JIB-04通过调控细胞周期蛋白抑制结肠癌细胞增殖  

JIB-04 Inhibits the Proliferation of Colon Cancer Cells by Regulating Cell Cycle Proteins

作　　者：叶春雨 朴光宇 金艳花 YE Chun-Yu;PIAO Guang-Yu;JIN Yan-Yua(Department of Cell Biology and Medical Genetics,College of Medicine,Yanbian University,Yanji 133002,Jilin,China)

机构地区：[1]延边大学医学院细胞生物学与医学遗传学教研室,吉林延吉133002

出　　处：《中国生物化学与分子生物学报》2020年第7期795-803,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.81660494)资助。

摘　　要：已有报道显示,组蛋白去甲基化酶抑制剂JIB-04(Jumonji histone demethylase inihibitor,JIB-04)抑制肿瘤的发生发展,但其具体作用机制仍不清楚。本研究以结肠癌细胞HCT116和HT29为对象,探讨组蛋白去甲基化酶抑制剂JIB-04对结肠癌细胞增殖的影响,并阐述其作用机制。MTT和平板克隆形成实验显示,JIB-04以浓度和时间依赖的方式降低HCT116和HT29细胞的增殖能力,半数抑制浓度分别为661.7 nmol/L及226.1 nmol/L,细胞克隆数也明显减少。qRT-PCR和Western印迹结果证明,与未经JIB-04处理的HCT116和HT29细胞比较,细胞周期蛋白D1(cyclinD1)、细胞周期蛋白E1(cyclinE1)的mRNA水平和CDK4、CDK6及细胞周期蛋白D1的蛋白质水平有不同程度的降低,同时CDK抑制剂p21的蛋白质水平分别上调约1.99倍和2.37倍。检测凋亡相关因子发现,在HCT116和HT29细胞中,p53、胱天蛋白酶3、胱天蛋白酶9、Bax、JNK及PUMA的mRNA及蛋白质水平均降低。机制研究显示,JIB-04使组蛋白赖氨酸特异性去甲基化酶1(LSD1)的mRNA水平分别下调约56.8%和75.5%,其蛋白质水平下调约27.12%和15.32%。本研究结果表明,JIB-04可改变组蛋白赖氨酸特异性去甲基化酶LSD1的活性,调控细胞周期蛋白及相关因子在mRNA和蛋白质水平的表达,同时诱导凋亡蛋白质发生改变,从而抑制结肠癌细胞增殖和凋亡。It has been reported that Jumonji histone demethylase inihibitor(JIB-04)inhibits the occurrence and development of tumors,but the specific mechanism is still unclear.In this paper,colon cancer cells HCT116 and HT29 were used as the models,and the effect of JIB-04 on the proliferation of colon cancer cells was explored and its mechanism was explained.MTT assays and plate cloning formation experiments showed that JIB-04 reduced the proliferation ability of HCT116 and HT29 cells in a concentration-and time-dependent manner,with the half-inhibitory concentrations being 661.7 nmol/L and 226.1 nmol/L,respectively.And the number of cell clones also decreased significantly.qRT-PCR and Western blotting results showed that compared with HCT116 and HT29 cells not treated with JIB-04,the mRNA levels of the cyclinD1,cyclinE1,and CDK4,CDK6,and cyclinD1 proteins levels were reduced to different degrees.At the same time,p21(CDK inhibitor)expression increased by about 1.99-fold and 2.37-fold,respectively.Detection of apoptosis-related factors revealed that the mRNA and protein levels of p53,caspase-3,caspase-9,Bax,JNK and PUMA were reduced in HCT116 and HT29 cells.Mechanistic analysis found that JIB-04 reduced histone demethylase LSD1 mRNA levels by about 56.8%and 75.5%,and its protein levels were reduced by about 27.12%and 15.32%.The results of this study indicate that JIB-04 can alter the activity of LSD1 and regulate mRNA and protein levels of cyclins and related factors,and it induces changes in apoptotic proteins,thereby inhibiting colon cancer cell proliferation and apoptosis.

关 键 词：组蛋白去甲基化酶抑制剂 结肠癌 赖氨酸特异性去甲基酶1 细胞周期 

分 类 号：R34[医药卫生—基础医学]