ABCE1基因对人食管癌细胞Eca109的影响及作用机制  

Effect of ABCE1 gene on human esophageal cancer cells Eca109 and its mechanism

作　　者：黄波[1] 龚向南[1] 周红丽[2] 王思望[1] 熊飞[1] 

机构地区：[1]辽宁医学院附属第一医院胸外科,辽宁锦州121000 [2]辽宁医学院附属第一医院肾内科,辽宁锦州121000

出　　处：《解放军医学院学报》2014年第9期942-946,共5页Academic Journal of Chinese PLA Medical School

基　　金：辽宁省科技厅课题(201202143;2013022038)~~

摘　　要：目的明确ABCE1基因对食管癌Eca109细胞中的增殖、侵袭、迁移、凋亡生物学行为的影响并对其作用机制进行初步探讨。方法将构建好的ABCE1的SiRNA绿色荧光载体转染入食管癌Eca109细胞中,于荧光显微镜下观测转染效率并应用Western blot法及RT-PCR法证实基因沉默的效果,通过Western blot法检测RNase L的改变,MTT法分析细胞增殖能力并绘制细胞生长曲线,Transwell法检测细胞侵袭力,Transwell法检测细胞侵袭力,流式细胞术检测细胞凋亡率。结果转染ABCE1-SiRNA后的Eca109细胞内可见绿色荧光。Western blot法及RT-PCR法证实了基因沉默的效果(P<0.05)。MTT实验证实,转染ABCE1-SiRNA载体的食管癌细胞与阴性对照组和空白对照组的细胞相比,增殖受到显著抑制(P<0.05)。Transwell结果显示转染ABCE1-SiRNA载体的食管癌细胞穿膜细胞数较阴性对照组和空白对照组的细胞明显减少(P<0.05)。转染ABCE1-SiRNA-1和转染ABCE1-SiRNA-2的细胞与空白对照组细胞和转染ABCE1-SiRNA-N的细胞相比较,细胞迁移发生明显减慢(P<0.05),流式细胞仪检测细胞凋亡显示转染ABCE1-SiRNA载体的细胞的细胞凋亡率均显著高于阴性对照组和空白对照组细胞(P<0.05)。结论抑制ABCE1基因表达后,食管癌细胞的增殖、侵袭及迁移能力被抑制,细胞发生早期凋亡。这表明ABCE1基因的表达对食管癌细胞Eca109具有肿瘤生物学行为的影响。Objective To study the effect of ABCE1 gene on proliferation, invasion, migration and apoptosis of human esophageal cancer cells Eca109 and its mechanism. Methods The ABCE1-SiRNA green fluorescence vector was constructed and transfected into Eca109. The transfection efficiency was observed under a fluorescence microscope. The gene silencing effect was detected by Western blot and RT-PCR, respectively. The changed RNaseL was detected by Western blot. The cell proliferation was analyzed by MTT and its growth curve was plotted. The cell invasion was detected by transwell, and the cell apoptosis was assayed by flow cytometry. Results Green fluorescence was observed in the ABCE1-SiRNA-transfected Eca109. The gene silencing effect was confirmed by Western blot and RT-PCR, respectively （P 〈 0.05）. MTT showed that the proliferation of ABCE1-SiRNA-transfected Eca109 was significantly lower than that of negative and blank control cells （P 〈 0.05）. Transwell revealed that the number of membrane- penetrating Eca 109 was significantly less in ABCE 1-SiRNA-transfected vector than in negative and blank control cells （P 〈 0.05）. The migration of ABCE1-SiRNA-1 and 2-transfected Ecal09 was obviously slower than that of negative and blank control cells and ABCEI-SiRNA-N- transfected Eca109 （P 〈 0.05）. Flow cytometry showed that the apoptosis rate of ABCE1- siRNA- transfected Eca109 was significantly higher than that of negative and blank control cells （P 〈 0.05）. Conclusion The proliferation and invasion of ABCE1-SiRNA-transfected Eca109 are inhibited with early apoptosis occurred, suggesting that ABCEl expression can affect the biological behavior of Eca109.

关 键 词：ECA109细胞 三磷酸腺苷结合盒转运子E1基因 细胞增殖 细胞凋亡 

分 类 号：R735.1[医药卫生—肿瘤]