Fluvastatin exerts an antitumor effect in vemurafenib-resistant melanoma cells
Fluvastatin exerts an antitumor effect in vemurafenib-resistant melanoma cells
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DOI：
10.1097/cad.0000000000000757
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发表时间：
2019-06-01
影响因子：
2.3
通讯作者：
Maesawa, Chihaya
中科院分区：
文献类型：
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作者：
Nishiya, Masao;Yasuhira, Shinji;Maesawa, Chihaya
尽管vemurafenib已被证明可以提高携带BRAF V600E突变的转移性黑色素瘤患者的总体生存率，但其疗效往往受到通过几种不同机制在相对较短时间内获得的耐药性的阻碍。在本研究中，我们利用培养细胞系模型研究了氟伐他汀作为克服这种获得性耐药性的可能策略的作用。我们从三个BRAF （V600E）突变的黑色素瘤细胞系（C32、HMY-1和SK-MEL-28）中建立了vemurafenib耐药（VR）细胞，并通过水溶性四氮唑盐试验、western blot、实时定量PCR和免疫荧光显微镜评估了VR细胞获得性耐药的机制。通过计算联合指数和western blot评价vemurafenib和氟伐他汀联合使用对亲本细胞和VR细胞的生长抑制作用。3个VR细胞的IC50值均比亲本细胞高5 ~ 100倍。HMY-1和SK-MEL-28衍生的VR细胞表现出AKT激酶的组成性激活，特异性AKT抑制剂MK-2208或PI3K抑制剂wortmannin增加了细胞对vemurafenib的敏感性。有趣的是，应用他汀类相关药物氟伐他汀也可能通过减轻构成型AKT激活而导致VR细胞对vemurafenib敏感性的协同增加（联合指数：0.73-0.86），而同样的治疗并没有显著改变亲代细胞对vemurafenib的敏感性。我们的研究结果表明，他汀类药物可能有助于克服通过PI3K-AKT轴的组成性激活获得的vemurafenib耐药性。版权所有(c) 2019威科集团健康有限公司版权所有。
Although vemurafenib has been shown to improve the overall survival of patients with metastatic melanoma harboring the BRAF V600E mutation, its efficacy is often hampered by drug resistance acquired within a relatively short period through several distinct mechanisms. In the present study, we investigated the effect of fluvastatin as a possible strategy to overcome such acquired resistance using a cultured cell line model. We established vemurafenib-resistant (VR) cells from three BRAF (V600E)-mutated melanoma lines (C32, HMY-1, and SK-MEL-28) and evaluated the mechanism of acquired resistance of VR cells by water-soluble tetrazolium salts assay, western blot, realtime quantitative PCR, and immunofluorescent microscopy. The efficacy of the combination of growth inhibitory effect of vemurafenib and fluvastatin on respective parental and VR cells were assessed by calculating combination index and western blot. IC50 values of three VR cells were 5-100-fold higher than those for the respective parental cells. The VR cells derived from HMY-1 and SK-MEL-28 showed constitutive activation of AKT kinase, and the specific AKT inhibitor MK-2208 or the PI3K inhibitor wortmannin increased the cellular sensitivity to vemurafenib. Intriguingly, application of a statin-related drug, fluvastatin, also resulted in a synergistic increase of sensitivity to vemurafenib in the VR cells (combination index: 0.73-0.86) probably by alleviating constitutive AKT activation, whereas the same treatment did not notably alter the vemurafenib sensitivity of the parental cells. Our results suggest the possible usefulness of statin-related drugs for overcoming vemurafenib resistance acquired through constitutive activation of the PI3K-AKT axis. Copyright (c) 2019 Wolters Kluwer Health, Inc. All rights reserved.