| 殷杰,钟苗,冯兰芳,朱炳阳,唐圣松,雷小勇.Bcl-2 siRNA增强HepG2细胞对5-氟尿嘧啶敏感性.[J].中南医学科学杂志.,2007,(4):503-506. |
| Bcl-2 siRNA增强HepG2细胞对5-氟尿嘧啶敏感性 |
| Bcl-2 siRNA Increased Sensitivity to 5-Fluorouracil in HepG2 Cells |
| 修订日期：2007-04-29 |
| DOI： |
| 中文关键词: Bcl-2 RNA干扰 HepG2细胞 5-氟尿嘧啶 |
| 英文关键词:Bcl-2,RNA interference,HepG2 cells,5-FU |
| 基金项目: |
| 殷杰 钟苗 冯兰芳 朱炳阳 唐圣松 雷小勇 |
| 南华大学药物药理研究所,湖南衡阳421001 |
| 摘要点击次数: 860 |
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| 中文摘要: |
| 目的 观察Bcl-2 siRNA对人类肝癌细胞HepG2 5-氟尿嘧啶（5-FU）敏感性的影响。方法 Bcl-2 siRNA表达载体构建并稳定转染至肝癌细胞HepG2中,用G418筛选稳定的阳性细胞克隆。用RT-PCR检测Bcl-2 mRNA水平,免疫荧光检测目的基因Bcl-2蛋白水平的表达。用131、30、1300和13 000 mg/L 5-FU处理稳定转染细胞,用MTT观察细胞对药物敏感性的影响,用流式细胞术观察细胞凋亡情况。Western blot检测Bcl-2和Bax蛋白水平。结果 5-FU处理细胞24 h,Bcl-2 siRNA转染组的细胞生长抑制率明显高于阴性载体转染组和正常细胞;Bcl-2 siRNA转染组细胞凋亡率高于正常细胞和阴性载体转染组;在Bcl-2表达下调时,Bax蛋白表达水平不变。结论 siRNA下调目的基因Bcl-2能增强人类肝癌细胞HepG2对5-FU的敏感性;HepG2细胞对5-FU的敏感性增强与Bax/Bcl-2的比例增高有关。 |
| 英文摘要: |
| Objective To investigate the changes of sensitivity to 5-fluorouracil (5-FU)in Bcl-2 siRNA transfected Hepg2 cells.Method Bcl-2 siRNA expression vector were constructed and stably transfected into HepG2 cells.RT-PCR and Immunofluorescence were used to detect the target gene expression.Western Blotting was used to detect Bcl-2,Bax expressiom.Drug sensitivity of the cells to 5-fluorouracil were analyzed with MTT and flow cytometry.Results MTT results showed that Bcl-2 transfectants had a higher cell inhibition rate than negative vector or untreated cells after treated with13,130,1 300,13 000 mg/L 5-FU;Moreover,Flow cytometry results demonstrated that Bcl-2 siRNA cells had increased apoptosis rate compared with negative siRNA or untreated cell;The protein expression of Bax had no change when Bcl-2 protein was reduced.Conclusion siRNA targeting Bcl-2 gene can specifically down-regulate Bcl-2 expression,increased Bax/Bcl-2 ratio expression which lead to increase cell spontaneous apoptosis and sensitize cells to 5-FU. |
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