Casein Kinase-1-Alpha Inhibitor (D4476) Sensitizes Microsatellite Instable Colorectal Cancer Cells to 5-Fluorouracil via Authophagy Flux Inhibition

IF 2.9 4区医学 Q3 IMMUNOLOGY

Archivum Immunologiae et Therapiae Experimentalis Pub Date : 2021-09-18 DOI:10.1007/s00005-021-00629-2

Morvarid Siri, Hamid Behrouj, Sanaz Dastghaib, Mozhdeh Zamani, Wirginia Likus, Sedigheh Rezaie, Jacek Hudecki, Saeed Khazayel, Marek J. Łos, Pooneh Mokarram, Saeid Ghavami

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引用次数: 16

Abstract

Adjuvant chemotherapy with 5-fluorouracil (5-FU) does not improve survival of patients suffering from a form of colorectal cancer (CRC) characterized by high level of microsatellite instability (MSI-H). Given the importance of autophagy and multi-drug-resistant (MDR) proteins in chemotherapy resistance, as well as the role of casein kinase 1-alpha (CK1α) in the regulation of autophagy, we tested the combined effect of 5-FU and CK1α inhibitor (D4476) on HCT116 cells as a model of MSI-H colorectal cancer. To achieve this goal, the gene expression of Beclin1 and MDR genes, ABCG2 and ABCC3 were analyzed using quantitative real-time polymerase chain reaction. We used immunoblotting to measure autophagy flux (LC3, p62) and flow cytometry to detect apoptosis. Our findings showed that combination treatment with 5-FU and D4476 inhibited autophagy flux. Moreover, 5-FU and D4476 combination therapy induced G2, S and G1 phase arrests and it depleted mRNA of both cell proliferation-related genes and MDR-related genes (ABCG2, cyclin D1 and c-myc). Hence, our data indicates that targeting of CK1α may increase the sensitivity of HCT116 cells to 5-FU. To our knowledge, this is the first description of sensitization of CRC cells to 5-FU chemotherapy by CK1α inhibitor.

Graphic abstract

Abstract Image

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酪蛋白激酶-1- α抑制剂(D4476)通过自噬通量抑制微卫星不稳定结直肠癌细胞对5-氟尿嘧啶的敏化

5-氟尿嘧啶（5-FU）辅助化疗不能提高患有以高水平微卫星不稳定性（MSI-H）为特征的结直肠癌（CRC）的患者的生存率。鉴于自噬和多药耐药（MDR）蛋白在化疗耐药性中的重要性，以及酪蛋白激酶1-α（CK1α）在自噬调节中的作用，我们测试了5-FU和CK1α抑制剂（D4476）对作为MSI-H结直肠癌癌症模型的HCT116细胞的联合作用。为了实现这一目标，使用定量实时聚合酶链反应分析Beclin1和MDR基因ABCG2和ABCC3的基因表达。我们使用免疫印迹法测量自噬流量（LC3，p62），并使用流式细胞术检测细胞凋亡。我们的研究结果表明，5-FU和D4476联合治疗可抑制自噬流量。此外，5-FU和D4476联合治疗诱导了G2、S和G1期阻滞，并耗尽了细胞增殖相关基因和MDR相关基因（ABCG2、细胞周期蛋白D1和c-myc）的mRNA。因此，我们的数据表明，靶向CK1α可能增加HCT116细胞对5-FU的敏感性。据我们所知，这是首次描述CK1α抑制剂使CRC细胞对5-FU化疗的敏感性。图形摘要

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来源期刊

Archivum Immunologiae et Therapiae Experimentalis 医学-免疫学

CiteScore

5.90

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Archivum Immunologiae et Therapiae Experimentalis (AITE), founded in 1953 by Ludwik Hirszfeld, is a bimonthly, multidisciplinary journal. It publishes reviews and full original papers dealing with immunology, experimental therapy, immunogenetics, transplantation, microbiology, immunochemistry and ethics in science.