Siva-1 regulates multidrug resistance of gastric cancer by targeting MDR1 and MRP1 via the NF-κB pathway
Siva-1 regulates multidrug resistance of gastric cancer by targeting MDR1 and MRP1 via the NF-κB pathway
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DOI：
10.3892/mmr.2020.11211
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发表时间：
2020-08-01
影响因子：
3.4
通讯作者：
Mai, Wei
中科院分区：
文献类型：
--
作者：
Kong, Fan-Biao;Deng, Qiao-Ming;Mai, Wei
Siva-1是一种众所周知的抗凋亡蛋白，在多种类型的癌细胞中发挥作用。然而，Siva-1是否通过NF-κ B通路影响胃癌的多药耐药目前尚不清楚。本研究旨在探讨Siva-1在体外胃癌耐药中的作用。建立了稳定过表达Siva-1的长春新碱（VCR）耐药KATO III/VCR胃癌细胞系。Western blotting检测Siva-1、NF-κ B B、多药耐药蛋白1（MDR 1）和多药耐药蛋白1（MRP 1）的蛋白表达水平。通过测量KATO III/VCR细胞对VCR、5-氟尿嘧啶和阿霉素的50%抑制浓度，评估Siva-1过表达对抗癌药物耐药性的影响。流式细胞仪检测阿霉素外排率和细胞凋亡率。采用集落形成法、创伤愈合法和Transwell法分别检测细胞的增殖、迁移和侵袭能力。本研究结果显示，Siva-1过表达的KATO III/VCR胃癌细胞对VCR、5-氟尿嘧啶和阿霉素的敏感性显著降低。流式细胞仪检测结果显示，Siva-1过表达后，凋亡细胞的百分比下降。殖民地形成实验表明，Siva-1过表达能显著促进细胞生长和增殖。此外，Siva-1过表达增加了KATO III/VCR细胞的体外迁移和侵袭。Western印迹分析确定Siva-1过表达增加NF-κ B B、MDR 1和MRP 1水平。目前的研究表明，Siva-1的过表达，作为胃癌细胞中的MDR 1和MRP 1基因表达的调节剂，通过促进NF-κ B表达，抑制胃癌细胞对某些化疗药物的敏感性。这些结果为胃癌发生的分子机制提供了新的认识，对胃癌的临床诊断和治疗具有重要意义。
Siva-1 is a well-known anti-apoptosis protein that serves a role in multiple types of cancer cells. However, whether Siva-1 affects multidrug resistance via the NF-kappa B pathway in gastric cancer is currently unknown. The present study aimed to determine the possible involvement of Siva-1 in gastric cancer anticancer drug resistance in vitro. A vincristine (VCR)-resistant KATO III/VCR gastric cancer cell line with stable Siva-1 overexpression was established. The protein expression levels of Siva-1, NF-kappa B, multidrug resistance 1 (MDR1) and multidrug resistance protein 1 (MRP1) were detected via western blotting. The effect of Siva-1 overexpression on anticancer drug resistance was assessed by measuring the 50% inhibitory concentration of KATO III/VCR cells to VCR, 5-fluorouracil and doxorubicin. The rate of doxorubicin efflux and apoptosis were detected by flow cytometry. Additionally, colony formation, wound healing and Transwell assays were used to detect the proliferation, migration and invasion of cells, respectively. The results of the current study revealed that the Siva-1-overexpressed KATO III/VCR gastric cancer cells exhibited a significantly decreased sensitivity to VCR, 5-fluorouracil and doxorubicin. The results of flow cytometry revealed that the percentage of apoptotic cells decreased following overexpression of Siva-1. The colony formation assay demonstrated that cell growth and proliferation were significantly promoted by Siva-1 overexpression. Additionally, Siva-1 overexpression increased the migration and invasion of KATO III/VCR cells in vitro. Western blot analysis determined that Siva-1 overexpression increased NF-kappa B, MDR1 and MRP1 levels. The current study demonstrated that overexpression of Siva-1, which functions as a regulator of MDR1 and MRP1 gene expression in gastric cancer cells via promotion of NF-kappa B expression, inhibited the sensitivity of gastric cancer cells to certain chemotherapies. These data provided novel insight into the molecular mechanisms of gastric cancer, and may be of significance for the clinical diagnosis and therapy of patients with gastric cancer.