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72 h 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72 h 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 × 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 cells. 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 cells in a time and dosedependent 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 ratio of 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＜0.01），and the content of EGF and VEGF in h-R3+DDP group were obviously degraded than that in h-R3 or DDP group （P＜0.05）.

Conclusion

H-R3，DDP and h-R3 combined with DDP could 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.