中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2011年
7期
649-653
,共5页
神经胶质瘤%低氧诱导因子1α%U87细胞%小发夹RNA
神經膠質瘤%低氧誘導因子1α%U87細胞%小髮夾RNA
신경효질류%저양유도인자1α%U87세포%소발협RNA
Glioma%Hypoxia-inducible factor-1α%U87%ShRNA
目的 观察沉默低氧诱导因子1α(HIF-1a)基因后对胶质母细胞瘤U87细胞增殖、侵袭及转移能力的影响.方法 实验分3组,干扰组(转染表达HIF-1α-shRNA的质粒)、对照干扰组(转染阴性对照shRNA序列)及未处理组.干扰组利用前期构建的HIF-1α基因的短发夹RNA(shRNA)沉默HIF-1α基因,构建HIF-1α-shRNA慢病毒表达载体,在脂质体介导下转染人胶质母细胞瘤细胞U87.采用RT-PCR和Western blotting检测HIF-1α基因干扰效率,MTT法检测细胞生长增殖能力,迁移实验检测细胞的体外迁移能力,Transwell小室模型检测细胞的体外侵袭及转移能力.结果 RT-PCR和Western blotting实验证实,与对照干扰组及未处理组比较,干扰组细胞HIF-1αmRNA表达水平明显下降,蛋白条带明显减弱.MTT细胞增殖实验显示,干扰组细胞增殖水平较其他2组明显降低,差异有统计学意义(P<0.05).Transwell小室侵袭实验中,未处理组、对照干扰组、干扰组穿膜细胞数分别为(125.2±10.8)个、(118.3±8.3)个、(60.9±5.4)个,差异有统计学意义(P<0.05).结论 HIF-1α-shRNA能有效抑制U87细胞株HIF-1α mRNA及蛋白的表达,并能抑制U87细胞的增殖、侵袭及转移能力.
目的 觀察沉默低氧誘導因子1α(HIF-1a)基因後對膠質母細胞瘤U87細胞增殖、侵襲及轉移能力的影響.方法 實驗分3組,榦擾組(轉染錶達HIF-1α-shRNA的質粒)、對照榦擾組(轉染陰性對照shRNA序列)及未處理組.榦擾組利用前期構建的HIF-1α基因的短髮夾RNA(shRNA)沉默HIF-1α基因,構建HIF-1α-shRNA慢病毒錶達載體,在脂質體介導下轉染人膠質母細胞瘤細胞U87.採用RT-PCR和Western blotting檢測HIF-1α基因榦擾效率,MTT法檢測細胞生長增殖能力,遷移實驗檢測細胞的體外遷移能力,Transwell小室模型檢測細胞的體外侵襲及轉移能力.結果 RT-PCR和Western blotting實驗證實,與對照榦擾組及未處理組比較,榦擾組細胞HIF-1αmRNA錶達水平明顯下降,蛋白條帶明顯減弱.MTT細胞增殖實驗顯示,榦擾組細胞增殖水平較其他2組明顯降低,差異有統計學意義(P<0.05).Transwell小室侵襲實驗中,未處理組、對照榦擾組、榦擾組穿膜細胞數分彆為(125.2±10.8)箇、(118.3±8.3)箇、(60.9±5.4)箇,差異有統計學意義(P<0.05).結論 HIF-1α-shRNA能有效抑製U87細胞株HIF-1α mRNA及蛋白的錶達,併能抑製U87細胞的增殖、侵襲及轉移能力.
목적 관찰침묵저양유도인자1α(HIF-1a)기인후대효질모세포류U87세포증식、침습급전이능력적영향.방법 실험분3조,간우조(전염표체HIF-1α-shRNA적질립)、대조간우조(전염음성대조shRNA서렬)급미처리조.간우조이용전기구건적HIF-1α기인적단발협RNA(shRNA)침묵HIF-1α기인,구건HIF-1α-shRNA만병독표체재체,재지질체개도하전염인효질모세포류세포U87.채용RT-PCR화Western blotting검측HIF-1α기인간우효솔,MTT법검측세포생장증식능력,천이실험검측세포적체외천이능력,Transwell소실모형검측세포적체외침습급전이능력.결과 RT-PCR화Western blotting실험증실,여대조간우조급미처리조비교,간우조세포HIF-1αmRNA표체수평명현하강,단백조대명현감약.MTT세포증식실험현시,간우조세포증식수평교기타2조명현강저,차이유통계학의의(P<0.05).Transwell소실침습실험중,미처리조、대조간우조、간우조천막세포수분별위(125.2±10.8)개、(118.3±8.3)개、(60.9±5.4)개,차이유통계학의의(P<0.05).결론 HIF-1α-shRNA능유효억제U87세포주HIF-1α mRNA급단백적표체,병능억제U87세포적증식、침습급전이능력.
Objective To observe the influence of silencing hypoxia-inducible factor-1α(HIF-1α)gene on the proliferation, invasion and metastasis of glioblastoma U87 cells. Methods The samples were divided into 3 groups: blank group: samples without giving any treatments, control group: cells with empty shRNA vector, and experimental group: cells with HIF-1α-shRNA transfection complex. HIF-1α gene was silenced by shRNA constructed in early time; and HIF-1α-shRNA lentivirus vector was constructed in the experimental group, and then transfected into glioblastoma U87 cells with the mediation of liposome. The interference efficiency was detected by using RT-PCR and Western blotting, and cell proliferation was measured by MTT assay; cell migration in vitro was observed by migration test, and invasion and metastasis abilities were detected by Transwell booth model. Results As compared with those in cells of the control and blank groups, the mRNA and protein expressions of HIF-1α in cells of the experimental group were significantly decreased; MTT assay showed that the cell proliferation in the experimental group was significantly lower than that in the other 2 groups (P<0.05). The number of penetrating cells of the blank group, control group and experimental group in Transwell chamber invasion assay were (125.2±10.8), (118.3±8.3), (60.9±5.4), respectively, and significant differences were noted between each 2 groups (P<0.05). Conclusion The mRNA and protein levels of HIF-1α in U87 cells are efficiently depressed by HIF-1α-shRNA, and so are the proliferation, invasion and metastasis abilities of U87 cells.
