橙皮素衍生物对肝癌细胞SMMC-7721的促凋亡作用

发布时间：2018-02-27 05:23

本文关键词： 肝癌人肝癌细胞SMMC- 橙皮素 Bcl- Bax　出处：《安徽医科大学学报》2017年07期 　论文类型：期刊论文

【摘要】：目的以橙皮素为原型,合成了新型化合物HY-12,研究其体外对SMMC-7721细胞的促凋亡作用及机制。方法在3个时间点(24、48、72 h)采用MTT比色法观察不同浓度HY-12对肝癌细胞、肝细胞增殖的影响,计算半数抑制率(IC50)。根据结果选取了24 h,12.5×10-6,25×10-6,50×10-6mol/L 3个浓度进行下述实验。通过Annexin V/PI双染,流式细胞仪检测细胞凋亡率;制备细胞爬片,Hoechst 33342染色,在荧光倒置显微镜下观察细胞凋亡的形态学改变;采用Western blot法检测细胞凋亡相关因子Bcl-2、Bax蛋白水平的表达情况。结果 (1)MTT法显示在对肝细胞活性无显著影响的前提下,HY-12在体外可抑制SMMC-7721细胞的增殖,且具有剂量-时间依赖性;(2)流式细胞仪检测显示HY-12作用后,SMMC-7721细胞凋亡率增加,且呈浓度依赖性趋势;(3)经Hoechst 33342染色,药物处理后细胞核皱缩断裂呈现亮蓝色新月状;(4)Western blot结果显示药物处理可以使Bax蛋白表达上调,相反地Bcl-2蛋白表达下调。结论橙皮素衍生物HY-12有明显抑制肝癌细胞SMMC-7721增殖作用。其机制可能是通过调节Bcl-2、Bax表达以诱导细胞凋亡。HY-12可能成为治疗肝癌的有效化疗药物。
[Abstract]:Objective to synthesize a novel compound HY-12 from hesperidin, and to study its effect on apoptosis of SMMC-7721 cells in vitro and its mechanism. Methods MTT colorimetric assay was used to observe the effects of different concentrations of HY-12 on the proliferation of hepatoma cells and hepatocytes. The 50% inhibition rate (IC50) was calculated. According to the results, the following three concentrations were selected: 24 h, 12.5 脳 10 -6, 25 脳 10 -6, 50 脳 10 -6 mol / L, respectively. The apoptosis rate was detected by Annexin V / Pi double staining, and the cell climbing slices were prepared for Hoechst 33342 staining. The morphological changes of apoptosis were observed under fluorescence inverted microscope. The expression of apoptosis-related factor Bcl-2 Bax protein was detected by Western blot assay. Results the results showed that HY-12 could inhibit the proliferation of SMMC-7721 cells without significantly affecting the hepatocyte activity in vitro. Flow cytometry analysis showed that the apoptosis rate of SMMC-7721 cells increased after HY-12 treatment, and showed a concentration-dependent trend. Hoechst 33342 staining showed that the apoptosis rate of SMMC-7721 cells was higher than that of SMMC-7721 cells in a dose-dependent manner. The results of Western blot showed that the nuclear shrinkage and cleavage after drug treatment could up-regulate the expression of Bax protein. On the contrary, the expression of Bcl-2 protein was down-regulated. Conclusion HY-12, a derivative of hesperidin, can significantly inhibit the proliferation of SMMC-7721 in hepatocellular carcinoma cells, and its mechanism may be that it may be an effective chemotherapeutic drug for the treatment of hepatocellular carcinoma by regulating the expression of Bcl-2P Bax to induce apoptosis.
【作者单位】：安徽医科大学药学院安徽医科大学肝病研究所安徽省创新药物产业共性研究院;
【基金】：安徽省高等学校省级自然科学研究重点项目(编号:KJ2016A364、KJ2016A365) 安徽省自然科学基金面上项目(编号:1408085MKL31) 国家自然科学基金(编号:81273526、81473268)
【分类号】：R96

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