槲皮素经抑制己糖激酶活性介导人肝癌Huh-7细胞的增殖、侵袭和转移  

Quercetin mediates the proliferation,invasion and metastasis of human hepatoma Huh-7 cells via inhibition of hexokinase activity

作　　者：林丽彬[1,2] 谢群 邱新悦 吴梦羽 徐丽 张静霞[1,2] Lin Libin;Xie Qun;Qiu Xinyue;Wu Mengyu;Xu Li;Zhang Jingxia(School of Basic Medicine,Putian University,Putian 351100,China;Key Laboratory of Translational Tumor Medicine in Fujian Province,Putian University,Putian 351100,China)

机构地区：[1]莆田学院基础医学院,莆田351100 [2]肿瘤转化医学福建省高校重点实验室,莆田351100

出　　处：《中华实验外科杂志》2023年第10期1951-1954,共4页Chinese Journal of Experimental Surgery

基　　金：莆田市科技计划项目(2019SP004)。

摘　　要：目的探讨槲皮素对人肝癌Huh-7细胞生物学行为及其己糖激酶(Hexokinase)活性的影响。方法0μmol/L(对照组)、100、150、200μmol/L槲皮素处理Huh-7细胞,24、48、72 h后采用细胞计数试剂盒(CCK-8)检测Huh-7细胞的增殖能力,24 h后采用Transwell法评估Huh-7细胞的迁移及侵袭能力;24 h后应用蛋白质印迹法(Western blot)及实时定量反转录聚合酶链反应(RT-qPCR)法检测Huh-7细胞己糖激酶mRNA转录及蛋白水平表达变化;24 h后采用己糖激酶检测试剂盒检测细胞己糖激酶的活性改变。多组定量资料差异比较采用方差分析,不同槲皮素浓度组的各个指标数值与空白对照组数值的均数间多重比较采用Dunnett法。结果3种浓度的槲皮素处理Huh-7细胞增殖抑制率均高于对照组(24 h:38.438±6.787、45.776±5.463、56.376±6.457比0.000±0.000,F=102.593,P<0.001;48 h:56.395±6.503、75.124±10.216、83.943±6.297比0.000±0.000,F=152.597,P<0.001;72 h:71.845±8.267、86.331±7.139、93.024±4.072比0.000±0.000,F=269.481,P<0.001)。3种浓度的槲皮素处理组的细胞迁移(277.000±34.400、228.533±34.734、183.400±32.300比375.333±39.233,F=16.331,P<0.001)和侵袭(36.000±13.440、18.600±4.850、13.733±4.388比199.667±4.163,F=394.462,P<0.001)能力均明显低于对照组。己糖激酶的mRNA转录(0.650±0.137、0.378±0.204、0.164±0.046比1.000±0.000,F=25.051,P<0.001)及蛋白(0.615±0.128、0.540±0.125、0.480±0.138比1.000±0.000,F=12.863,P<0.01)水平均明显低于对照组,己糖激酶的活性明显低于对照组(0.306±0.064、0.247±0.090、0.143±0.037比0.535±0.142,F=9.768,P<0.01),差异均有统计学意义。结论槲皮素可通过抑制己糖激酶mRNA转录及蛋白表达水平,并降低己糖激酶活性来抑制人肝癌Huh-7细胞的增殖、侵袭和转移。Objective To explore the effect of quercetin on the biological behavior of human hepatoma Huh-7 cells and its hexokinase HK activity.Methods Huh-7 cells were treated with 0μmol/L(control),100,150,200μmol/L quercetin,and the proliferative ability of Huh-7 cells was detected by cell counting kit-8(CCK-8)assay after 24,48 and 72 h,The migratory and invasive ability of Huh-7 cells was assessed by Transwell assay after 24 h;The changes in the expression of mRNA transcript and protein levels of hexokinase were detected by Western blotting and real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)assays 24 h later;The activity of cellular hexokinase was detected by using hexokinase assay kit after 24 h.Comparison of differences in quantitative data of multiple groups was performed by ANOVA,and multiple comparisons between the means of the values of each index in different quercetin concentration groups and the values of the blank control group were performed by Dunnett’s method.Results The proliferation inhibition rate of Huh-7 cells treated with three concentrations of quercetin was higher than that of the control group(24 h:38.438±6.787,45.776±5.463,56.376±6.457 vs.0.000±0.000,F=102.593,P<0.001;48 h:56.395±6.503,75.124±10.216,83.943±6.297 vs.0.000±0.000,F=152.597,P<0.001;72 h:71.845±8.267,86.331±7.139,93.024±4.072 vs.0.000±0.000,F=269.481,P<0.001).Compared with the control group,the three concentrations of quercetin-treated groups showed higher cell migration(277.000±34.400,228.533±34.734,183.400±32.300 vs.375.333±39.233,F=16.331,P<0.001)and invasion(36.000±13.440,18.600±4.850,13.733±4.388 vs.199.667±4.163,F=394.462,P<0.001)abilities were significantly reduced.The mRNA transcription(0.650±0.137,0.378±0.204,0.164±0.046 vs.1.000±0.000,F=25.051,P<0.001)and protein(0.615±0.128,0.540±0.125,0.480±0.138 vs.1.000±0.000,F=12.863,P<0.01)levels were significantly decreased,and the activity of hexokinase was significantly decreased(0.306±0.064,0.247±0.090,0.143±0.037 vs.

关 键 词：肝癌 槲皮素 细胞生物学行为 己糖激酶 

分 类 号：R285[医药卫生—中药学]