吉林大学学报（医学版）2018，Vol.44Issue(1)：52-57,6.DOI:10.13481/j.1671-587x.20180110
咖啡因联合电离辐射对沉默Chk-1肝癌干细胞的增殖抑制和凋亡诱导作用
Inhibitory effect of caffeine combined with ionizing radiation on proliferation of hepatocellular carcinoma stem cells silenced by Chk-1 and its apoptosis-induced effect
摘要
Abstract
Objective:To treat the hepatocellular carcinoma stem cells silenced by Chk-1 with caffeine combined with 4 Gy X-ray irradiaition,and to explore their synergistic killing effects on the hepatocellular carcinoma stem cells by detecting the cell proliferation,cell cycle and apoptosis.Methods:The lentivirus silencing Chk-1 was transfected into the 293T cells;after the HepG2 cells were infected by the lentivirus,the Chk-1 protein expression was detected by Western blotting to confirm the silencing efficency,and non-target control was established,and HepG2-Chk-1 and HepG2-control were named.The cells highly expressed CD133 were cultured by suspension culture method,S-HepG2-Chk-1 and S-HepG2-control were named respectively.After the cells were treated by caffeine,they were irradiated by 4 Gy X-ray;the proliferation activity was measured by MTT,and the cell cycle distribution and the apoptotic rate were measured by PI single staining and AnnexinⅤ-FITC double staining using FCM,respectively.For proliferation,cell cycle and apoptosis assays,there were control,caffeine,4 Gy and caffeine+4 Gy groups.Results:The Western blotting results showed that after the HepG2 cells were infected by lentivirus,the Chk-1 protein expression was significantly decreased,but it was not obvious in non-target control group,it demonstrated that the cell models HepG2-Chk-1 and HepG2-control were obtained successfully.After the HepG2-Chk-1 and HepG2-control cells were cultivated by suspension,the CD133 protein expression were increased,it demonstrated that there were high proportion of CD133+ cells,they were hepatocellular carcinoma stem cells.Compared with control group,the proliferation activities in caffeine group and 4 Gy group were significantly decreased (P＜0.05 or P＜0.01),the percentages of cells at G2/M phage and the apoptotic rates were significantly increased (P＜0.05 or P＜0.01),and the percentage of cells at S phage in caffeine group was significantly increased (P＜0.05);the percentage of cells at G0/G1 phage in 4 Gy group was increased (P＜0.05 or P＜0.01),the effect was more stronger in caffeine+4 Gy group.Compared with S-HepG2-control,the proliferation activities of S-HepG2-Chk-1 ceils in caffeine group and 4 Gy group were decreased,the apoptotic rates were increased (P＜0.05 or P＜0.01),and the percentage of cells at G1/M phage was significantly decreased (P＜0.05 or P＜0.01).Collusion:The hepatoccellular carcinoma stem cells silenced by Chk-1 with positive CD133 are obtained successfully;Caffeine combined with X-ray irradiation can inhibit the cell proliferation and induce the apoptosis,and enhance the G2/M arrest,and both of them have synergistic effects.关键词
咖啡因/X射线/肝肿瘤/肿瘤干细胞/细胞增殖/细胞凋亡Key words
caffeine/X-ray/liver neoplasms/cancer stem cell/cell proliferation/apoptosis分类
生物科学引用本文复制引用
于雷,王志成,王铁君..咖啡因联合电离辐射对沉默Chk-1肝癌干细胞的增殖抑制和凋亡诱导作用[J].吉林大学学报（医学版）,2018,44(1):52-57,6.基金项目
吉林省卫计委科研计划项目资助课题(2015Z009) （2015Z009）
吉林省教育厅科学技术项目资助课题(JJKH20170828KJ) （JJKH20170828KJ）
