Luo Leiming, Wang Yuanhui, Yin Yiran, Ge Jianhua, Lu Xiaobo
Department of Bone and Joint Surgery, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, P.R. China.
Oncol Lett. 2018 Mar;15(3):3548-3551. doi: 10.3892/ol.2018.7730. Epub 2018 Jan 4.
The effects of NIN1/RPN12 binding protein 1 homolog (NOB1) on the pathogenesis of osteosarcoma and its expression on the chemosensitivity to cisplatin were investigated. Seventy-four patients with osteosarcoma who received surgical resection in The Affiliated Hospital of Southwest Medical University (Sichuan, China) from September 2013 to September 2016 were enrolled in this study. The expression of NOB1 in cancer and cancer-adjacent tissues of patients was detected by reverse transcription-polymerase chain reaction, and the relationship between NOB1 expression and the pathogenesis of osteosarcoma was analyzed. The expression of NOB1 in osteosarcoma MG-63 cells was interfered with using small interfering ribonucleic acid (siRNA). Western blotting was used to detect the transfection efficiency and changes in apoptosis indicators. Cell Counting Kit-8 (CCK-8) assay was used to examine changes in the sensitivity of cells to cisplatin. The effect of NOB1 knockout on cell apoptosis was examined by flow cytometry. In patients with osteosarcoma, the level of NOB1 mRNA in cancer tissues was significantly higher than that in cancer-adjacent tissues (p<0.05), and the expression of NOB1 was correlated with Ennecking staging and tumor size (p<0.05). The expression level of the apoptotic indicator caspase-3 was activated after siRNA interfered with NOB1 expression, thus reducing the expression level of anti-apoptotic indicator B-cell lymphoma 2. CCK-8 results showed that the downregulation of NOB1 increased the sensitivity of MG-63 cells to cisplatin (p<0.05). In addition, flow cytometry showed that the downregulation of NOB1 significantly promoted the apoptosis of MG-63 cells. NOB1 is significantly upregulated in patients with osteosarcoma, thus reducing the curative effect of cisplatin chemotherapy, which indicates that the prognosis is poor.
研究了NIN1/RPN12结合蛋白1同源物(NOB1)对骨肉瘤发病机制的影响及其对顺铂化疗敏感性的表达。本研究纳入了2013年9月至2016年9月在西南医科大学附属医院(中国四川)接受手术切除的74例骨肉瘤患者。通过逆转录-聚合酶链反应检测患者癌组织和癌旁组织中NOB1的表达,并分析NOB1表达与骨肉瘤发病机制的关系。使用小干扰核糖核酸(siRNA)干扰骨肉瘤MG-63细胞中NOB1的表达。采用蛋白质免疫印迹法检测转染效率和凋亡指标的变化。使用细胞计数试剂盒-8(CCK-8)检测细胞对顺铂敏感性的变化。通过流式细胞术检测NOB1基因敲除对细胞凋亡的影响。在骨肉瘤患者中,癌组织中NOB1 mRNA水平显著高于癌旁组织(p<0.05),且NOB1表达与Enneking分期和肿瘤大小相关(p<0.05)。siRNA干扰NOB1表达后,凋亡指标caspase-3的表达水平被激活,从而降低了抗凋亡指标B细胞淋巴瘤-2的表达水平。CCK-8结果显示,NOB1的下调增加了MG-63细胞对顺铂的敏感性(p<0.05)。此外,流式细胞术显示,NOB-1的下调显著促进了MG-63细胞的凋亡。骨肉瘤患者中NOB1显著上调,从而降低了顺铂化疗的疗效,提示预后不良。