Lentivirus-Mediated Overexpression of SIVA-1 Reverses Cisplatin Resistance in Gastric Cancer in vitro
Lentivirus-Mediated Overexpression of SIVA-1 Reverses Cisplatin Resistance in Gastric Cancer in vitro
复制标题
慢病毒介导的 SIVA-1 过表达可逆转胃癌的体外顺铂耐药性
DOI：
10.1007/s12013-020-00929-y
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发表时间：
2020-07-09
影响因子：
2.6
通讯作者：
Mai, Wei
中科院分区：
文献类型：
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作者：
Wang, Xiao-Tong;Li, Lei;Mai, Wei
SIVA-1在许多不同细胞系的凋亡诱导中起关键作用，并参与顺铂（DDP）介导的抗肿瘤作用的机制。然而，SIVA-1在胃癌顺铂耐药中的参与尚未被揭示。为探讨SIVA-1对顺铂耐药的影响，构建重组慢病毒载体pGV 358-GFP-SIVA-1，转染人顺铂耐药胃癌细胞MKN 45/DDP。随后，在MKN 45/DDP细胞中建立了稳定的SIVA-1过表达，这导致MKN 45/DDP细胞体外顺铂敏感性增加。流式细胞术显示，与对照组相比，SIVA-1过表达增加了凋亡细胞的百分比。殖民地测定清楚地显示，在SIVA-1过表达后，细胞生长和增殖被显著抑制。此外，SIVA-1过表达抑制MKN 45/DDP细胞的体外迁移和侵袭能力。Western blot分析表明SIVA-1增加了p53、p73和p14 ARF的表达水平，而减少了Bcl-2、MDM 2和Bcl-xL的表达。总之，SIVA-1在体外上调p53、p73和p14 ARF的蛋白表达，并降低Bcl-2、MDM 2和Bcl-xL的蛋白表达，随后逆转胃癌细胞中的顺铂耐药性，这表明SIVA-1作为减轻化疗耐药性的有价值的潜在靶点。
SIVA-1 plays a critical role in the induction of apoptosis in a number of different cell lines and participates in the mechanism of cisplatin (DDP)-mediated antitumor effects. However, the involvement of SIVA-1 in cisplatin resistance in gastric carcinoma has not been revealed. To explore the effect of SIVA-1 on DDP resistance, a recombinant pGV358-GFP-SIVA-1 lentiviral vector was constructed and transfected into human cisplatin-resistant MKN45/DDP gastric cancer cells. Subsequently, stable SIVA-1 overexpression was established in MKN45/DDP cells, which resulted in increased DDP sensitivity in MKN45/DDP cells in vitro. Flow cytometry demonstrated that SIVA-1 overexpression increased the percentage of apoptotic cells compared to that in the control. The colony formation assay clearly revealed that cell growth and proliferation were significantly suppressed following SIVA-1 overexpression. In addition, overexpression of SIVA-1 inhibited the migratory and invasive potential of MKN45/DDP cells in vitro. Western blot analysis indicated that SIVA-1 increased the expression levels of p53, p73, and p14ARF, whereas it reduced Bcl-2, MDM2, and Bcl-xL expression. In short, SIVA-1 upregulated the protein expression of p53, p73, and p14ARF and decreased that of Bcl-2, MDM2, and Bcl-xL in vitro and subsequently reversed cisplatin resistance in gastric cancer cells, suggesting that SIVA-1 serves as a valuable potential target for attenuating chemotherapy resistance.