中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
1期
89-91
,共3页
李巧玉%张焱%袁志诚%陆培松%湛利平%杨勇
李巧玉%張焱%袁誌誠%陸培鬆%湛利平%楊勇
리교옥%장염%원지성%륙배송%담리평%양용
胶质瘤%缺氧诱导因子-1α%侵袭%增殖
膠質瘤%缺氧誘導因子-1α%侵襲%增殖
효질류%결양유도인자-1α%침습%증식
Glioma%HIF-1α%Invasion%Proliferation
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、凋亡和侵袭的影响.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(MTT)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24 h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12 h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,促进细胞凋亡.
目的 觀察缺氧誘導因子-1α(HIF-1α)反義寡覈苷痠體外對人膠質瘤U87細胞增殖、凋亡和侵襲的影響.方法 人工閤成的HIF-1α反義短髮夾經暘離子脂質體包裹後瞬時轉染人膠質瘤細胞株U87,採用Western blot法檢測轉染後膠質瘤細胞HIF-1α蛋白錶達,證實轉染成功,再採用噻唑藍(MTT)比色法和Transwell小室體外侵襲實驗檢測轉染後對U87細胞增殖體外侵襲能力的影響,流式細胞儀檢測細胞凋亡.結果 轉染靶嚮HIF-1α的短鏈髮夾RNA的膠質瘤細胞,HIF-1α蛋白錶達較空白細胞組明顯降低,MTT法檢測轉染組、空載體組和空細胞組24 h細胞的增殖率分彆為5.46%、21.25%、22.32%,體外侵襲性實驗錶明轉染組、空載體組和空細胞組細胞12 h侵襲ECM的細胞數分彆為(22±4)、(124±3)、(122±6);流式細胞儀檢測錶明轉染組、空載體組和空細胞組細胞凋亡率分彆為(53.35±2.80)%、(12.02±1.60)%、(10.19±3.15)%,轉染組與空白細胞組及空載體組比較差異均有統計學意義(P＜0.05).結論 封閉HIF-1α蛋白的錶達,可以抑製人膠質瘤U87細胞增殖和侵襲能力,促進細胞凋亡.
목적 관찰결양유도인자-1α(HIF-1α)반의과핵감산체외대인효질류U87세포증식、조망화침습적영향.방법 인공합성적HIF-1α반의단발협경양리자지질체포과후순시전염인효질류세포주U87,채용Western blot법검측전염후효질류세포HIF-1α단백표체,증실전염성공,재채용새서람(MTT)비색법화Transwell소실체외침습실험검측전염후대U87세포증식체외침습능력적영향,류식세포의검측세포조망.결과 전염파향HIF-1α적단련발협RNA적효질류세포,HIF-1α단백표체교공백세포조명현강저,MTT법검측전염조、공재체조화공세포조24 h세포적증식솔분별위5.46%、21.25%、22.32%,체외침습성실험표명전염조、공재체조화공세포조세포12 h침습ECM적세포수분별위(22±4)、(124±3)、(122±6);류식세포의검측표명전염조、공재체조화공세포조세포조망솔분별위(53.35±2.80)%、(12.02±1.60)%、(10.19±3.15)%,전염조여공백세포조급공재체조비교차이균유통계학의의(P＜0.05).결론 봉폐HIF-1α단백적표체,가이억제인효질류U87세포증식화침습능력,촉진세포조망.
Objective To investigate the effects of antisense oligodeoxynucleotides (ODNs) targeting hypoxia inducible factor-1α (HIF-1α) on the apeptosis, 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 cells 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.
