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].现代生物医学进展英文版,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 |
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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) |
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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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