Hepa1-6肝癌细胞经丝裂霉素处理后对survivin表达的影响

Expression of survivin mRNA and protein in mitomycin-treated hepatoma carcinoma Hepa1-6 cells

目的探讨经丝裂霉素(MMC)处理后Hepa1-6肝癌细胞survivin表达的变化。方法体外培养Hepa1-6经1.0、3.0和9.0μg/ml浓度的MMC处理后1和3d,MTT法测定Hepa1-6生长抑制率,RT-PCR法检测survivin mRNA和Western blot法检测survivin蛋白的表达。结果9.0μg/ml浓度的MMC对Hepa1-6的抑制率明显高于1.0和3.0μg/ml浓度组,1.0和3.0μg/ml的MMC作用3d后对Hepa1-6生长无抑制。各浓度MMC处理第1天的Hepa1-6表达survivin mRNA以及survivin均降低,1.0和3μg/ml的MMC处理第3天,Hepa1-6表达survivin mRNA以及survivin升高,9μg/ml组survivin mRNA以及survivin表达于第3天仍低。结论低浓度MMC能诱导Hepa1-6细胞内survivin表达增加,可能是Hepa1-6对化疗药物产生耐药性的因素之一。

Objective To investigate survivin mRNA and protein expressions in mitomycin （MMC）-treated hepatoma carcinoma Hepal-6 cells in vitro. Methods Hepal-6 cells were cultured in vitro in the presence of MMC at the concentrations of 1.0, 3.0 and 9.0 μg/ml, respectively, and 1 day and 3 days after the culture, the cell growth inhibition was assessed using MTT assay and the expressions of survivin were detected by RT-PCR and Western blotting. Results MMC at the concentration of 9.0 μg/ml resulted in significantly greater growth inhibition of the Hepa-6 cells than MMC at 1.0 and 3.0 μg/ml, and at the latter two concentrations, MMC treatment for 3 days did not produce obvious cell growth inhibition. Survivin expressions at both the mRNA and protein levels in Hepal-6 cells were significantly decreased 1 day after MMC treatment at the 3 concentrations, and after 3-day MMC treatment at 1.0 and 3μg/ml, survivin expressions increased to exceed the control level, whereas survivin maintained the low expression levels in cells treated with 9μg/ml MMC for 3 days. Conclusion Survivin expression in Hepal-6 cells increases in response to MMC treatment at low doses, which might be one of the reasons for chemotherapeutic drug resistance.