尼妥珠单抗联合顺铂对人舌鳞状细胞癌细胞株CAL-27的抑制作用  

Inhibitory effects of nimotuzumab combined with DDP on human tongue squamous carcinoma cell line CAL-27

作　　者：王秀力[1] 孟箭[2] 

机构地区：[1]徐州市中心医院中心实验室,221009 [2]徐州市中心医院口腔科,221009

出　　处：《中华口腔医学研究杂志（电子版）》2013年第1期13-17,共5页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：徐州市科技发展项目(XZZD1017)

摘　　要：目的研究尼妥珠单抗(h-R3)联合顺铂(DDP)对人舌鳞状细胞癌(以下简称舌鳞癌)细胞株CAL-27的生长抑制作用。方法常规培养CAL-27细胞,以5×104/ml的细胞密度种植细胞,实验分为对照组、h-R3组、DDP组及h-R3联合DDP用药组共4组。CCK-8法检测h-R3及h-R3联合DDP组在不同时间对CAL-27细胞的生长抑制情况;分别于24、48及72h对各组进行摄片并收集细胞培养上清,酶联免疫吸附法(ELISA)检测h-R3组及h-R3联合DDP组在不同时间对CAL-27细胞分泌表皮生长因子(EGF)及血管内皮细胞生长因子(VEGF)含量的影响,同时采用细胞凋亡-Hoechst染色试剂盒检测CAL-27加入不同药物后的凋亡情况。结果 h-R3和DDP单药对CAL-27细胞生长均有抑制作用且呈时间和剂量依赖性,两药单独作用72h后,对CAL-27细胞的最大抑制率分别为(26.91±7.08)%和(89.18±4.73)%。两药联用对CAL-27细胞增殖的最大抑制率为(93.26±1.03)%,联合用药可提高细胞增殖抑制率,呈相加作用;h-R3及h-R3联合DDP组不同时间凋亡率与对照组比较差异有统计学意义(P<0.01),随着作用时间增加细胞凋亡率亦呈升高趋势;h-R3及h-R3联合DDP组不同时间CAL-27细胞分泌EGF与VEGF含量与对照组比较均显著降低(P<0.01),而且联合用药与单药比较,EGF和VEGF含量也显著降低(P<0.05)。结论 h-R3在体外与DDP联用可以提高对CAL-27细胞增殖抑制及凋亡作用,同时对CAL-27细胞分泌EGF与VEGF具有抑制作用。Objective To study the inhibitory effects of nimotuzumab combined with DDP on human tongue squamous carcinoma Cell Line CAL-27. Methods CAL-27 cells were cultured by the density of 5 x 104/ml. H-R3 group, DDP group, h-R3 ＋DDP group and control group containing 10% FBS were used to study the inhibitory effects of h-R3 combined with DDP on CAL-27 ceils. CCK-8 assay was performed to evaluated the growth inhibitory ratio of CAL-27 cells in the different time. After CAL-27 cells were cultured for 24, 48 and 72： h, the supernatant liquid of which were collected and used for enzyme-linked immunosorbent assay （ELISA） to detect the content of EGF and VEGF. Different drug alone or combination effect on CAL-27 cells apoptosis changes were determined by Hoechst Staining Kit. Results H-R3 and DDP both inhibited the growth of CAL-27 ceils in a time and dose- dependent manner. The single drug of h-R3 or DDP had effect on CAL-27 cells, after 72 hour treatment, the proliferation inhibition ratio of CAL-27 cells was （26.91 ± 7.08）% and （89.18 ± 4.73）% seperately. Combination of h-R3 and DDP could increase the proliferation inhibition ratio of CAL-27 cells, which was （93.26 ± 1.03）% after 72 hour treatment, and the two drugs had synergistic role on the proliferation inhibition. After the drug h-R3 and DDP alone or combined treatment, the apoptosis ratioof combination group was increased, and the apoptosis ratio was increased when treatment time prolonged. The content of EGF and VEGF in h-R3 and DDP group were obviously degraded than that in control group （P 〈 O.01）, and the content of EGF and VEGF in h-R3＋DDP group were obviously degraded than that in b-R3 or DDP group （P 〈 0.05）. Conclusiort H-R3, DDP and h-R3 combined with DDP couM significantly inhibit cell line CAL-27 proliferation and the secretion of EGF and VEGF, also could induce cell apoptosis, and the effect was in accordance with concentration and time-dependent.

关 键 词：尼妥珠单抗 顺铂 舌鳞状细胞癌细胞株 

分 类 号：R739.8[医药卫生—肿瘤]