姜黄素对小鼠肝癌细胞体外增殖和凋亡的影响  

Effects of curcumin on in vitro proliferation and apoptosis of mouse hepatocellular carcinoma cells

作　　者：贾红敏 王欣欣 张斌[1] JIA Hongmin;WANG Xinxin;ZHANG Bin(Department of Pharmacy,The Second Hospital of Shandong University,Jinan 250031,China)

机构地区：[1]山东大学第二医院药学部,济南250031

出　　处：《山东医药》2024年第16期42-46,共5页Shandong Medical Journal

摘　　要：目的探讨姜黄素对小鼠肝癌Hepa1-6细胞体外增殖和凋亡的影响。方法取处于对数生长期的Hepa1-6细胞,随机分为对照组、10μmol/L组、20μmol/L组和40μmol/L组,分别采用0、10、20、40μmol/L的姜黄素进行干预,通过MTT法检测细胞增殖,流式细胞术检测细胞凋亡。RT-qPCR法检测Hepa1-6细胞中凋亡相关分子Bcl-2和Caspase-3及Toll样受体2(TLR2)、IFN-α、IFN-β基因表达。ELISA法检测Hepa1-6细胞培养上清IFN-Ⅰ。对照组、40μmol/L组中Hepa1-6细胞分别与CD4+T细胞、CD8+T细胞和NK细胞共孵育,流式细胞术检测CD4+T细胞、CD8+T细胞和NK细胞CD69表达。结果10、20、40μmol/L组Hepa1-6细胞增殖抑制率呈剂量依赖性增加,抑制率依次为(36.45±2.11)%、(46.40±2.08)%、(82.16±3.32)%,与对照组比较差异均有统计学意义(P均<0.05);10、20和40μmol/L组Hepa1-6细胞凋亡比例分别为10.47%、26.66%和49.81%,与对照组比较差异均有统计学意义(P均<0.01)。RT-qPCR结果显示,10、20和40μmol/L组中抗凋亡基因Bcl-2转录水平分别是对照组的(0.77±0.06)倍、(0.44±0.06)倍和(0.25±0.08)倍,与对照组比较差异均有统计学意义(P均<0.01);同时,促凋亡基因Caspase-3表达增高,10、20和40μmol/L组中促凋亡基因Caspase-3转录水平分别是对照组的(1.73±0.31)倍、(2.52±0.45)倍和(3.25±0.20)倍,与对照组比较差异均有统计学意义(P均<0.01)。随姜黄素浓度的增加,TLR2转录水平逐渐升高,10、20和40μmol/L组TLR2转录水平分别是对照组的(3.56±0.59)倍、(4.93±0.04)倍和(6.60±0.47)倍,与对照组比较差异均有统计学意义(P均<0.05);10、20和40μmol/L组IFN-α转录水平升高,分别为对照组的(4.21±0.22)倍、(5.54±0.39)倍和(8.05±0.93)倍,与对照组比较差异均有统计学意义(P均<0.05);IFN-β转录水平分别为对照组的(3.08±0.51)倍、(5.31±0.55)倍和(8.35±0.59)倍,与对照组比较差异均有统计学意义(P均<0.01)。10、20和40μmol/L组细胞分泌IFN-α水平�Objective To explore the effects of curcumin on the in vitro proliferation and apoptosis of mouse liver can-cer Hepa1-6 cells.Methods Hepa1-6 cells in the logarithmic growth phase were divided into the control group,10μmol/L group,20μmol/L group and 40μmol/L group,respectively,which were treated with 0,10,20,and 40μmol/L of cur-cumin.Cell proliferation was assessed by the MTT assay,and apoptosis level was detected by FACS.Real-time quantitative PCR(RT-qPCR)was employed to detect the levels of apoptosis-related molecules Bcl-2,Caspase-3,Toll-like receptor 2(TLR2),IFN-α,and IFN-βin Hepa1-6 cells.The secretion levels of IFN-αand IFN-βin the cell culture supernatant were determined using ELISA.Hepa1-6 cells were incubated with CD4+T cells,CD8+T cells and NK cells in the control group and 40μmol/L curcumin group,respectively.FACS was used to detect the expression levels of CD69 in CD4+T cells,CD8+T cells and NK cells.Results The proliferation inhibition rates of Hepa1-6 cells in the 10,20 and 40μmol/L groups increased in a dose-dependent manner,and the inhibition rates were(36.45±2.11)%,(46.40±2.08)%and(82.16±3.32)%,respectively.There was statistically significant difference in comparison with that of the control group(P<0.05).The apoptotic proportions of Hepa1-6 cells in the 10μmol/L,20μmol/L and 40μmol/L groups were 10.47%,26.66%and 49.81%,respectively.And the difference was statistically significant in comparison with that of the control group(P<0.01).RT-qPCR results showed that the transcription levels of anti-apoptotic gene Bcl-2 in the 10μmol/L,20μmol/L and 40μmol/L groups were(0.77±0.06)times,(0.44±0.06)times and(0.25±0.08)times that of the control group,respectively,with statistically significant difference(all P<0.01).At the same time,the expression of pro-apoptotic gene Caspase-3 significantly increased,and the transcription levels of pro-apoptotic gene Caspase-3 in the 10μmol/L,20μmol/L and 40μmol/L groups was(1.73±0.31)times,(2.52±0.45)times and(3.25±0.20)times that of the contr

关 键 词：肝癌 姜黄素 TOLL样受体2 细胞增殖 细胞凋亡 免疫功能 

分 类 号：R735.7[医药卫生—肿瘤]