新的蒽醌类衍生物1-硝基-2-酰基蒽醌-苯丙氨酸通过抑制NF-κB/p65信号通路降低乳腺癌MCF-7细胞的迁移能力  

A Novel Anthraquinone Derivative 1-Nitro-2-AcylanthraquinonePhenylalanine Inhibits Human Breast Cancer MCF-7 Cell Migration by Blocking NF-κB/p65 Pathway

作　　者：李玉英[1] 牛敏[1] 张立伟[1] 王转花[1] 

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

出　　处：《中国生物化学与分子生物学报》2017年第3期269-277,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.31171659,No.31600631);山西省自然科学基金项目(No.201601D011067,No.2015021047)资助~~

摘　　要：肿瘤细胞的侵袭和转移与大多数癌症患者的死亡率密切相关。了解肿瘤细胞的迁移机制可为阻断肿瘤细胞的转移提供一个关键的策略。蒽醌衍生物具有一定的抗肿瘤作用,我们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酰胺蒽醌衍生物1-硝基-2-酰基蒽醌-苯丙氨酸(简称C7),发现其具有很好的抗肿瘤活性。为了探究蒽醌类衍生物C7对人乳腺癌MCF-7细胞迁移的作用及其机制,本文首先采用MTT比色法检测蒽醌类衍生物C7对人乳腺癌细胞MCF-7生长活力的影响,结果证明较高浓度(60~100μg/m L)的蒽醌类衍生物C7对乳腺癌MCF-7细胞的增殖具有明显的抑制作用。其次,采用细胞划痕实验检测C7对MCF-7细胞迁移的影响,发现较低浓度(20~40μg/m L)的蒽醌类衍生物C7可以显著降低MCF-7细胞的迁移率。为进一步探究C7抑制MCF-7细胞迁移的分子机制,通过免疫荧光技术检测NF-κB/p65蛋白的核转位情况;同时利用qRT-PCR及Western印迹实验检测C7对MCF-7细胞中NF-κB/p65通路及迁移相关基因和蛋白质表达的影响。结果表明C7可以下调细胞质中IκBα的磷酸化,降低NF-κB/p65蛋白的核转位,减小MMP-2和MMP-9蛋白的表达。因此,C7可能是通过抑制NF-κB/p65信号通路的活化,抑制MMP-2和MMP-9蛋白的表达,进而抑制MCF-7细胞的迁移。Metastasis and invasion of cancer cells are closely related to the cancer mortality.Understanding the migration mechanism of cancer cells could provide a key treatment strategy for the cancer cells metastasis. Anthraquinone derivatives have certain anti-cancer effect. According to the mechanism of anti-cancer drugs and the structure-activity relationship of anthraquinone derivatives,an anthraquinone derivative 1-nitro-2-acylanthraquinone-phenylalanine named C7 has been designed and synthesized. We investigated the migration mechanism of anthraquinone derivative C7 in human breast cancer cell. First,the effects of C7 on the viability of MCF-7 cells were tested by MTT assay. The resultshowed that the high concentration（ 60- 100 μg / m L） of C7 could significantly inhibit the viability of MCF-7 cells. Second,the wound-healing assay was used to determine cell migration. It showed that the low concentration（ 20- 40 μg / m L） of C7 decreased the migration rate of MCF-7 cells. Furthermore,an immunofluorescence assay was performed via an anti-p65 monoclonal antibody,and we detected the expression of genes and proteins of migration and NF-κB / p65 pathway by qRT-PCR and Western blotting. As the result,anthraquinone derivative C7 suppressed the nuclear translocation of NF-κB / p65 by inhibiting the phosphorylation of IκBα,downregulated the expression of matrix metalloproteinase（ MMP）-2 /9 at gene and protein level. Taken together,these data indicated that anthraquinone derivative C7 could block NF-κB / p65 signal pathway,thereby inhibiting MCF-7 cells migration.

关 键 词：蒽醌类衍生物 迁移 基质金属蛋白酶2 基质金属蛋白酶9 NF-ΚB/P65 

分 类 号：Q786[生物学—分子生物学]