1-硝基-2-酰基蒽醌-缬氨酸通过抑制ERK1/2激活和ERCC1表达诱导结肠癌细胞死亡  

1-Nitro-2-Acylanthraquinone-Valine Induces Human Colon Cancer Cell Deaths via the ERK1/2-ERCC1 Signaling Pathway

作　　者：李玉英 郑婕 张立伟[2] 王转花 LI Yu-Ying;ZHENG Jie;ZHANG Li-Wei;WANG Zhuan-Hua(Key Laboratory of Chemical Biology and Molecular Engineering,Ministry of Education,Institute of Biotechnology, Shanxi University,Taiyuan 030006,China;Institute of Molecular Science ,Shanxi University ,Taiyuan 030006,China)

机构地区：[1]化学生物学与分子工程教育部重点实验室山西大学生物技术研究所,太原030006 [2]山西大学分子科学研究所,太原030006

出　　处：《中国生物化学与分子生物学报》2019年第1期81-91,共11页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.31600631);山西省自然科学基金项目(No.201601D011067)资助~~

摘　　要：化学方法合成是新药研发的一种重要途径。结合抗肿瘤药物的作用机制以及蒽醌类衍生物的构效关系,设计合成了一类新的蒽醌类衍生物1-硝基-2-酰基蒽醌-缬氨酸(简称C3),发现其具有很好的抗肿瘤活性。为了确定蒽醌类衍生物C3对结肠癌HCT116和HT29细胞的作用及其分子机制,首先通过MTT比色法检测C3对结肠癌HCT116和HT29细胞活性的影响。结果显示,C3对这两种结肠癌细胞具有明显的抑制作用,呈时间和剂量依赖性。60μg/mL的C3处理HCT116和HT29细胞48 h,细胞活性分别是50.67%和59.77%,达到了半抑制浓度;同时,其细胞形态和细胞核发生明显变化。进一步采用Western印迹和qRT-PCR技术,检测C3对DNA切除修复交叉互补1(excision repair cross-complementation group 1,ERCC1)转录水平和蛋白质水平表达及其稳定性的影响。结果表明,C3降低了ERCC1转录水平和蛋白质水平的表达,并且减弱了ERCC1转录水平和蛋白质水平的稳定性。最后,用U0126(MEK1/2抑制剂)和C3联合作用结肠癌HCT116和HT29细胞,通过Western印迹检测ERCC1蛋白质水平的表达。结果表明,C3通过降低p-ERK1/2的蛋白质水平的表达,从而抑制ERCC1的表达。上述结果证明,C3通过细胞外调节蛋白激酶(extracellular regulated protein kinases, ERK1/2)信号通路,降低了ERCC1转录水平和蛋白质水平的稳定性,使ERCC1转录水平和蛋白质水平表达发生下调,进而抑制结肠癌HCT116和HT29细胞的活性。Chemical synthesis is an important way to develop new drugs. A new class of anthraquinone derivatives, 1-nitro-2-acylanthraquinone-valine(C3), was designed and synthesized by combining the mechanism of antitumor drugs and the structure-activity relationship of anthraquinone derivatives. It was found that C3 has good antitumor activity. Therefore, we aim to determine the effect of anthraquinone derivative C3 on human colon cancer HCT116 and HT29 cells and its molecular mechanism. Firstly, the effects of C3 on the activity of colon cancer HCT116 and HT29 cells were detected by MTT colorimetry. The results showed that C3 could inhibit these two kinds of colon cancer cells in a time-and dose-dependent manner. Moreover, HCT116 and HT29 cells were treated with C3 at 60 μg/mL for 48 hours, and the cell viability was 50.67% and 59.77% respectively, which reached a semi-inhibitory concentration. At the same time, cellular and nuclear morphology changed significantly. Western blotting and qRT-PCR were used to detect the effects of C3 on the expression of ERCC1(excision repair cross-complementation group 1) and stability of ERCC1 protein levels. The results showed that both the transcriptional and protein levels were decreased. Finally, HCT116 and HT29 cells were treated with U0126(MEK1/2 inhibitor) and C3. The abundance of ERCC1 proteins was detected by Western blotting. The results showed that C3 inhibited the expression of ERCC1 by reducing p-ERK1/2 proteins. These results suggest that C3 reduces ERCC1 transcription levels and ERCC1 protein levels through the ERK1/2(extracellular regulated protein kinase) signaling pathway, and down-regulates the expression of ERCC1 and its protein levels, thereby inhibiting the activity of HCT116 and HT29 cells.

关 键 词：蒽醌类衍生物 切除修复交叉互补1 细胞外调节蛋白激酶 结肠癌 

分 类 号：Q74[生物学—分子生物学]