β-榄香烯诱导人结肠癌细胞DLD-1的增殖抑制和凋亡研究  

Study of β-elemene on tumor inhibition and apoptosis of human colon cancer DLD-1 cells

作　　者：汪国玉 张蕾[2] 耿亚迪 冯晓俊[2] 陈昭琳 赵晓晓[1] 姜玲[1,2] 魏伟[1] Wang Guoyu;Zhang Lei;Geng Yadi(Dept of Clinical Pharmacology;o f Anhui Medical University, Key Laboratory o f Anti-inflammatory and ImmuneMedicine o f Education Ministry, Anhui Collaboratiw Innovation Center o f Anti-inflammatory and Immune M edicine,Hefei 230032;2Dept Pharm acy, The Affiliated Provincial Hospital 0^ Anhui MePical University, Hefei 230001)

机构地区：[1]安徽医科大学临床药理研究所抗炎免疫药物教育部重点实验室抗炎免疫药物安徽省协同创新中心,合肥230032 [2]安徽医科大学附属安徽省立医院药剂科,合肥230001

出　　处：《安徽医科大学学报》2018年第4期491-497,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81603339;81602344);安徽省自然科学基金(编号:1708085QH175)

摘　　要：目的研究β-榄香烯对人结肠癌细胞DLD-1的增殖、凋亡及对活性氧(ROS)的影响。方法体外培养人结肠癌细胞DLD-1,采用MTS法、克隆形成实验检测不同浓度β-榄香烯对DLD-1细胞增殖的影响,使用Hoechst 33342染色、流式细胞术检测β-榄香烯对DLD-1细胞凋亡的影响,Western blot法检测凋亡相关蛋白表达水平的变化,DCFH-DA荧光染色检测ROS水平。结果 MTS法和克隆形成实验显示β-榄香烯对DLD-1细胞具有增殖抑制作用;Hoechst 33342染色显示β-榄香烯可使细胞出现细胞核聚缩和致密浓染的表现,Annexin V/PI双染结果显示,β-榄香烯可诱导DLD-1细胞出现凋亡;Western blot结果显示β-榄香烯作用后Cleaved PARP、Cleaved Caspase-9和Cleaved Caspase-3表达明显上调;ROS的测定结果显示β-榄香烯可使细胞内ROS水平显著升高。结论β-榄香烯能显著抑制人结肠癌细胞DLD-1增殖并能够促进细胞凋亡,ROS升高可能是其发挥作用的机制之一。Objective To investigate effects of β-elemene on the proliferation and apoptosis of human colon cancer DLD-1 cell line and its effect on reactive oxygen species（ ROS）. Methods Human colon cancer DLD-1 cell line was cultured in vitro. MTS and cell clone forming experiments were performed to reveal the inhibitory effect on DLD-1 cells proliferation with different concentrations of β-elemene. Hoechst 33342 staining and flow cytometry were used to determine the apoptosis of DLD-1 cells after treated with β-elemene,and the protein expression levels of polymerase（ PARP）,the cleavage of PARP（ Cleaved PARP）,Cleaved Caspase-9,Caspase-9,Cleaved Caspase-3 and Caspase-3 were determined by Western blot. DCFH-DA fluorescence staining was used to detect the level of ROS. Results MTS and cell clone forming experiments showed that β-elemene significantly inhibited the proliferation of DLD-1 cells. Hoechst 33342 staining showed that β-elemene caused DLD-1 cells nucleus condensation and dense staining. Annexin V/PI double staining showed that β-elemene could induce DLD-1 cells apoptosis.Western blot showed that the protein expression levels of Cleaved PARP,Cleaved Caspase-9 and Cleaved Caspase-3 were significantly up-regulated after β-elemene treatment. ROS assay showed that β-elemene obviously elevated ROS level of DLD-1 cells. Conclusion β-elemene significantly inhibited DLD-1 cells proliferation and promoted DLD-1 cells apoptosis,and the increasement of ROS level may play one of the mechanisms.

关 键 词：Β-榄香烯 人结肠癌细胞 增殖 凋亡 

分 类 号：R965.2[医药卫生—药理学]