摘要
目的探讨封闭人端粒酶RNA功能区的反义寡核苷酸(human telomerase RNA antisense oligonucleotide,hTR ASODN)对结直肠癌SW480细胞凋亡及凋亡诱导因子(apoptosis inducing factor,AIF)表达的影响。方法通过脂质体将hTR ASODN1次/d,连续3d转染入人结直肠癌SW480细胞,以未加ASODN的脂质体空白组及脂质体转染的正义组为对照。荧光显微镜观察转染情况,流式细胞仪测定转染后SW480细胞的凋亡率,RT-PCR法测定转染后AIF的表达情况。结果 hTR ASODN转染细胞后,倒置显微镜下观察各组细胞,细胞内有绿色荧光均匀分布表示转染良好。流式细胞仪测定转染24、48、72h后,各实验组凋亡率分别为:空白组(3.07±0.11)%、(3.71±0.63)%、(4.23±0.41)%,正义组(4.28±0.41)%、(4.49±0.56)%、(5.13±0.56)%,反义组(13.11±0.32)%、(22.04±0.21)%、(16.71±0.82)%,反义组分别与空白组、正义组比较,差异有统计学意义(P<0.05)。空白组、正义组、反义组光密度比值分别为(0.1527±0.0021)、(0.1580±0.0078)、(0.3683±0.0109),反义组分别与空白组、正义组比较,差异有统计学意义(P<0.05)。结论 hTR ASODN具有明显的促进凋亡、抗肿瘤作用。
Objective To explore the effects of the human telomerase RNA （hTR ） antisense oligodeoxynueleotide （ASODN） on cell apoptosis and expression of apoptosis inducing factor （AIF） of human colon cancer SW480 cells. Methods hTR antisense oligodeoxynueleotide was transfected into SW480 cells by liposome once a day with contact transfection for 3 d. The SW480 cells with blank plasmid or with ASODN sense oligodeoxynueleotide by liposome transfection served as control. The expression of green fluorescent protein （GFP） was observed by fluorescence microscopy. The cell apoptosis in SW480 cells after transfection were studied by flow cytometry. The expression of AIF was measured by RT-PCR. Results After hTR antisense oligodeoxynueleotide was transfected into SW480 cells, GFP was equally distributed in the cells. In 24, 48 and 72 h after transfection, flow cytometry indicated that the apoptotic percentages were （3.07±0.11）%, （3.71±0.63）% and （4.23±0.41）% in the blank group,（4.28±0.41）%, （4.49±0.56）% and （5.13±0.56）% in the sense group, and （13.11±0.32）%, （22.04±0.21）% and （16.71±0.82）% in anti-sense group. Significant differences were found in the anti-sense group compared with the blank group and the sense group at every time point （P0.05）, while no difference was seen between the sense group and the blank group （P0.05）. There were more AIF mRNA expression in the anti-sense group than in the blank group and the sense group （0.368 3±0.010 9 vs 0.152 7±0.002 and 0.158 0±0.007 8, P0.05）, while there was no difference between the later 2 groups （P0.05） . Conclusion hTR antisense oligodeoxynueleotide have obvious anti-tumor effect by improving apoptosis in SW480 cells..
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第22期2400-2403,共4页
Journal of Third Military Medical University
基金
重庆市卫生局医学科研项目计划(2008-2-227)~~
关键词
HTR
反义寡核苷酸
结直肠癌
凋亡
凋亡诱导因子
hTR
antisense oligodeoxynueleotide
colon cancer
apoptosis
apoptosis inducing factor
