Suppression of heat shock protein 70 by siRNA enhances the antitumor effects of cisplatin in cultured human osteosarcoma cells
Suppression of heat shock protein 70 by siRNA enhances the antitumor effects of cisplatin in cultured human osteosarcoma cells
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DOI：
10.1007/s12192-017-0793-x
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发表时间：
2017-09-01
影响因子：
3.8
通讯作者：
Kubo, Toshikazu
中科院分区：
文献类型：
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作者：
Mori, Yuki;Terauchi, Ryu;Kubo, Toshikazu
尽管化疗的进步改善了骨肉瘤的预后，但一些患者对治疗的反应并不充分。热休克蛋白70 （Hsp70）在癌细胞中高水平表达，降低抗癌药物的治疗效果，导致预后较差。本研究探讨了小干扰RNA （siRNA）介导的骨肉瘤细胞系Hsp70表达抑制是否会增强对顺铂的敏感性。采用Western blotting和实时逆转录聚合酶链反应（RT-PCR）观察顺铂治疗后Hsp70的表达情况。观察siRNA抑制Hsp70后顺铂IC50的变化。通过检测末端脱氧核苷酸转移酶介导的dUTP镍端标记（TUNEL）、caspase-3活性和线粒体膜电位来评估顺铂诱导细胞凋亡的有效性。Hsp70的表达上调依赖于顺铂的浓度。抑制Hsp70表达可显著降低顺铂的IC50。在抑制Hsp70表达的骨肉瘤细胞中加入顺铂后，TUNEL、caspase-3和线粒体膜电位检测显示细胞凋亡显著增加。抑制Hsp70表达可诱导培养骨肉瘤细胞凋亡，表明抑制Hsp70可增强对顺铂的敏感性。抑制Hsp70的表达可能为骨肉瘤提供一种新的辅助治疗方法。
Although advances in chemotherapy have improved the prognosis for osteosarcoma, some patients do not respond sufficiently to treatment. Heat shock protein 70 (Hsp70) is expressed at high levels in cancer cells and attenuates the therapeutic efficacy of anticancer agents, resulting in a poorer prognosis. This study investigated whether small interfering RNA (siRNA)-mediated inhibition of Hsp70 expression in an osteosarcoma cell line would enhance sensitivity to cisplatin. The expression of Hsp70 with cisplatin treatment was observed by using Western blotting and real-time reverse transcription polymerase chain reaction (RT-PCR). Changes in the IC50 of cisplatin when Hsp70 was inhibited by siRNA were evaluated. Cisplatin's effectiveness in inducing apoptosis was assessed by assay of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), caspase-3 activity, and mitochondrial membrane potential. Up-regulation of Hsp70 expression was dependent on the concentration of cisplatin. Inhibition of Hsp70 expression significantly reduced the IC50 of cisplatin. When cisplatin was added to osteosarcoma cells with Hsp70 expression inhibited, a significant increase in apoptosis was demonstrated in TUNEL, caspase-3, and mitochondrial membrane potential assays. Inhibition of Hsp70 expression induced apoptosis in cultured osteosarcoma cells, indicating that Hsp70 inhibition enhanced sensitivity to cisplatin. Inhibition of Hsp70 expression may provide a new adjuvant therapy for osteosarcoma.