| 摘要: |
| 【目的】探讨红景天甙(Salidroside)体外诱导人肝癌QGY-7703细胞凋亡的作用及其可能的机制。【方法】-以体外培养的人肝癌QGY-7703细胞作为研究对象，采用四甲基偶氮唑盐（MTT）比色法检测红景天甙对QGY-7703细胞生长的抑制作用，采用吖啶橙/溴化乙啶(AO/EB)荧光双染、琼脂糖凝胶电泳及流式细胞术，分析红景天甙对QGY-7703细胞凋亡的诱导作用；采用Western blot免疫印迹法探讨红景天甙诱导QGY-7703细胞凋亡的可能机制。【结果】-MTT比色法试验结果表明，0.01，0.1，1，10和100 μg/mL红景天甙对QGY-7703细胞生长均有不同程度的抑制作用，细胞生长抑制率分别为13.2%，21.9%，31.4%，46.8%和60.9%，半数抑制质量浓度(IC50)为18.56 μg/mL；用10 μg/mL 红景天甙处理QGY-7703细胞48 h，AO/EB荧光双染后，可观察到个别QGY 7703细胞呈现亮绿色或橘红色，表明细胞发生了凋亡；琼脂糖凝胶电泳结果显示，10 μg/mL红景天甙作用QGY-7703细胞24，48 和72 h均可导致细胞核内DNA产生有序断裂，形成DNA梯形带；流式细胞术试验结果表明，10 μg/mL红景天甙的加入量为100 和200 μL时，QGY-7703细胞凋亡数分别为178和331个，相应的凋亡率分别为1.8%和3.5%，阴性对照组（10 μg/mL红景天甙加入量为0 μL）凋亡细胞数为105个，凋亡率仅为1.1％；Western blot免疫印迹分析结果表明，红景天甙下调了Bcl-2和PCNA蛋白的表达量，而使凋亡活化基因Bax与肿瘤抑制基因p53的蛋白表达量增高。【结论】 红景天甙对体外培养的QGY-7703细胞具有明显的生长抑制作用和诱导凋亡作用，其诱导凋亡作用与Bcl-2家族成员、p53、Bax和PCNA基因调控有关。 |
| 关键词: 红景天甙 体外诱导 QGY 7703细胞 细胞凋亡 |
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| 基金项目:西北民族大学校中青年科研基金项目(XBMU-2006-BD-84) |
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| Study on cell apoptosis of human hepatoma QGY-7703 cell induced by salidroside in vitro culture |
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| Abstract: |
| 【Objective】The experiment was conducted to study cell apoptosis and possible mechanism of human hepatoma QGY-7703 cells induced by salidroside in vitro culture.【Method】With human hepatoma QGY-7703 cells as study object in vitro culture,the QGY 7703 growth inhibition effect of salidroside was measured by methyl thiazolyl tetrazolium (MTT) method;The apoptosis effect of QGY-7703 was analyzed by AO/EB double fluorescent dye staining,agarose gel electrophoresis and flow cells analysis;The possible mechanism of QGY-7703 apoptosis induced by salidroside was studied by western blot.【Result】The result of MTT method showed that salidroside of 0.01,0.1,1,10 and 100 μg/mL all inhibited the growth of QGY-7703 cells.The cell growth inhibition rate of QGY-7703 was 13.2%,21.9%,31.4%,46.8% and 60.9% respectively and the 50% inhibiting concentration (IC50) was 18.56 μg/mL.By AO/EB double fluorescent dye staining treated with 10 μg/mL salidroside for 48 h,individual cells presented bright green and jacinth.It showed that these cells apoptosised.The DNA in nucleus appeared regular rupture and formed DNA ladder treated with 10 μg/mL salidroside for 24,48,and 72 h in agarose gel electrophoresis experiment.The result of flow cells analysis showed that the apoptosis cells were 178 and 331 and the apoptosis rate was 1.8% and 3.5% respectively when added amount of 10 μg/mL salidroside was 100 μL and 200 μL,but the apoptosis cells were 105 and the apoptosis rate was only 1.1% in the control group(the volume of 10 μg/mL salidroside was 0 μL).Western blot showed that salidroside down-regulated the expression of Bcl-2 and PCNA proteins,but protein expressions of apoptosis activation gene Bax and tumor suppressor gene p53 were up-regulated.【Conclusion】The salidroside could inhibit cell growth and induce cell apoptosis of OGY-7703 cells in vitro culture.The effect of apoptosis was related to the gene modulation of Bcl-2 family,p53,Bax and PCNA. |
| Key words: Salidroside induction in vitro QGY-7703 cell cell apoptosis |
