17-羟-岩大戟内酯B对肝癌BEL-7402细胞增殖和凋亡的影响及机制  

Effect of 17-hydroxy-jolkinolide B on proliferation and apoptosis of liver cancer BEL-7402 cells and its mechanism

作　　者：龚飞 吴思明 徐磊 林宇[1] 马德翊 韩翠翠[1] Gong Fei;Wu Siming;Xu Lei;Lin Yu;Ma Deyi;Han Cuicui(School of pharmacy,Qiqihar Medical University,Qiqihar,Heilongjiang,161006,China;Department of vascular surgery,the first hospital of Qiqihar City,Qiqihar,Heilongjiang,161000,China)

机构地区：[1]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔市第一医院血管外科,黑龙江齐齐哈尔161006

出　　处：《齐齐哈尔医学院学报》2023年第8期701-705,共5页Journal of Qiqihar Medical University

基　　金：国家自然科学基金项目(81903865);齐齐哈尔市科技计划联合引导项目(LHYD-202003);黑龙江省普通本科高等学校青年创新人才培养计划项目(UNPYSCT-2018030);齐齐哈尔医学院研究生创新基金项目(QYYCX2022-36);黑龙江省大学生创新创业训练计划项目(X202211230021)。

摘　　要：目的研究17-羟-岩大戟内酯B(HJB)对人肝癌BEL-7402细胞增殖和凋亡的影响,并探讨其分子机制。方法将BEL-7402细胞分为空白对照组、HJB 10μg/mL组、20μg/mL组和40μg/mL组。采用MTT实验检测各组细胞存活率。采用流式细胞术检测HJB对BEL-7402细胞周期和凋亡的影响;AO/EB双染色法检测细胞凋亡形态的变化;Western blot法检测Bcl-2、Bax、Cleaved-Caspase-9和Cleaved-Caspase-3蛋白表达的变化。结果HJB可呈浓度-时间依赖趋势抑制BEL-7402细胞增殖。不同浓度HJB作用24小时后,各HJB处理组主要表现出S期和G2/M期阻滞;早、晚期细胞凋亡率均明显增加(P<0.05或P<0.01)。随着HJB浓度的增加,晚期凋亡细胞数量逐渐增加,呈固缩或圆珠状。各HJB处理组Bcl-2的蛋白表达均明显下调,40μg/mL组细胞Bax的蛋白表达明显上调,此外,各组细胞Cleaved-Caspase-9和Cleaved-Caspase-3蛋白的表达均显著上调(P<0.05)。结论17-羟-岩大戟内酯B可以抑制肝癌BEL-7402细胞增殖,调控细胞周期并诱导其凋亡,其分子机制可能与下调Bcl-2蛋白表达、上调Bax、Cleaved-Caspase-9和Cleaved-Caspase-3蛋白表达有关。Objective To study the effects of 17-hydroxy-jolkinolide B(HJB)on the proliferation and apoptosis of liver cancer BEL-7402 cells,and to investigate its molecular mechanism.Methods BEL-7402 cells were divided into normal group,HJB 10μg/mL group,HJB 20μg/mL group and HJB 40μg/mL group.MTT assay was used to detect the survival rate of cells.The apoptosis and cycle distribution of BEL-7402 cells were detected by flow cytometry.Apoptosis morphology was detected by AO/EB double staining.Western blot was used to detect protein expression of Bcl-2,Bax,Cleaved-Caspase-9,Cleaved-Caspase-3.Results HJB inhibited the proliferation of BEL-7402cells in a concentration-time dependent trend.After treatment for 24 h by different concentration of HJB,compared with control group,the cells of each HJB treatment group were mainly blocked in S phase and G2/M phase;the early and late apoptosis rate increased significantly(P<0.05 or P<0.01).With the increase of HJB concentration,the number of late apoptotic cells increased gradually,showing the pyknosis or spherical shape.Results of Western blot showed that the relative protein expression of Bcl-2 were reduced significantly in HJB treatment groups;the relative protein expression of Bax were increased significantly in HJB 40μg/mL group;besides,the relative protein expression of Cleaved-Caspase-9 and Cleaved-Caspase-3 were increased significantly in HJB treatment groups(P<0.05).Conclusions 17-hydroxy-jolkinolide B can significantly inhibit the proliferation,regulate cell cycle and induce apoptosis of liver cancer BEL-7402 cells.The mechanism of apoptosis may be related with down-regulating the protein expression of Bcl-2,up-regulating the protein expression of Bax,Cleaved-Caspase-9 and Cleaved-Caspase-3.

关 键 词：17-羟-岩大戟内酯B 肝癌 BEL-7402细胞 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]