| 王胜陈云芳付欣洪伟李冰.β- 萘黄酮能抑制荧光素酶活性[J].,2011,11(20):3853-3856 |
| β- 萘黄酮能抑制荧光素酶活性 |
| β-Naphthoflavone Can Inhibit Luciferase Activity |
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| DOI： |
| 中文关键词: β- 萘黄酮 基因转录调控 荧光素酶 报告基因 |
| 英文关键词: β-Naphthoflavone Gene transcriptional regulation Luciferase Reporter gene |
| 基金项目:国家自然科学基金项目资助（30971166） |
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| 中文摘要: |
| 目的：研究β- 萘黄酮对荧光素酶活性的影响。方法：利用人GCLC 基因调控序列驱动的GCLC-PGL3-enhancer-Luciferase 报
道载体（PL45）转染人肺腺癌细胞A549，人肝癌细胞HepG2，人子宫颈癌细胞HeLa，人乳腺癌细胞MCF-7，人肝癌细胞Bel-7402，
人支气管上皮细胞16HBE，β- 萘黄酮刺激后，双荧光素酶报告基因检测系统分析其对GCLC 基因表达的影响。wester blot 检测
β-NF 刺激16HBE 细胞后GCLC 蛋白水平的变化。β- 萘黄酮刺激转染了表达Luciferase 的真核表达载体pRC/CMV2- luc+ 的
A549 和HepG2 细胞后，双荧光素酶报告基因检测系统分析其对Luciferase 基因表达的影响。PL45 转染A549 和HepG2 细胞，裂
解细胞后用β-NF 刺激，双荧光素酶报告基因检测系统分析其对Luciferase 基因的影响。结果：在各种细胞中，转染PL45 报道载
体后，β-NF 处理组荧光素酶相对活性值与DMSO 对照组相比均明显下降（P<0.01）。wester blot 结果显示β-NF 处理组GCLC 蛋
白的表达较DMSO 对照组明显升高。在A549 和HepG2 细胞中，转染pRC/CMV2- luc+ 载体后，β-NF 处理组荧光素酶相对活性
值与DMSO 对照组相比均明显下降（P<0.01）。PL45 转染A549 和HepG2 细胞，裂解细胞后用β-NF 刺激，β-NF 处理组荧光素酶
相对活性值与DMSO 对照组相比均明显下降（P<0.01）。结论：β- 萘黄酮直接抑制了荧光素酶的活性 |
| 英文摘要: |
| Objective: To investigate the effect of β-Naphthoflavone on the firefly luciferase activity. Methods: A549, HepG2,
HeLa, MCF-7, Bel-7402, 16HBE cells were transfected with GCLC5'-upstream regulatory sequence driven PGL3-enhancer-Luciferase
reporter vector （PL45）and treated with β-NF. The dual-luciferase reporter assay system was used to analyze the effect of
β-Naphthoflavone on the expression of GCLC gene. Western blot was used to detect the change of protein level. A549 and HepG2 cells
were transfected with the eukaryotic expression vector pRC/CMV2- luc+ and treated with β-NF. The dual-luciferase reporter assay
system was used to analyze the effect of β-Naphthoflavone on the firefly luciferase activity. A549 and HepG2 cells were transfected with
the PL45 vector, cells were lysed, then treated with β-NF for 25 min to analyze the effect of β-NF on the firefly luciferase activity.
Results: In all the cells, the relative luciferase activity of β-NF treatment group was significantly lower compared with that in DMSO
control group （P<0.01）. The result of western bot showed that the expression of GCLC were higher inβ-NF treatment group than that in
DMSO control group. In A549 and HepG2 cells, after transfected with pRC/CMV2- luc+ vector, the relative luciferase activity of β-NF
treatment group was lower than that in DMSO control group （P<0.01）.A549 and HepG2 cells were transfected with PL45 vector, then
lysed, and the relative luciferase activity in β-NF treatment group was lower than that in DMSO control group（P<0.01）.Conclusion:
β-Naphthoflavone directly inhibits Firefly Luciferase activity. |
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