文章摘要
王希超 刘相萍 温艳玲 吕志栋 薛丹丹 王海波.抑制c-FLIP基因促进TRAIL诱导的乳腺癌细胞凋亡(英文)[J].,2014,14(23):4425-4429
抑制c-FLIP基因促进TRAIL诱导的乳腺癌细胞凋亡(英文)
Down-regulation of c-FLIP Gene by siRNA Interference Enhances Breast Cancer Cells toTRAIL-Induced Apoptosis
  
DOI:
中文关键词: TRAIL  c-FLIP  乳腺癌  腺病毒  凋亡
英文关键词: TRAIL  c-FLIP  Breast cancer  Adenovirus  Apoptosis
基金项目:山东省自然科学基金项目(2008C48);山东省教育厅资助项目(J11LF05);国家自然科学基金项目(81302290)
作者单位
王希超 刘相萍 温艳玲 吕志栋 薛丹丹 王海波 青岛大学医学院附属医院乳腺诊疗中心青岛大学医学院附属医院中心实验室 青岛大学医学院附属医院耳鼻喉科 
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中文摘要:
      目的:探讨抑制c-FLIP 的表达对TRAIL 诱导乳腺癌细胞MCF-7 凋亡的影响。方法:重组腺病毒Ad-c-FLIP-siRNA 和 Ad-sTRAIL单独及联合感染对TRAIL 耐药的乳腺癌细胞MCF-7,应用实时荧光定量聚合酶链反应(Real-time PCR)检测病毒感 染后各组细胞内c-FLIP 和TRAIL 的mRNA表达变化;MTT 法和结晶紫染色法检测MCF-7 细胞活性,Hoechst 33258 荧光染色 检测各组细胞的凋亡情况。结果:与阴性对照组比较,c-FLIP-siRNA组和c-FLIP-siRNA+TRAIL组c-FLIP 的mRNA相对表达量分 别是(0.32± 0.16)和(0.39± 0.48)倍;TRAIL 组和c-FLIP-siRNA+TRAIL 组TRAIL 的mRNA 相对表达量分别是(96.21± 1.54)和 (87.33± 1.66)倍;TRAIL 组、c-FLIP-siRNA 组及c-FLIP-siRNA+TRAIL 组的抑制率(%)分别为(60.27± 1.25)、(11.34± 1.74)及 (74.91± 2.12)。对比阴性对照组的凋亡率(3.12± 1.54),TRAIL 组(12.79± 2.46)和c-FLIP-siRNA+TRAIL 组(25.50± 3.17)组的凋 亡率明显增高(P<0.05),c-FLIP-siRNA 组(6.85± 2.82)的凋亡率变化不明显,差异无统计学意义(P>0.05)。结论:siRNA 抑制 c-FLIP 基因的表达能显著促进TRAIL对乳腺癌细胞MCF-7 凋亡的诱导作用。
英文摘要:
      Objective:To investigate the effect of c-Flip gene suppression on enhancing breast cancer cell MCF-7 to TRAIL-induced apoptosis.Methods:MCF-7 cells were infected with recombinant adenoviruses Ad-cFLIP-siRNA and Ad-sTRAIL, alone or combined respectively. Real-time quantitative polymerase chain reation (Real time PCR) was used to detect the expression of c-FLIP and TRAIL in MCF-7 cells. Methythiazol tetrazolium (MTT) assay and Crystal violet staining were applied to detect the cell proliferation. And Hoechst 33258 staining was used to detect the apoptosis of MCF-7 cells.Results:Compared with that in the negative control group, the relative expressions of c-FLIP in the c-FLIP-siRNA and the c-FLIP-siRNA+TRAIL groups were (0.32± 0.16) and (0.39± 0.48) times respectively; The relative expressions of TRAIL in the TRAIL and the c-FLIP-siRNA+TRAIL groups were (96.21± 1.54) and (87.33± 1.66) times respectively. The inhibition rate (%) of cells in TRAIL, c-FLIP-siRNA, and TRAIL+c-FLIP-siRNA group were (60.27± 1.25), (11.34± 1.74) and (74.91± 2.12), respectively. Compared with that in the negative control group(3.12± 1.54), the apoptosis rates of cells in TRAIL (12.79± 2.46) and c-FLIP-siRNA+TRAIL group (25.50± 3.17) increased significantly (P<0.05). However, the apoptosis rates of cells in c-FLIP-siRNA group (6.85± 2.82) had no statistical significance with that in negative control group (P>0.05).Conclusion:Down-regulation of c-FLIP by siRNA interference could enhance the breast cancer cells MCF-7 to TRAIL-induced apoptosis.
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