贵州医科大学学报

2019, v.44;No.229(10) 1140-1144

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二氢杨梅素对肺癌A549细胞增殖和凋亡的影响
Effect of Dihydromyricetin on Proliferation and Apoptosis of Lung Cancer A549 Cells

刘金河;葛章文;张望明;晏文涛;刘志;吴昌学;
LIU Jinhe;GE Zhangwen;ZHANG Wangming;YAN Wentao;LIU Zhi;WU Changxue;National & Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Center of Tissue Engineering and Stem Cell Research, Guizhou Provincial Key Laboratory for Regenerative Medicine, Guizhou Medicine University;Key Laboratory of Adult Stem Cell Translational Research, Chinese Academy of Medical Sciences;Department of Laboratory Medicine, Guizhou Provincial People's Hospital;The First Affiliated Hospita

摘要(Abstract):

目的:探讨二氢杨梅素(DMY)对肺癌A549细胞增殖、凋亡的影响。方法:将肺癌A549细胞分为DMY组和对照组,DMY组分成5组,分别加入终浓度1、5、10、15及20 mg/L的DMY,对照组加入等量DMSO;各组细胞培养72 h时,采用MTT法检测细胞增殖抑制率,计算DMY对A549细胞的半数致死量(IC_(50)值);选取IC_(50)值附近浓度的DMY处理细胞,分别在24、48及72 h时,采用MTT检测细胞存活率;肺癌A549细胞分为DMY组(终浓度大约为IC_(50)值的DMY)及对照组,细胞培养48 h时,采用Hoechst 33342/PI双染法结合倒置荧光显微镜检测细胞凋亡,DNA ladder凝胶电泳检测细胞内DNA的断裂程度。结果:DMY能显著抑制A549细胞的增殖,细胞抑制率随着DMY浓度的增加逐渐增高(P<0.01),呈明显的浓度依赖性,IC_(50)值为(13.16±0.098)mg/L;以15 mg/L DMY处理A549细胞,细胞存活率随着时间的延长逐渐降低(P<0.01),呈明显的时间依赖性;15 mg/L DMY孵育A549细胞48 h,Hoechst 33342/PI染色后荧光显微镜下细胞核内呈现强蓝光,说明细胞处于凋亡状态,DNA ladder凝胶实验检测发现凋亡DNA片段明显增加,出现Ladder现象。结论:DMY能抑制肺癌A549细胞增殖并促进其凋亡,可能与染色体DNA断裂有关。
Objective: To observe the effect of dihydromyricetin(DMY) on proliferation and apoptosis of lung cancer A549 cells, and to explore its apoptostic mechanism. Methods: Lung cancer A549 cells cultured in vitro to logarithmic growth stage were divided into the control group and the experimental group. The latter was divided into 5 subgroups, and the final concentration of 1, 5, 10, 15, 20 mg/L was added to the five groups respectively, and the blank control group was added to the equal concentration of DMSO. Cell proliferation inhibition rate was measured by MTT assay after the culture for 72 h, and 50% lethal dose of DMY to A549 cells(IC_(50) value) was calculated. Cells treated with DMY near the IC50 value were selected, and the cell survival rate was detected by MTT 24 h, 48 h and 72 h later respectively. Lung cancer A549 cells were divided into two groups: the drug treatment group was treated with DMY near the IC50 value, and the control group was treated with DMSO with the same concentration as the drug treatment group, and 48 h after the culture of cells, Hoechst 33342/PI double staining method combined with inverted fluorescence microscope was used to detect cell apoptosis. DNA ladder gel electrophoresis was used to detect the degree of DNA breakage in apoptotic cells. Results: DMY significantly inhibited the proliferation of A549 cells in a concentration-dependent manner(P<0.01), and the IC_(50) value was(13.16±0.098) mg/L. The proliferation of A549 cells in the 24(th) h, 48(th) h, 48(th) h and 72(th) h and 72(th) h was detected and the inhibition rate was calculated in the final concentration of 15 mg/L. After staining with Hoechst33342/PI, A549 cells were incubated with DMY for 48 h and strong blue light was produced in the nucleus, indicating that the cells were in a state of apoptosis. After 48 h incubation with DMY and A549 cells, ladder gel test showed that there were only one strip in the control group, while apoptotic DNA fragments significantly increased in the experimental group, and ladder phenomenon appeared. Conclusion: DMY can inhibit proliferation and promote apoptosis of lung cancer A549 cells, which may be related to the breakage of chromosome DNA.

关键词(KeyWords): 二氢杨梅素;肺肿瘤;癌;细胞增殖;细胞凋亡;DNA ladder凝胶电泳
dihydromyricetin;lung neoplasms;carcinoma;cell proliferation;cell apoptosis;DNA ladder gel electrophoresis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31360215);; 贵州省教育厅青年科技人才成长项目[黔教合KY字(2018)177];; 中国医学科学院中央级公益性科研院所基本科研业务费专项资金(2017PT31042,2018PT31048)

作者(Authors): 刘金河;葛章文;张望明;晏文涛;刘志;吴昌学;
LIU Jinhe;GE Zhangwen;ZHANG Wangming;YAN Wentao;LIU Zhi;WU Changxue;National & Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Center of Tissue Engineering and Stem Cell Research, Guizhou Provincial Key Laboratory for Regenerative Medicine, Guizhou Medicine University;Key Laboratory of Adult Stem Cell Translational Research, Chinese Academy of Medical Sciences;Department of Laboratory Medicine, Guizhou Provincial People's Hospital;The First Affiliated Hospita

DOI: 10.19367/j.cnki.1000-2707.2019.10.005

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