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目的:研究莱菔硫烷(Sulforaphane,SFN)在体外对HepG-2细胞Bcl-2、Bax基因转录和蛋白表达的影响,探讨SFN诱导人肝癌HepG-2细胞凋亡的机制。方法:不同浓度SFN处理体外培养的HepG-2细胞株48 h,采用SRB法测定SFN对HepG-2细胞增殖的影响;激光共聚焦显微镜观察凋亡细胞形态;流式细胞仪检测SFN作用后的细胞凋亡率;Western blot法和RT-PCR法检测SFN在蛋白和mRNA水平上对Bcl-2和Bax表达的影响。结果:SFN对HepG-2细胞的GI50为9.40μmol/L,TGI为26.68μmol/L;10、20、40μmol/L的SFN作用于HepG-2细胞48 h后,激光共聚焦显微镜下呈现典型的凋亡细胞形态,细胞凋亡率分别为(27.42±0.43)%、(46.53±0.35)%和(58.92±0.48)%(P<0.01);细胞内Bcl-2蛋白和mRNA表达水平均显著降低(P<0.01),而20、40μmol/L的SFN作用后细胞内Bax蛋白和mRNA的表达水平均显著升高(P<0.01),Bcl-2/Bax和Bcl-2 mRNA/Bax mRNA均随SFN剂量的增加显著降低(P<0.01),且变化趋势均呈一定的剂量依赖关系。结论:SFN能抑制HepG-2细胞的增殖,诱导HepG-2细胞凋亡,可在翻译和转录水平下调Bcl-2、上调Bax的表达,这可能是SFN诱导HepG-2细胞凋亡的主要机制之一。
AIM: To investigate the effects of sulforaphane (SFN) on the transcription and protein expression of Bcl-2 and Bax in HepG-2 cells in vitro and to explore the mechanism of SFN-induced apoptosis in HepG-2 cells. Methods: HepG-2 cells were treated with different concentrations of SFN for 48 h. The effect of SFN on the proliferation of HepG-2 cells was detected by SRB method. The morphology of apoptotic cells was observed by laser confocal microscopy. The effect of SFN was detected by flow cytometry The apoptotic rate was detected by Western blot and RT-PCR. The effect of SFN on the expression of Bcl-2 and Bax protein and mRNA was examined. Results: The GI50 of SFN on HepG-2 cells was 9.40 μmol / L and the TGI was 26.68 μmol / L. After treated with 10, 20 and 40 μmol / L SFN for 48 h, the typical expression of SFN was observed under confocal laser scanning microscope (27.42 ± 0.43)%, (46.53 ± 0.35)% and (58.92 ± 0.48)%, respectively (P <0.01). The expression of Bcl-2 protein and mRNA in the cells were significantly decreased (P <0.01). However, the expression of Bax protein and mRNA in the cells treated with 20,40 μmol / L SFN increased significantly (P <0.01), and the mRNA and protein expressions of Bcl-2 / Bax and Bcl- The increase of SFN dose was significantly lower (P <0.01), and the change trend showed a dose-dependent manner. Conclusion: SFN can inhibit the proliferation of HepG-2 cells, induce the apoptosis of HepG-2 cells, down-regulate Bcl-2 and up-regulate the expression of Bax at the level of translation and transcription, which may be the main mechanism of SFN-induced HepG-2 cell apoptosis one.