Department of Biomedical Science, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India.
Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, Republic of Korea.
Eur J Pharm Sci. 2018 Apr 30;116:2-14. doi: 10.1016/j.ejps.2017.09.019. Epub 2017 Sep 12.
In spite of advances in breast cancer treatment and early diagnosis, drug toxicity, cancer relapse, multidrug resistance and metastasis are the major impediment to the developments of efficient drugs. However, unique druggable targets of cancer cells distinct from the normal cells provide new rationale in cancer treatment. Previous reports clearly emphasize the differential expression and localization of Y box binding protein-1 (YB-1) between normal breast tissues and different stages of breast cancer. Y box binding protein-1 is DNA as well as RNA binding protein involved in transcription and translation regulation of various proteins involved in cancer progression, apoptosis, cell cycle, epithelial to mesenchymal transition (EMT) and drug resistance. Particularly, during doxorubicin (DOX) treatment and cancer relapse conditions, YB-1 expression was very high in breast cancer tissues and localized in to nucleus which further favours DOX efflux and metastasis. Moreover, siRNA mediated silencing of YB-1 reduces breast cancer progression and metastasis. In this rationale, using an array of computational methods, 2,4-dihydroxy-5-pyrimidinyl imidothiocarbomate (DPI) has been screened out as a drug-likeness antagonist to the YB-1for cancer treatment. In this study, we determined that DPI was toxic to breast cancer cell lines as individual drug as well as in combination with DOX. Moreover, immunofluorescence and confocal studies showed that DPI decreases DOX induced YB-1 nuclear translocation and increases DOX accumulation in breast cancer cell line. A G1/G0 phase cell cycle arrest and apoptosis was also induced by DPI. Moreover, DPI modulated YB-1 downstream targets such as p53, caspase-3, CDK-1 which are involved in cell cycle progression and apoptosis. Further, metastatic functional analysis revealed that DPI inhibits cell adhesion, migration, invasion in aggressive metastatic cell line and inhibits angiogenesis in chick embryonic chorioallantoic membrane (CAM) model. Meanwhile, DPI alters the expression of YB-1 downstream targets which are involved in metastasis such as VEGFR, caveolin, E-cadherin, cytokeratins, desmin and vimentin in MDA-MB-231 xenograft in chick embryonic CAM membrane. The results clearly demonstrated that DPI inhibited YB-1 nuclear translocation, thereby exhibited anti-apoptotic, anti-proliferative and anti-metastatic activities and increases the therapeutic potential of commercial breast cancer drug doxorubicin.
尽管乳腺癌治疗和早期诊断取得了进展，但药物毒性、癌症复发、多药耐药和转移仍是高效药物开发的主要障碍。然而，癌细胞特有的、不同于正常细胞的可用药靶为癌症治疗提供了新的依据。先前的报告清楚地强调了 Y 盒结合蛋白-1（YB-1）在正常乳腺组织和不同阶段乳腺癌之间的差异表达和定位。YB-1 是一种 DNA 和 RNA 结合蛋白，参与涉及癌症进展、细胞凋亡、细胞周期、上皮间质转化（EMT）和耐药性的各种蛋白质的转录和翻译调控。特别是在阿霉素（DOX）治疗和癌症复发期间，乳腺癌组织中 YB-1 的表达非常高，并且定位于核内，这进一步有利于 DOX 外排和转移。此外，YB-1 的 siRNA 介导沉默可降低乳腺癌的进展和转移。基于此，本研究使用一系列计算方法筛选出 2,4-二羟基-5-嘧啶基亚氨基硫代甲酰胺（DPI）作为 YB-1 的药物样拮抗剂用于癌症治疗。在这项研究中，我们确定 DPI 作为单一药物以及与 DOX 联合使用对乳腺癌细胞系均具有毒性。此外，免疫荧光和共聚焦研究表明，DPI 可降低 DOX 诱导的 YB-1 核转位并增加乳腺癌细胞系中 DOX 的积累。DPI 还诱导 G1/G0 期细胞周期停滞和细胞凋亡。此外，DPI 调节 YB-1 下游靶标，如参与细胞周期进程和凋亡的 p53、caspase-3、CDK-1。进一步的转移功能分析表明，DPI 抑制侵袭性转移细胞系的细胞黏附、迁移和侵袭，并抑制鸡胚绒毛尿囊膜（CAM）模型中的血管生成。同时，DPI 改变了 YB-1 下游参与转移的靶标，如在鸡胚 CAM 膜中 MDA-MB-231 异种移植物中的 VEGFR、 caveolin、E-cadherin、细胞角蛋白、结蛋白和波形蛋白的表达。结果清楚地表明，DPI 抑制 YB-1 核转位，从而表现出抗凋亡、抗增殖和抗转移活性，并增加了商业乳腺癌药物阿霉素的治疗潜力。
