Liu W D, Sun W, Hua Y Q, Wang S G, Cai Z D
Department of Orthopedics, Shanghai General Hospital of Nanjing Medical University, Shanghai 200080, China.
Zhonghua Yi Xue Za Zhi. 2017 May 23;97(19):1510-1514. doi: 10.3760/cma.j.issn.0376-2491.2017.19.017.
To investigate the combination effect of anti-tumor agent rapamycin and anti-autophagy agent chloroquine on osteosarcoma. The inhibition effect of rapamycin or chloroquine and combination of both on human osteosarcoma cell line 143B measured with CCK-8 box.The combination index at different concentration was calculated, and the synergism effect range was analysed.The effect of combined therapy on cell cycle and apoptosis via flow cytometric was analyzted.Setting up murine subcutaneous xenograft model, the combination effect of antitumor in vivo observed. Either of them, rapamycin or chloroquine had an antitumor effect in vitro (RAPA: IC50=1.5 nmol/L; CQ: IC50=400 μmol/L). When the inhibition rate 40%<fa<70%, the combination had synergism effect (CI<1) at different concentration.Moreover, when CQ at 200 μmol/L, the synergism effect was the most obvious, CI(min)=0.255.The flow cytometric analysis display rapamycin at 1.5 nmol/L had slightly promote apoptosis effect (16.1%±4.4%), and had distinct cell cycle arrest, mainly blocked at G0/G1 phase (73.5%±10.6%; control group: 46.7%±7.7%; <0.001). While combined with chloroquine, the apoptosis cell obviously increased (21.3%±6.8%; <0.05). The xenograft model also confirmed that rapamycin combined with chloroquine had a better antitumor effect than single usage (relative tumor proliferation rate; T/C%.CQ: 98.4%±11.6%; RAPA: 65.5%±7.5%; CQ+ RAPA: 42.0%±3.4%). Rapamycin combine with chloroquine can increase the antitumor efficacy, and the combination therapy maybe a potential therapeutic regimen.
研究抗肿瘤药物雷帕霉素与抗自噬药物氯喹联合应用对骨肉瘤的作用。采用CCK-8法检测雷帕霉素、氯喹及其联合应用对人骨肉瘤细胞系143B的抑制作用。计算不同浓度下的联合指数,分析协同作用范围。通过流式细胞术分析联合治疗对细胞周期和凋亡的影响。建立小鼠皮下异种移植模型,观察体内抗肿瘤联合作用。雷帕霉素或氯喹单独应用在体外均有抗肿瘤作用(雷帕霉素:IC50 = 1.5 nmol/L;氯喹:IC50 = 400 μmol/L)。当抑制率40%<fa<70%时,联合用药在不同浓度下具有协同作用(CI<1)。此外,当氯喹浓度为200 μmol/L时,协同作用最明显,CI(min)= 0.255。流式细胞术分析显示,1.5 nmol/L的雷帕霉素有轻微促进凋亡作用(16.1%±4.4%),并使细胞周期明显阻滞,主要阻滞于G0/G1期(73.5%±10.6%;对照组:46.7%±7.7%;P<0.001)。与氯喹联合应用时,凋亡细胞明显增加(21.3%±6.8%;P<0.05)。异种移植模型也证实,雷帕霉素与氯喹联合应用的抗肿瘤效果优于单药使用(相对肿瘤增殖率;T/C%。氯喹:98.4%±11.6%;雷帕霉素:65.5%±7.5%;氯喹+雷帕霉素:42.0%±3.4%)。雷帕霉素与氯喹联合应用可提高抗肿瘤疗效,联合治疗可能是一种有潜力的治疗方案。