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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。
Tumor cell invasion and metastasis are closely related to the mortality of most cancer patients. Understanding the mechanism of tumor cell migration can provide a key strategy for blocking the metastasis of tumor cells. Anthraquinone derivatives have certain anti-tumor effect, we combined with the mechanism of antitumor drugs and the structure-activity relationship of anthraquinone derivatives, designed and synthesized a new class of amide anthraquinone derivatives 1-nitro-2-acyl Anthraquinone-phenylalanine (referred to as C7), found that it has good antitumor activity. In order to explore the effect of anthraquinone derivative C7 on human breast cancer MCF-7 cell migration and its mechanism, this article first using MTT colorimetric assay anthraquinone derivative C7 on human breast cancer cell MCF-7 growth activity, The results showed that higher concentration (60 ~ 100μg / m L) anthraquinone derivatives C7 breast cancer MCF-7 cells significantly inhibited the proliferation. Secondly, the effect of C7 on the migration of MCF-7 cells was detected by cell scratch assay. It was found that C7, a derivative of anthraquinone at a lower concentration (20-40 μg / mL), could significantly reduce the MCF-7 cell migration. To further explore the molecular mechanism of C7 inhibiting MCF-7 cell migration, the nuclear translocation of NF-κB / p65 protein was detected by immunofluorescence technique. Meanwhile, the effect of C7 on NF-κB / p65 protein expression was detected by qRT-PCR and Western blotting. κB / p65 pathway and migration related genes and protein expression. The results showed that C7 down-regulated the phosphorylation of IκBα in the cytoplasm, decreased the nuclear translocation of NF-κB / p65 protein and decreased the expression of MMP-2 and MMP-9. Therefore, C7 may inhibit the migration of MCF-7 cells by inhibiting the activation of NF-κB / p65 signaling pathway and inhibiting the expression of MMP-2 and MMP-9 proteins.