缺氧诱导因子-1α反义寡核苷酸体外对胶质瘤U87细胞增殖和侵袭的影响  

Effects of antisense oligodeoxynucleotides targeting hypoxia inducible factor 1α gene on huinan glioma cell line U87 in vitro

作　　者：李巧玉[1] 张焱[1] 袁志诚[1] 陆培松[1] 湛利平[1] 杨勇[1] 

机构地区：[1]江苏大学附属人民医院神经外科,镇江212002

出　　处：《中华实验外科杂志》2011年第1期89-91,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察缺氧诱导因子-1α（HIF-1α）反义寡核苷酸体外对人胶质瘤U87细胞增殖、凋亡和侵袋的影响。方法人工合成的HIF-1α反义短发夹经阳离子脂质体包裹后瞬时转染人胶质瘤细胞株U87，采用Western blot法检测转染后胶质瘤细胞HIF-1α蛋白表达，证实转染成功，再采用噻唑蓝（MTF）比色法和Transwell小室体外侵袭实验检测转染后对U87细胞增殖体外侵袭能力的影响，流式细胞仪检测细胞凋亡。结果转染靶向HIF-1α的短链发夹RNA的胶质瘤细胞，HIF-1α蛋白表达较空白细胞组明显降低，MTT法检测转染组、空载体组和空细胞组24h细胞的增殖率分别为5．46％、21．25％、22．32％，体外侵袭性实验表明转染组、空载体组和空细胞组细胞12h侵袭ECM的细胞数分别为（22±4）、（124±3）、（122±6）；流式细胞仪检测表明转染组、空载体组和空细胞组细胞凋亡率分别为（53．35±2．80）％、（12．02±1．60）％、（10．19±3．15）％，转染组与空白细胞组及空载体组比较差异均有统计学意义（P〈0．05）。结论封闭HIF-1α蛋白的表达，可以抑制人胶质瘤U87细胞增殖和侵袭能力，促进细胞捌亡。Objective To investigate the effects of antisense oligodeoxynucleotides （ODNs） targe- ting hypoxia inducible factor-1α（HIF-1α） on the apoptosis, proliferation and invasion of U87 glioma cell line. Methods Antisense ODNs were constructed and transfected into U87 cells by Dosper liposomal reagent. The HIF-1α gene expression was detected by Western blotting, the cell proliferative index was determined by methyl thiazol tetrazolium （MTT） assay, the cells cycle and apoptosis of the cells were examined by flow cytometry and the changes of the U87 cells invasive ability were measured by Transwell chamber. Results The protein expression of HIF-1α in U87 ceils was down-regulated by HIF-1α ASONDN. The cell proliferative index in transfected group, empty vector group and control group was 5.46%, 21.25% and 22. 32% respectively. Transwell chamber assay showed that the cell number in transfected group, empty vector group and control group was （22 ±4）, （124±3） and （122 ±6） respectively; and the apoptosis rate was （53.35 ± 2.80） %, （ 12.02 ± 1.60） %, （ 10. 19 ± 3. 15 ） % respectively, and there was significant difference between transfected group and other groups （ P 〈 0. 05）. Conclusion Silencing the expression of HIF-1α protein can inhibit proliferation and invasion, and promote apoptosis of human glioma U87 cells. HIF-1α is expected to become a target for cancer therapy.

关 键 词：胶质瘤 缺氧诱导因子-1Α 侵袭 增殖 

分 类 号：R739.4[医药卫生—肿瘤]