返回 
加入收藏
蓝萼甲素通过Bcl-2/Beclin1靶点调节人肝癌HCCLM3细胞自噬与凋亡的机制研究
x
请在关注微信后，向客服人员索取文件
| 篇名： | 蓝萼甲素通过Bcl-2/Beclin1靶点调节人肝癌HCCLM3细胞自噬与凋亡的机制研究 |
| TITLE： | Study on regulation mechanism of glaucocalyxin A on the autophagy and apoptosis of HCCLM 3 hepatocellular carcinoma cells through Bcl- 2/Beclin1 target |
| 摘要： | 目的 研究蓝萼甲素（GLA）对人肝癌HCCLM3细胞自噬与凋亡的调节作用机制。方法HCCLM3细胞按不同实验目的主要设对照组、GLA2.5μg/mL组、GLA5μg/mL组、GLA10μg/mL组。对照组仅加入完全培养基，各给药组分别加入含GLA相应终质量浓度的完全培养基。采用流式细胞术检测细胞周期分布与凋亡情况；采用透射电镜观察细胞线粒体形态和自噬情况（仅设对照组、GLA5μg/mL组）；采用JC-1染色和荧光倒置显微镜观察并检测细胞线粒体膜电位；采用Westernblot法检测细胞中Bcl-2、Bax、Beclin1、活性半胱氨酸天冬氨酸蛋白酶3（cleavedcaspase-3）蛋白的表达；采用免疫共沉淀法检测细胞中Bcl-2与Beclin1的结合与解离（仅设GLA5μg/mL组、GLA10μg/mL组）。结果与对照组相比，GLA5、10μg/mL能够诱发细胞周期显著阻滞于G2～M期、诱导线粒体膜电位显著下降、增加促凋亡作用，还能显著促进Bax、cleavedcaspase-3、Beclin1蛋白表达，显著抑制Bcl-2蛋白表达（P＜0.01）；GLA5μg/mL还可引发线粒体形态显著改变，自噬小体增多。免疫共沉淀法结果显示，与GLA5μg/mL比较，GLA10μg/mL能够增强Bcl-2与Beclin1的结合。结论GLA可能通过Bcl-2/Beclin1靶点调节HCCLM3细胞自噬与凋亡，且作用效果与剂量密切相关。 |
| ABSTRACT： | OBJECTIVE To study the regulator y mech anism of glaucocalyxin A （GLA） on autophagy and apoptosis of HCCLM3 hepatocellular carcinoma cells. METHODS HCCLM3 cells were taken ，and control group ，GLA 2.5 μg/mL group，GLA 5 μg/mL group and GLA 10 μg/mL group were mainly set according to different experimental purposes. In control group，only complete medium was added ；in each administration group ，complete medium containing the corresponding final concentration of GLA was added. Cell cycle distribution and apoptosis were detected by flow cytometry ；mitochondrial morphology and autophagy were observed by transmission electron microscope （only control group ，GLA 5 μg/mL group）；JC-1 staining and fluorescence inverted microscope were used to observe and detect the mitochondrial membrane potential of the cells ；Western blot assay was used to detect the protein expression of Bcl- 2， Bax， Beclin1 and cleaved caspase- 3 proteins in the cells ； the co-immunoprecipitation method was used to detect the binding and dissociation of Bcl- 2 and Beclin 1（only GLA 5 μg/mL group， GLA 10 μg/mL group）. RESULTS Compared with control group ，GLA 5 μg/mL and GLA 10 μg/mL could induce a significant arrest of the cell cycle in the G 2-M phase for HCCLM 3，a significant decrease in mitochondrial membrane potential ，an increase in apoptosis as well as significant promotion of the protein expression of Bax ，cleaved caspase- 3 and Beclin 1，and significant inhibition of the protein expression of Bcl- 2（P＜0.01）. GLA 5 μg/mL also significantly changed mitochondrial morphology and increased autophagosomes. The results of co-immunoprecipitation showed that compared with GLA 5 μg/mL，GLA 10 μg/mL could enhance the binding of Bcl- 2 and Beclin 1. CONCLUSIONS GLA can regulate the autophagy and apoptosis of HCCLM 3 cells by Bcl-2/Beclin1 target. The effect is closely related to the dose of GLA. |
| 期刊： | 2022年第33卷第09期 |
| 作者： | 朱琳琳，张明明，郭格格，徐祉轩 |
| AUTHORS： | ZHU Linlin，ZHANG Mingming，GUO Gege，XU Zhixuan |
| 关键字： | 蓝萼甲素；靶点；机制；自噬；凋亡；Bcl-2；Beclin1 |
| KEYWORDS： | glaucocalyxin A ；target；mechanism；autophagy；apoptosis；Bcl-2；Beclin1 |
| 阅读数： | 240 次 |
| 本月下载数： | 8 次 |
* 注：未经本站明确许可，任何网站不得非法盗链资源下载连接及抄袭本站原创内容资源!在此感谢您的支持与合作！
