异硫氰酸异丁酯抑制肝癌细胞HepG2生长的效应与机理  

Effect and mechanism of isobutyl isothiocyanate on inhibiting the growth of HepG2

作　　者：周巧巧[1,2] 匡政坤 张洪权 熊丽[3] 周诗毅 ZHOU Qiaoqiao;KUANG Zhengkun;ZHANG Hongquan;XIONG Li;ZHOU Shiyi(School of Chemistry and Life Sciences,Hubei University of Education,Wuhan 430205,China;Hubei Key Laboratory of Purification and Application of Plant Anticancer Active Ingredients,School of Chemistry and Life Sciences,Hubei University of Education,Wuhan 430205,China;College of Life Science,Central China Normal University,Wuhan 430079,China)

机构地区：[1]湖北第二师范学院化学与生命科学学院,武汉430205 [2]湖北第二师范学院化学与生命科学学院植物抗癌活性物质提纯与应用湖北省重点实验室,武汉430205 [3]华中师范大学生命科学学院,武汉430079

出　　处：《华中师范大学学报（自然科学版）》2022年第5期834-840,共7页Journal of Central China Normal University：Natural Sciences

基　　金：湖北省教育厅科学研究计划项目(B2019183).

摘　　要：为了探究异硫氰酸异丁酯(isobutyl isothiocyanate)对肝癌细胞HepG2的生长抑制效应及其分子机制,该研究通过MTT实验检测化合物对细胞的生长抑制率,计算出IC_(50)值为3.5μg·mL^(-1);通过流式细胞技术检测发现0.437μg·mL^(-1)以上浓度的化合物能诱导HepG2细胞凋亡;通过划痕实验检测发现0.875μg·mL^(-1)以上浓度的化合物能抑制细胞迁移;进一步通过生物信息学分析表明异硫氰酸异丁酯的靶蛋白可能为巨噬细胞迁移抑制因子(MIF),且HepG2细胞中MIF表达量高于低敏感株.免疫印迹实验也进一步发现化合物不仅能够抑制蛋白酪氨酸激酶2/信号转导子和转录激活子3(JAK2/STAT3)信号通路的激活,还能下调p53蛋白表达.研究结果表明,异硫氰酸异丁酯可能通过靶向HepG2细胞中的MIF蛋白,影响MIF与其受体CD74相互作用,从而下调p53蛋白表达,阻滞细胞周期的正常进行,诱导细胞凋亡,对肝癌具有特异性的抑癌潜力.In order to explore the inhibitory effect of isobutyl isothiocyanate on the growth of primary hepatocellular carcinoma cell line HepG2 and related molecular mechanism,the growth inhibition rate of the compound on the cells was detected by the MTT assays with the IC_(50) value of 3.5μg·mL^(-1).Through flow cytometry assay,it was found that isobutyl isothiocyanate induced cellular apoptosis at concentrations above 0.437μg·mL^(-1).Moreover,scratch test showed that isobutyl isothiocyanate inhibited cell migration at concentrations above 0.875μg·mL^(-1).Further bioinformatics analysis showed that the target protein of this compound might be macrophage migration inhibitory factor(MIF).And the expression level of MIF in HepG2 cells was higher than that in low-susceptibility strains.Western blot assays suggested that isobutyl isothiocyanate was able to not only inhibit Janus kinases/just another kinases 2/signal transducers and activators of transcription 3(JAK2/STAT3)signaling,but also down-regulate the expression of p53 protein.These data indicated that isobutyl isothiocyanate might target the MIF of HepG2 cells to inhibit the interaction between MIF and CD74,down-regulating p53,thereby blocking the normal cell cycle and inducing cell apoptosis,suggesting it to be a potential suppressor to liver cancer.

关 键 词：异硫氰酸异丁酯 HEPG2细胞 JAK2/STAT3信号通路 MIF蛋白 细胞周期 细胞凋亡 

分 类 号：Q291[生物学—细胞生物学]