临床肿瘤学杂志

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蟾毒灵对人骨肉瘤细胞化疗增敏作用的实验研究

张剑军1,沙婧婧1,胡海燕1,何爱娜1,王 兵2,孙元珏1,沈 赞1,姚 阳1   

  1. 1 200233 上海 上海交通大学附属第六人民医院肿瘤内科 2 200233 上海交通大学附属第六人民医院中医内科
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2013-10-31 发布日期:2013-10-31
  • 通讯作者: 姚 阳

Experiment of bufalin on chemosensitivity to human osteosarcoma cells in vitro

ZHANG Jianjun, SHA Jingjing, HU Haiyan, HE Aina, WANG Bing, SUN Yuanjue, SHEN Zan, YAO Yang   

  1. Department of Internal Oncology, Sixth Peoples Hospital, Shanghai JiaoTong University, Shanghai 200233, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-10-31 Published:2013-10-31
  • Contact: YAO Yang

摘要:

目的 探讨蟾毒灵对人骨肉瘤细胞Saos2和U2OS的化疗增敏作用。方法 采用无毒剂量的蟾毒灵联合不同浓度的阿霉素(0.01、0.1、1.0μg/ml ADM)和顺铂(0.5、1.0、2.0μg/ml DDP)分别作用于Saos-2和U2OS细胞24h,用CCK-8法观察药物对细胞增殖的抑制作用,Hoechst 33258 染色法观察细胞凋亡的形态学改变,流式细胞术检测细胞凋亡率。
结果CCK-8法检测显示,蟾毒灵对Saos2和U2OS细胞增殖的抑制作用呈浓度依赖性;无细胞毒剂量(0.005μmol/L)的蟾毒灵可以显著增强不同浓度ADM和DDP 对Saos-2和U2OS
细胞增殖的抑制作用(q>1.15)。Hoechst 33258染色法显示,蟾毒灵联合DDP或ADM后Saos-2和U2OS细胞的凋亡形态学变化较DDP或ADM单药更显著。流式细胞术检测显示,0.005μmol/L蟾毒灵联合1.0μg/ml DDP或2.0μg/ml ADM组Saos-2和U2OS细胞的凋亡率显著高于DDP或ADM单药组(q>1.15,P<0.05)。结论 无毒剂量的蟾毒灵可以增强化疗药物诱导的人骨肉瘤细胞凋亡的能力,具有化疗增敏作用。

Abstract:

Objective To investigate the effect of bufalin on chemosensitivity to human osteosarcoma cells Saos-2 and U2OS in vitro. Methods Saos-2 and U2OS cells were treated with bufalin at nontoxic dosage in combination with 0.01, 0.1, 1.0μg/ml doxorubicin(ADM) or 0.5, 1.0, 2.0μg/ml cisplatin(DDP), cell proliferation was determined by CCK-8 assay, the morphological changes of cell apoptosis were observed by Hoechst 33258 staining, and apoptosis was assessed by flow cytometric analysis. Results CCK-8 assay showed that proliferation inhibition of bufalin on human osteosarcoma cells Saos-2 and U2OS was in a dose-dependent manner, and 0.005μmol/L bufalin in combination with ADM or DDP could enhance proliferation inhibition of Saos-2 and U2OS cells compared with ADM or DDP alone(q>1.15). Hoechst 33258 staining indicated that bufalin combined with ADM or DDP showed more obvious morphological changes of apoptosis in cells than ADM or DDP alone. Flow cytometric analysis showed that the apoptosis rates of 0.005μmol/L bufalin combined with 1.0μg/ml DDP or 2.0μg/ml ADM was significantly higher than those of cells treated with ADM or DDP alone(q>1.15, P<0.05).
ConclusionBufalin at non-toxic dosage can enhance ADM or DDP-induced cell apoptosis of human osteosarcoma cells, showing effect of chemotherapy
sensitization.

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