摘要
目的:体外实验探讨茶多酚(TP)联合奥沙利铂(L-OHP)对人胃癌细胞化疗敏感性的作用及其机制。方法:选取人胃癌HGC-27和N87细胞及人胃黏膜GES-1细胞,采用CCK-8实验测定TP和L-OHP分别处理的HGC-27和N87细胞活力,并计算L-OHP的IC_(50),测定L-OHP联合TP处理的GES-1细胞活力。将HGC-27和N87细胞随机分为对照组、TP(5μmol/L)组、L-OHP(HGC-27细胞5.2μmol/L,N87细胞7.7μmol/L)组和TP(5μmol/L)+LOHP(HGC-27细胞5.2μmol/L,N87细胞7.7μmol/L)组,每组3个复孔。CCK-8实验测定不同处理组的细胞活力;集落形成实验检测细胞增殖能力;划痕实验和Transwell实验检测细胞迁移和侵袭能力;流式细胞术检测细胞凋亡;腺病毒感染检测细胞自噬;活性氧(ROS)检测试剂盒检测ROS水平;Western blot检测微管相关蛋白1轻链3(LC3)、核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)和超氧化物歧化酶1(SOD1)蛋白水平。结果:与L-OHP单独处理相比,TP联合L-OHP处理后HGC-27和N87细胞的活力下降显著,细胞增殖和迁移能力受到显著抑制,自噬小体和ROS水平显著升高,LC3-II/LC3-I比值显著升高,Nrf2、HO-1和SOD1表达水平显著降低(P<0.05)。结论:TP通过抑制Nrf2通路、再生细胞内ROS促进细胞自噬,从而提高胃癌细胞对L-OHP的敏感性。
AIM:To investigate the effect of tea polyphenols(TP)on the sensitivity of human gastric cancer cells to oxaliplatin(L-OHP)in vitro and its mechanism.METHODS:Human gastric cancer HGC-27 and N87 cells,and human gastric mucosal GES-1 cells were used in this study.The HGC-27 and N87 cells were randomly assigned into control group,TP(5μmol/L)group,L-OHP(5.2μmol/L for HGC-27 cells,7.7μmol/L for N87 cells)group,and TP(5μmol/L)combined with L-OHP(5.2μmol/L for HGC-27 cells,7.7μmol/L for N87 cells)group,with 3 replicate wells per group.The cell viability was detected by CCK-8 assay,and the IC_(50) value of L-OHP was calculated.The proliferation of the cells was assessed by colony formation assay.The migration and invasion abilities of the cells were evaluated by scratch test and Transwell assay.Flow cytometry was performed to evaluate the antiapoptotic effect of the treatments.Adenovirus infection was conducted to evaluate cell autophagy.The levels of intracellular reactive oxygen species(ROS)were assessed using a ROS assay kit.Western blot analysis was performed to evaluate the protein expression levels of microtubule-associated protein 1 light chain 3(LC3),nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1)and superoxide dismutase 1(SOD1).RESULTS:Combination of TP and L-OHP significantly reduced the viability of HGC-27 and N87 cells,and markedly inhibited cell proliferation and migration compared with L-OHP alone.Significant increases in autophagosomes and ROS levels were observed in combination group compared with L-OHP alone group.The ratio of LC3-II/LC3-I significantly increased in combination group,whereas the expression of Nrf2,HO-1 and SOD1 significantly decreased compared with L-OHP alone group(P<0.05).CONCLUSION:Treatment with TP enhanced the sensitivity of HGC-27 and N87 cells to L-OHP by inhibiting the Nrf2 pathway,promoting the production of intracellular ROS,and inducing cell autophagy.
作者
董钦鹏
孙晓东
刘君瑞
王鑫澳
刘家铭
李恒平
DONG Qinpeng;SUN Xiaodong;LIU Junrui;WANG Xinao;LIU Jiaming;LI Hengping(Department of General Surgery,Jinzhou Medical University&Xiangyang No.1 People's Hospital Union Training Base,Xiangyang 441000,China;Translational Medicine Center,Xiangyang No.1 People's Hospital Affiliated to Hubei Univer-sity of Medicine,Xiangyang 441000,China;Department of General Surgery,Xiangyang No.1 People's Hospital Affiliatedto Hubei University of Medicine,Xiangyang 441000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2024年第7期1182-1189,共8页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81703015)
湖北省科技计划项目(No.2021CFB582)。
关键词
胃癌
茶多酚
奥沙利铂
自噬
活性氧
gastric cancer
tea polyphenols
oxaliplatin
autophagy
reactive oxygen species
