰Tat태여질립DNA적결합력；형광현미경화류식세포의관찰TDL복합물、HIV -1 Tat 태여Lipofectamine 2000개도질립DNA체외전염HepG2적효과；MTT법분석질립DNA전염대HepG2활성적영향。결과경지당응효전영현시，각TDL복합물조균미발현명현DNA조대。TDL복합물적전염솔우우단용Lipofectamine 2000（P＜0．05），HIV-1 Tat태미발생기인전염。DNA/Tat 전하비위1﹕8시，TDL복합물적전염효솔최고。여공백조비교，각TDL복합물조、HIV -1 Tat태조대세포활성무명현영향（P＞0．05），Lipofectamine 2000조강저세포활성（P＜0．05）。결론：TDL복합물가명현제고체외기인전염HepG2효솔，해방법위제고비병독재체적전염효솔제공일개신책략，가능성위간암기인치료적유효수단지일。
Objective :To investigate the possibility of Tat/DNA/Liposome compound enhancing transfection efficiency of plasmid vector coding enhanced green fluorescence protein (pEGFP) in human hepatoma cell line HepG2 cells .Methods Tat peptide was mixed with Plasmid DNA by various charge ratio ,then added 2?l Lipofectamine 2000 to form TDL compound .Bonding force of Tat peptide and plasmid DNA was analyzed by agarose gel electrophoresis .Transfection effect of TDL compound ,HIV -1 Tat and Lipofectamine 2000 in HepG2 was observed by fluorescent microscopy and Fluorescence Activated Cell Scan .The viability of HepG2 was measured by MTT assay .Results A visible band was not found in TDL compound groups by agarose gel electrophoresis .Transfection efficiency of TDL compound was higher than that of Lipofectamine 2000 (P?0 .05) .No transfection was happened in HIV -1 Tat .When charge ratio of DNA and Tat was 1 :8 , transfection efficiency of TDL compound was the highest .There was no significant difference in cell viability between TDL groups and control group (P>0 .05) , while significant difference between Lipofectamine 2000 and control group (P?0 .05) .Conclusion TDL compound can enhance gene transfection efficiency .This method offers a new strategy to increase the transfection efficiency of non -viral genetic vector , and it may be a useful tool for gene therapy of hepatocellular carcinoma (HCC ) .
