Benzimidazoles Downregulate Mdm2 and MdmX and Activate p53 in MdmX Overexpressing Tumor Cells
Benzimidazoles Downregulate Mdm2 and MdmX and Activate p53 in MdmX Overexpressing Tumor Cells
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DOI：
10.3390/molecules24112152
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发表时间：
2019-06-01
期刊：
影响因子：
4.6
通讯作者：
Slaninova, Iva
中科院分区：
文献类型：
--
作者：
Mrkvova, Zuzana;Uldrijan, Stjepan;Slaninova, Iva
肿瘤抑制基因p53在约50%的癌症中发生突变。大多数恶性黑色素瘤携带野生型p53，但p53活性通常由于其负调节因子Mdm 2或MdmX的过表达而受到抑制。我们在携带p53活性荧光素酶报告基因构建体的A375细胞上进行了2448种化合物的高通量筛选，以揭示促进黑素瘤中p53活性的化合物。阿苯达唑和芬苯达唑，两个批准的和常用的苯并咪唑驱虫剂，刺激p53活性，并选择进行进一步的研究。阿苯达唑和芬苯达唑治疗后p53和p21蛋白水平增加，表明p53-p21通路的激活，而在过表达这些蛋白的黑色素瘤和乳腺癌细胞中Mdm 2和MdmX的水平降低。我们还观察到细胞活力的降低和细胞形态的变化，对应于有丝分裂灾难，即，G2/M期细胞周期阻滞的大型多核细胞与破坏微管。总之，我们建立了一个新的工具来测试小分子化合物对p53活性的影响，并使用它来确定苯并咪唑类药物在黑色素瘤细胞中的作用。这些药物通过下调过表达这些蛋白质的细胞中的负调节因子Mdm 2和MdmX来促进野生型p53的稳定性和转录活性。结果表明，苯并咪唑驱肠虫剂用于治疗过表达p53负调节因子的癌症的潜力。
Tumor suppressor p53 is mutated in about 50% of cancers. Most malignant melanomas carry wild-type p53, but p53 activity is often inhibited due to overexpression of its negative regulators Mdm2 or MdmX. We performed high throughput screening of 2448 compounds on A375 cells carrying p53 activity luciferase reporter construct to reveal compounds that promote p53 activity in melanoma. Albendazole and fenbendazole, two approved and commonly used benzimidazole anthelmintics, stimulated p53 activity and were selected for further studies. The protein levels of p53 and p21 increased upon the treatment with albendazole and fenbendazole, indicating activation of the p53-p21 pathway, while the levels of Mdm2 and MdmX decreased in melanoma and breast cancer cells overexpressing these proteins. We also observed a reduction of cell viability and changes of cellular morphology corresponding to mitotic catastrophe, i.e., G2/M cell cycle arrest of large multinucleated cells with disrupted microtubules. In summary, we established a new tool for testing the impact of small molecule compounds on the activity of p53 and used it to identify the action of benzimidazoles in melanoma cells. The drugs promoted the stability and transcriptional activity of wild-type p53 via downregulation of its negative regulators Mdm2 and MdmX in cells overexpressing these proteins. The results indicate the potential for repurposing the benzimidazole anthelmintics for the treatment of cancers overexpressing p53 negative regulators.