摘要
为探讨异甘草素对小鼠黑色素瘤B16F0细胞诱导凋亡的影响。采用SRB法检测异甘草素对B16F0细胞增殖的影响;台盼蓝拒染法检测细胞致死率;吖啶橙/溴乙啶荧光染色法观察细胞凋亡形态;AO/EB和Hoechst 33258染色观察药物处理后细胞形态的变化;流式细胞仪Annexin V-FITC/PI双染发检测细胞凋亡率;caspase-9/3试剂盒检测半胱氨酸蛋白酶-9和半胱氨酸蛋白酶-3的活性;QPCR法检测细胞凋亡相关基因B淋巴细胞-2,Bcl-2相关X蛋白的m RNA表达。结果显示,异甘草素能有效抑制B16F0细胞增殖,呈现浓度依赖性和时间依赖性,作用于B16F0细胞后呈现出明显的凋亡形态,同时随着异甘草素浓度的增加(0、24、45、65μg/m L),细胞凋亡率增加;caspase-9,caspase-3活性逐渐升高;Bax/Bcl-2比率上调(P<0.05或P<0.01)。由此可知,异甘草素能够显著诱导小鼠黑色素瘤B16F0细胞凋亡,抑制肿瘤细胞增殖。
To investigate the effects of isoliquiritigenin（ISL） on apoptosis in mouse melanoma cell line B16F0. Sulforhodamine B（SRB） was used to test cell viability. Trypan blue exclusion method was utilized into determining cell fatality rates of B16F0 cells. Acridine orange/ethidium bromide（AO/EB） and Hoechst 33258 staining were employed to observe cell morphological changes. The cell apoptotic rate was determined by staining with annexin V flourescein isothiocyanate（Annexin V-FITC） and propidium iodide（PI） in flow cytometry. Activity changes of caspase-9 and caspase-3 were measured by commercial kit, and the expression rate of Bax/Bcl-2 was detected using Real-Time quantitative PCR. The proliferation of B16F0 cells treated with ISL was effectively inhibited in a concentration and time-dependent manner. The result of fluorescence microscopic imaging demonstrated that the typical apoptotic morphology was observed in the ISL-treated group. With the increasing concentration of ISL, the number of apoptotic cells increased as well. Activities of caspase-9 and caspase-3 enhanced in the ISL-treated B16F0 cells. The expression rate of Bax/Bcl-2 was pushed up by ISL. These findings in our research show that ISL significantly induces apoptosis of mouse melanoma B16F0 cells and thus inhibits their proliferation.
出处
《石河子大学学报（自然科学版）》
CAS
2016年第2期194-200,共7页
Journal of Shihezi University(Natural Science)
基金
国家自然科学基金项目(31471338)
新疆兵团创新研究团队基金项目(2015BD005)
滨州医学院高层次人才创新基金项目(BY2014KYQD01)
关键词
B16F0细胞
异甘草素
细胞凋亡
Mouse melanoma B16F0 cells
isoliquiritigenin（ISL）
apoptosis
