摘要
目的:探究青蒿琥酯(ART)对人胃癌细胞增殖、凋亡和自噬以及激活转导因子4/C/EBP同源蛋白(ATF4/CHOP)通路的影响。方法:细胞计数试剂盒8(CCK-8)法和乳酸脱氢酶(LDH)法筛选ART干预人胃癌细胞系MGC803的最佳条件。原位末端标记(TUNEL)染色法观察细胞凋亡,并对半胱氨酸天冬氨酸蛋白水解酶(Caspase)-3和Caspase-9活性进行测定;免疫荧光染色法观察细胞自噬蛋白微管相关蛋白轻链3(LC3)表达;Western Blot分析验证自噬(Beclin和ATG5)和ATF4/CHOP通路相关蛋白表达。结果:ART以浓度依赖性方式降低MGC803细胞活力(P<0.05)。ART治疗后促进MGC803细胞凋亡、Caspase-3和Caspase-9活性,增强LC3、Beclin和ATG5表达,上调ATF4和CHOP表达(P<0.05)。自噬抑制剂3-MA或沉默ATF4可部分逆转ART对胃癌细胞的影响(P<0.05)。结论:ART可能通过激活ATF4/CHOP通路在胃癌细胞中发挥抗增殖和促凋亡、自噬作用。
Objective: To investigate the impacts of artesunate(ART) on the proliferation, apoptosis, autophagy and ATF4/CHOP pathway of human gastric cancer cells.Methods: CCK-8 method and lactate dehydrogenase(LDH) method were used to screen the optimal conditions for ART intervention in human gastric cancer cell line MGC803. Apoptosis was observed by TUNEL staining, and the activities of Caspase-3 and Caspase-9 were measured;the autophagy protein LC3 was observed by immunofluorescence staining, and the expression of autophagy-related proteins(Beclin and ATG5) and ATF4/CHOP pathway-related proteins was verified by Western blot.Results: ART decreased MGC803 cell viability in a concentration-dependent manner(all P<0.05). ART treatment promoted MGC803 cell apoptosis, and Caspase-3 and Caspase-9 activities, enhanced LC3, Beclin and ATG5 expression, and up-regulated ATF4 and CHOP expression(all P<0.05). Autophagy inhibitor 3-MA or silencing of ATF4 could partially reverse the effects of ART on gastric cancer cells(all P<0.05). Conclusion: ART may exhibit anti-proliferative, pro-apoptotic and autophagic effects in gastric cancer cells by activating ATF4/CHOP pathway.
作者
马立东
刘晓蕾
赵飞
Ma Lidong;Liu Xiaolei;Zhao Fei(First Department of Gastroenterology,First Hospital of Handan City,Hebei Handan 056000,China;First Department of Geriatrics,First Hospital of Handan City;Department of Gastroenterology,Hebei Youfu Hospital)
出处
《中国药师》
CAS
2022年第11期1884-1888,共5页
China Pharmacist
基金
河北省医学科学课题研究计划项目(编号:20220491)。
