华蟾素注射液对HepG2细胞NF-κB通路的影响

Effect of cinobufagin on nuclear factor-κB pathway in HepG2 cells

目的探讨华蟾素注射液对肝癌细胞株HepG2NF-κB通路的影响。方法在TNF-α激活NF-κB通路的条件下利用双荧光素酶报告基因系统检测华蟾素注射液对含有NF-κB反应元件的萤火虫荧光素酶报告基因pNF-κB-TA-luc转录活性的影响。Western blotting进一步验证华蟾素注射液对HepG2细胞核内NF-κB亚基p65蛋白量的影响,同时半定量RT-PCR检测NF-κB下游靶基因细胞间黏附分子1(ICAM-1)的转录水平。结果0.25、0.5μg/ml浓度的华蟾素注射液可有效地抑制pNF-κB-TA-luc报告基因的相对荧光素酶值(P<0.05)。Western blot结果进一步证明华蟾素可使NF-κB亚基p65蛋白量降低,RT-PCR结果表明NF-κB下游靶基因ICAM-1表达降低。结论华蟾素注射液的抗癌机制可能与抑制癌细胞NF-κB通路的活化有关。

Objective To investigate the effect of cinobufagin on nuclear factor-κB （NF-κB） pathway of liver cancer cell line HepG2. Methods Dual-luciferase cis-reporting system was used to detect the relative value of pNF-κB-TA-luc upon tumor necrosis factor-α （TNF-α） stimulation of NF-κB pathway. Western blotting was used to detect the protein level of NF-κB p65, and RT-PCR was used to detect the gene transcription level of intercellular adhesion molecule-1 （ICAM-1）, a target downstream gene of NF-κB. Results At the concentration of 0.25 and 0.5 μg/ml, cinobufagin significantly lowered the relative value of luciferase （P0.05）. The results of Western blotting showed that cinobufagin significantly suppressed the protein expression of NF-κB p65. The transcription level of ICAM-1 was reduced by different doses of cinobufagin. Conclusion The anti-cancer effect of cinobufagin may be related to its activity in inhibiting the activation of NF-κB pathway.