高菜硫代葡萄糖苷提取物对人结肠癌细胞HCT116的抑制作用  

Inhibitory Effect of Glucosinolates from Brassica juncea var.integlifolia on HCT116 Human Colon Cancer Cells

作　　者：田艳[1] 邓放明[1] 赵玲艳[1] 廖安[1] 赖灯妮 陈秋佳 TIAN Yan;DENG Fangming;ZHAO Lingyan;LIAO An;LAI Dengni;CHEN Qiujia(College of Food Science and Technology,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学食品科学技术学院,湖南长沙410128

出　　处：《食品科学》2020年第23期172-180,共9页Food Science

基　　金：国家现代农业产业技术体系建设专项(CARS-24-E-02);湖南农业大学双一流建设项目(SYL201802006)。

摘　　要：为了探讨高菜(Brassica juncea var.integlifolia)中硫代葡萄糖苷(简称硫苷)的含量、种类及其对人结肠癌细胞株HCT116细胞增殖和凋亡的影响,采用紫外分光光度法检测新鲜高菜中总硫苷含量,高效液相色谱-电喷雾质谱联用技术鉴定硫苷化合物结构,流式细胞仪、蛋白质印迹法和实时定量聚合酶链式反应检测高菜硫苷提取物对人结肠癌细胞株HCT116细胞增殖和凋亡的影响。结果表明:新鲜高菜中总硫苷含量为15.45 mg/g;从高菜硫苷提取物(总硫苷质量分数为73.7%)中鉴定出2-丙烯基硫苷、4-甲基硫代丁基硫苷、4-甲氧基-3-吲哚甲基硫苷3种主要的硫苷化合物;高菜硫苷提取物对正常人结肠成肌纤维细胞CCD-18Co没有抑制作用,但通过诱导细胞周期基因及相关信号蛋白(Cyclin B、CyclinD1、CyclinE)下调使HCT116细胞的细胞周期停滞,通过诱导细胞凋亡基因及相关信号蛋白(Caspase-3、Cleaved caspase-3)的上调使人结肠癌细胞株HCT116细胞凋亡,从而抑制人结肠癌细胞株HCT116细胞增殖。In this study,we determined the content of total glucosinolates in fresh Brassica juncea var.integlifolia by UV spectrophotometry,identified the structures of glucosinolates by high performance liquid chromatography of quadrupole time of flight-tandem mass spectrometry(HPLC-Q-TOF-MS),and evaluated the effects of glucosinolates extracted from Brassica juncea var.integlifolia on the proliferation and apoptosis of human colon cancer cell line HCT116 by flow cytometry,Western blot and quantitative real-time PCR.The results showed that the content of total glucosinolates in fresh Brassica juncea var.integlifolia was 15.45 mg/g.Three major glucosinolates were identified from the extract(the total glucosinolates content was 73.7%)including 2-allyl glucosinolate,4-methyl thiobutyl glucosinolate and 4-methoxy 3-indolemethyl glucosinolate.The glucosinolate extract had no inhibitory effect on normal human colonic myofibroblasts CCD-18Co cells,but led to cell cycle arrest in HCT116 cells by down-regulating cell cycle genes and related signaling proteins(cyclin B,cyclinD1,and led to the apoptosis of HCT116 cells cyclinE)by up-regulating apoptotic genes and related signaling proteins(caspase-3,and cleaved caspase-3),thereby inhibiting cell proliferation.

关 键 词：高菜 硫代葡萄糖苷 人结肠癌细胞株HCT116 细胞凋亡 增殖 结肠癌 

分 类 号：R735.35[医药卫生—肿瘤]