辣椒碱抑制分化因子1表达以及核转移提高骨肉瘤对顺铂敏感性

Capsaicin enhances cisplatin sensitivity of osteosarcoma by inhibiting expression and nuclear-translocation of inhibitor of differentiation 1

目的研究辣椒碱对骨肉瘤细胞系中分化抑制因子1(Id1)的作用,并观察辣椒碱对骨肉瘤顺铂敏感性的影响。方法我院病理科收集12例骨肉瘤患者肿瘤切除组织切片以及12例骨关节炎患者全膝关节置换术后废弃的骨组织,福尔马林固定后制作石蜡切片,利用免疫组化检测Id1的表达差异。顺铂与辣椒碱共同处理MG63、HOS以及143B 3种骨肉瘤细胞系后,用免疫蛋白印迹检测Id1以及与Id1表达有关的磷酸化p65(p-p65)以及磷酸化Smad1/5(p-Smad1/5)表达改变;免疫荧光检测Id1表达部位;流式细胞学检测细胞凋亡程度。结果骨肉瘤表达高水平Id1;顺铂能介导骨肉瘤细胞株Id1表达上调(P<0.05)以及Id1入核;辣椒碱通过抑制p65以及Smad1/5磷酸化抑制Id1蛋白表达(P<0.05);联合应用辣椒碱能抑制顺铂引起Id1入核(P<0.05)并加强凋亡效果(P<0.05)。结论辣椒碱能通过抑制顺铂介导的骨肉瘤Id1上调以及核转移,加强顺铂介导骨肉瘤细胞凋亡效果,可作为潜在的化疗辅助药物。

Objective To investigate the expression of inhibitor of differentiation 1（ Id1） in osteosarcoma tissues and evaluate the effect of capsaicin to enhance the chemosensitivity of osteosarcoma to cisplatin. Methods Immunohistochemistry was used to investigate Id1 expression levels in 12 osteosacroma tissue specimens and bone tissues from 12 patients with osteoarthritis. Three osteosarcoma cell lines（ MG63,HOS and 143 B） were treated with cisplatin and capsaicin,alone or in combination,and the expression levels of Id1,p-p65 and p-Smad1/5 in the cells were detected with Western blotting; immunofluorescence assay was used to investigate the location of Id1,and flow cytometry was used to analyze cell apoptosis in the treated cells. Results Osteosarcoma tissues expressed high levels of Id1 compared with osteoarthritis tissues. In the osteosarcoma cell lines,cisplain treatment induced an obvious up-regulation of Id1 expression（ P〈0. 05） and nuclear translocation of Id1,while treatment with capsaicin significantly reduced Id1 expression by suppressing p65 and Smad1/5（ P〈0. 05）. In cells treated with both capsaicin and cisplatin,capsaicin obviously reduced cisplatin-induced nuclear translocation of Id1（ P〈0. 05） and significantly enhanced cisplatin-mediated cell apoptosis（ P〈0. 05）. Conclusion Capsaicin can inhibit cisplatin-induced up-regulation and nucleartranslocation of Id1 and enhance cisplatin-mediated osteosarcoma cell apoptosis,and may thus serve as a potential agent for adjuvant chemotherapy of osteosarcoma.