人乙酰胆碱酯酶基因转染增强全反式维甲酸诱导胰腺癌细胞凋亡  

The transfer of human acetylcholinesterase gene to pancreatic cancer cells enhances the effect of all-trans retinoic acid induced cell apoptosis

作　　者：夏璐[1] 乔敏敏[1] 章永平[1] 袁耀宗[1] 金其煌[2] 晋鹏[2] 张学军[2] 

机构地区：[1]上海第二医科大学附属瑞金医院消化科,200025 [2]中国科学院生化与细胞生物学研究所

出　　处：《中华消化杂志》2005年第5期287-290,共4页Chinese Journal of Digestion

摘　　要：目的探讨转染人乙酰胆碱酯酶(AChE)基因对全反式维甲酸诱导胰腺癌细胞凋亡的影响。方法应用含人AChE基因的真核表达质粒pcDNA3/AChE和绿色荧光蛋白(GFP)真核表达质粒pcDNA3/GFP,以脂质体介导上述两种质粒DNA对人胰腺癌细胞株Patu8988进行瞬时转染,转染后的细胞经全反式维甲酸(ATRA,1μmol/L)分别处理24h及48h,流式细胞仪分别检测AnnexinV染色后早期细胞凋亡率及转染效率,以细胞化学染色法(KarnovskyRoots染色)检测AChE表达。结果流式细胞仪测定GFP显示转染效率达(45.50±3.84)%,人AChE基因转染后联合ATRA治疗凋亡比例增加,转染前后分别为(14.26±1.16)%/(42.84±7.21)%(24h,P<0.01)和(18.08±0.75)%/(56.29±2.54)%(48h,P<0.01)。细胞化学染色法可见AChE阳性细胞,染色聚集于胞核部位。结论胰腺癌细胞AChE基因的高表达可增加细胞对ATRA诱导凋亡的敏感性。Objective To study the effect of transferred apoptosis-associated acetylcholinesterase (AChE) gene on all-trans retinoic acid (ATRA) induced pancreatic cancer cell apoptosis. Methods The eukaryotic expression vector containing human AChE(pcDNA/AChE) and green fluorescence protein(GFP)(pcDNA3/GFP)were transiently transfected into Patu 8988 cells,a human pancreatic cancer cell line,mediated by liposome Lipofectamine TM.After transfection,cells were treated with ATRA for 24 h and 48 h.The expression of GFP in the cells was examined using fluorescence microscope every day after transfection.Flow cytometry (FCM) (Annexin V staining) and cytochemical staining(Karnovsky-Roots method) were used to determine apoptosis rate and AChE expression,respectively. Results The positive GFP expression rate in Patu 8988 cells 24 h after transfection was (45.50±3.84)% . After being treated with ATRA, the cell apoptotic rates detected by FCM were (42.84±7.21)%(24 h)and (56.29[JP2] ±2.54)%(48 h)respectively,higher than that in untreated group[24 h:(14.26±1.16)%,P<0.01[JP];48 h:(18.08±0.75)%,P<0.01]. AChE positive cells could be seen by using cytochemical staining, with a pattern of nuclei accumulation of this staining. Conclusions Overexpression of AChE gene could increase the ATRA-induced apoptosis in pancreatic cancer cells, but it is unclear how AChE contributes to this apoptotic process.

关 键 词：乙酰胆碱酯酶 癌细胞凋亡 维甲酸诱导 基因转染 细胞化学染色法 PCDNA3 真核表达质粒 人胰腺癌细胞株 Annexin 流式细胞仪 AChE 绿色荧光蛋白 全反式维甲酸 转染效率 ATRA 质粒DNA 脂质体介导 细胞凋亡率 方法应用 瞬时转染 

分 类 号：R735.9[医药卫生—肿瘤]