没食子酸对人食管癌TE-1细胞的体外抑制作用及其机制  

Inhibitory effects of gallic acid on human esophageal cancer TE-1 cells in vitro and its mechanism

作　　者：吴昊 努兰·拜都拉 刘琳玉 任艳利 WU Hao;Nuran·Baydolla;LIU Linyu;REN Yanli(College of Biology and Geography,Yili Normal University,Xinjiang Yining 835000,China;Key Laboratory for Protection,Development and Utilization of Biological Resources,Yili Normal University,Xinjiang Yining 835000,China)

机构地区：[1]伊犁师范大学生物与地理科学学院,新疆伊宁835000 [2]伊犁师范大学微生物资源保护与开发利用重点实验室,新疆伊宁835000

出　　处：《中国药房》2022年第12期1448-1454,共7页China Pharmacy

基　　金：新疆维吾尔自治区研究生科研创新计划项目(No.XJ2020G303)。

摘　　要：目的探究没食子酸(GA)对人食管癌TE-1细胞的体外抑制作用及潜在机制。方法MTT法检测GA作用24 h和48 h后对TE-1细胞增殖的影响,细胞荧光计数(CCK-F)法和倒置荧光显微镜观察GA作用后TE-1细胞的数量变化和形态变化,划痕实验检测细胞迁移能力的变化,平板集落形成实验检测GA对TE-1细胞集落形成能力的影响,流式细胞术检测细胞凋亡情况,荧光探针(DCFH-DA)法观察活性氧(ROS)产生情况;Western blot法检测胱天蛋白酶3(caspase-3)、caspase-9、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)和细胞周期蛋白D1(cyclin D1)、cyclin D3的表达水平。结果GA能够明显降低TE-1细胞的增殖能力,且有时间和浓度依赖趋势,其对细胞的半数抑制浓度分别为(281.22±26.81)μmol/L(24 h)和(220.90±31.15)μmol/L(48 h)。与对照组比较,给药组细胞出现皱缩、排列稀疏、细胞核固缩等现象,且细胞数量明显减少。与对照组比较,给药组细胞的细胞迁移能力和集落形成能力均显著降低(P<0.01或P<0.05)。经100、300、500μmol/L的GA作用24 h后,细胞凋亡率分别为(6.21±0.32)%、(12.59±0.58)%和(15.41±0.41)%,均显著高于对照组的(5.29±0.28)%(P<0.01或P<0.05);除100μmol/L GA组外,其余给药组细胞ROS的产生水平均显著提高(P<0.01或P<0.05);与对照组比较,各给药组细胞中Bcl-2(仅GA 200μmol/L组)、Bax(GA 100μmol/L组除外)、caspase-3、caspase-9(GA 100μmol/L组除外)的表达水平均显著升高(P<0.01或P<0.05),Bcl-2(GA 100、200μmol/L组除外)、cyclin D1、cyclin D3蛋白的表达水平均显著降低(P<0.01或P<0.05)。结论GA能够抑制人食管癌TE-1细胞的增殖,限制其迁移能力及集落形成能力,促进其凋亡;上述作用可能与该成分可提高ROS水平并上调促凋亡蛋白caspase-3、caspase-9、Bax的表达,下调抗凋亡蛋白Bcl-2和细胞周期蛋白cyclin D1、cyclin D3的表达有关。OBJECTIVE To investigate inhibitory effects of gallic acid(GA)on human esophageal cancer TE-1 cells in vitro and its potential mechanism. METHODS The effects of GA on the proliferation of TE-1 cells were determined by MTT assay after treated with GA for 24 h and 48 h. Cell fluorescence counting(CCK-F)method and inverted fluorescence microscope were used to observe the changes in the number and morphology of TE-1 cells after treated with GA. The change of cell migration ability was detected by scratch test. The effects of GA on the colony-forming ability of TE-1 cells were tested by plate colony formation experiment. Cell apoptosis was detected by flow cytometry. Fluorescence probe(DCFH-DA)method was used to observe reactive oxygen species(ROS) production. Western blot assay was used to detect the expression of caspase-3,caspase-9,Bcl-2,Bcl-2 associated X protein(Bax),cyclin D1 and cyclin D3. RESULTS GA significantly reduced the proliferation ability of TE-1 cells in time and concentration dependent manner. the IC50 of GA to TE-1 cells were(281.22±26.81)μmol/L(24 h)and(220.90±31.15)μmol/L(48 h),respectively. Compared with control group,the cells in the administration group showed shrinkage,sparse arrangement and nuclear pyknosis,and the number of cells decreased significantly. Compared with control group,the cell migration ability and colony formation ability were decreased significantly in administration groups(P<0.01 or P<0.05). The apoptosis rates of TE-1 cells were(6.21±0.32)%,(12.59±0.58)% and(15.41±0.41)% after treated with 100,300 and 500 μmol/L GA for 24 h,all of which were significantly higher than(5.29±0.28)% of control group(P<0.01 or P<0.05). Except for GA 100 μmol/L group,the level of ROS in other administration groups were significantly increased(P<0.01 or P<0.05). Compared with control group,the expressions of Bcl-2(only GA 200 μmol/L group),Bax(except for GA 100 μmol/L),caspase-3 and caspase-9(except for GA 100 μmol/L)were increased significantly(P<0.01 or P<0.05),while the protein exp

关 键 词：没食子酸 食管癌 凋亡 活性氧 TE-1细胞 

分 类 号：R963[医药卫生—微生物与生化药学]