Department of Gastrointestinal and Peripheral Vascular Surgery, People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China.
Department of Surgery, People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China.
Cell Biochem Biophys. 2020 Dec;78(4):455-463. doi: 10.1007/s12013-020-00929-y. Epub 2020 Jul 9.
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.
SIVA-1 在许多不同的细胞系中诱导细胞凋亡中发挥关键作用，并参与顺铂（DDP）介导的抗肿瘤作用的机制。然而，SIVA-1 参与胃癌顺铂耐药性尚未被揭示。为了探讨 SIVA-1 对 DDP 耐药性的影响，构建了重组 pGV358-GFP-SIVA-1 慢病毒载体，并转染人顺铂耐药 MKN45/DDP 胃癌细胞。随后，在 MKN45/DDP 细胞中建立了稳定的 SIVA-1 过表达，导致 MKN45/DDP 细胞对 DDP 的敏感性增加。流式细胞术表明，与对照组相比，SIVA-1 过表达增加了凋亡细胞的百分比。集落形成实验清楚地表明，SIVA-1 过表达后细胞生长和增殖明显受到抑制。此外，SIVA-1 过表达抑制了 MKN45/DDP 细胞在体外的迁移和侵袭能力。Western blot 分析表明，SIVA-1 增加了 p53、p73 和 p14ARF 的表达水平，而降低了 Bcl-2、MDM2 和 Bcl-xL 的表达。总之，SIVA-1 上调了 p53、p73 和 p14ARF 的蛋白表达，下调了 Bcl-2、MDM2 和 Bcl-xL 的表达，在体外逆转了胃癌细胞的顺铂耐药性，表明 SIVA-1 可能成为减轻化疗耐药性的有价值的潜在靶点。
