Novel small molecule decreases cell proliferation, migration, clone formation, and gene expression through ERK inhibition in MCF-7 and MDA-MB-231 breast cancer cell lines
Novel small molecule decreases cell proliferation, migration, clone formation, and gene expression through ERK inhibition in MCF-7 and MDA-MB-231 breast cancer cell lines
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DOI：
10.1097/cad.0000000000000766
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发表时间：
2019-07-01
影响因子：
2.3
通讯作者：
Papasotiriou, Ioannis
中科院分区：
文献类型：
--
作者：
Hatzidaki, Eleana;Parsonidis, Panagiotis;Papasotiriou, Ioannis
Ras-Raf-MEK1/2-ERK1/2通路在人类肿瘤中广泛存在，是抗癌药物开发的重要靶点。然而，尽管最初的临床反应，很可能是由于串扰通路的激活，但往往会产生耐药性。在研究遗传癌症中心(RGCC)，我们正在合成一种新的ERK抑制剂，针对该途径的最后阶段，从而将串扰降至最低。我们合成了中间体分子RGCC416，并对其生物活性进行了测试。采用MCF-7和MDA-MB-231细胞。结晶紫法测定细胞存活率，四甲基偶氮唑蓝比色法测定不同浓度化合物对细胞增殖的影响。细胞迁移和集落形成分别评估癌细胞的侵袭能力和单个癌细胞的生长能力。用聚合酶链式反应检测MAPK/PI3K通路相关基因的表达。免疫印迹法检测ERK和磷酸化ERK在胞浆和胞核的表达。结果发现，尽管该化合物不影响细胞活力，但它显著抑制了两种细胞系的细胞增殖、迁移和集落形成。在MDA-MB-231中，这可能是通过将磷酸化ERK保留在细胞质中，在那里它不能激活与癌症相关的基因。MCF-7细胞ERK表达水平无明显差异。这可能是因为这些细胞用于生存的不同途径。我们已经合成了一种分子，它可能是一种很有前途的ERK抑制剂，从而为乳腺癌患者带来可能的新治疗选择。
The Ras-Raf-MEK1/2-ERK1/2 pathway is an attractive target for the development of anticancer agents because of the high prevalence of ERK activation in human cancers. However, resistance is often developed despite initial clinical response, most likely because of activation of cross-talk pathways. In Research Genetic Cancer Center (RGCC), we are in the process of synthesizing a novel ERK inhibitor, targeting the final stage of the pathway, thus minimizing cross-talk. We have synthesized an intermediate molecule -RGCC416 - and tested its biological activity. MCF-7 and MDA-MB-231 cells were used. Cell viability was measured by crystal violet and cell proliferation by the methyl tetrazolium assay using various compound concentrations. Cell migration and colony formation were determined to assess the ability of invasion and single cancer cell growth, respectively. Expression of genes linked to MAPK/PI3K pathways was determined by PCR. ERK and phospho-ERK levels were determined in both the cytoplasm and the nucleus by western blot. It was found that although the compound did not affect viability, it significantly decreased cell proliferation, migration, and colony formation in both cell lines. In MDA-MB-231, this is possibly through retaining phospho-ERK to the cytoplasm, where it cannot activate cancer-associated genes. There was no difference in ERK levels in MCF-7 cells. This could be because of the different pathways that these cells utilize for survival. We have synthesized a molecule, which could be a promising ERK inhibitor, leading to possible novel treatment options for breast cancer patients.