摘要
目的研究腺苷对肝癌细胞增殖和自噬的作用。方法腺苷作用于HepG2细胞,CCK-8法观察细胞增殖的变化,Western blot研究LC3-Ⅱ和LC3-Ⅰ蛋白的表达,MDC染色观察自噬体数量。结果腺苷(1.0~4.0 mmol·L^(-1))作用于HepG2细胞,在12、24、48 h可明显抑制细胞增殖(P<0.01)。低浓度腺苷(0.2、0.5、1.0 mmol·L^(-1))处理细胞24h,LC3-Ⅱ/LC3-Ⅰ明显降低,腺苷(1.0 mmol·L^(-1))对HepG2细胞自噬抑制作用最强(P<0.01),高浓度腺苷(4 mmol·L^(-1))反而增强Hep G2细胞内自噬(P<0.05)。腺苷(1.0mmol·L^(-1))作用细胞24 h,LC3-Ⅱ/LC3-Ⅰ比值在6、12、24 h检测点明显降低,MDC染色显示自噬体减少,在12 h检测点发现LC3-Ⅱ/LC3-Ⅰ最低(P<0.01),自噬体基本消失。结论腺苷抑制肝癌细胞增殖,低浓度腺苷抑制细胞自噬,高浓度腺苷促进细胞自噬,对提高肝癌药物的治疗效果有重要意义。
Aim To investigate the effect of adenosine on the autophagy and proliferation of hepatocellular carcinoma cells,and improve the curative effect of adenosine on hepatocellular carcinoma.Methods HepG2 cells were incubated with adenosine,CCK-8 method was used to study the changes of cell proliferation,Western blot was used to study the expression of LC3-Ⅱand LC3-Ⅰ,and MDC staining was used to observe the number of autophagosomes.Results HepG2 cells were incubated with adenosine(1.0~4.0 mmol·L-1)for 48 h,the proliferation of HepG2 cells were detected at the different time points(12,24,48 h),and the result showed the proliferation was significantly inhibited by adenosine(P<0.01).HepG2 cells were incubated with adenosine(0.2,0.5,1.0,2.0,4.0 mmol·L-1)for 24 h,the ratio of LC3-Ⅱ/LC3-Ⅰdecreased significantly in low concentration of adenosine group(0.2,0.5 mmol·L-1,P<0.05;1.0 mmol·L-1,P<0.01),and the ratio of LC3-Ⅱ/LC3-Ⅰincreased significantly in higher concentration of adenosine group(4.0 mmol·L-1,P<0.05).HepG2 cells were incubated with adenosine(1.0 mmol·L-1)for 24 h,the ratio of LC3-Ⅱ/LC3-Ⅰdecreased significantly at 6,12 and 24 h detecting point,the number of autophagosomes were reduced,the lowest ratio of LC3-Ⅱ/LC3-Ⅰand autophagosomes were observed at 12 h detecting point(P<0.01).Conclusions Adenosine inhibits the proliferation of hepatocellular carcinoma cells,the low concentration of adenosine inhibits the autophagy,while the high concentration of adenosine increases the autophagy,which is of great significance to reduce multi-drug resistance and improve the therapeutic effect of anti-hepatoma drugs.
作者
蒲泽锦
周小涛
李国平
詹浩炼
郭益添
项梦琦
刘丽璇
谭辉
吴灵飞
PU Ze-jin;ZHOU Xiao-tao;LI Guo-ping;ZHAN Hao-lian;GUO Yi-tian;XIANG Meng-qi;LIU Li-xuan;Tan Hui;WU Ling-fei(Dept of Gastroenterology,the Second Affiliated Hospital of Shantou University Medical College,Shantou,Guangdong 515041,China;Dept of Radiology,the Second Affiliated Hospital of Shantou University Medical College,Shantou,Guangdong 515041,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2018年第4期508-512,共5页
Chinese Pharmacological Bulletin
基金
广东省科技计划资助项目(No 2014A020212284)
广东省自然科学基金资助项目(No 2014A030313470)
关键词
腺苷
自噬
多药耐药
肝癌
细胞增殖
肿瘤治疗
adenosine
autophagy
multi-drug resistance
hepatocellular carcinoma
cell proliferation
tumor therapy