洛铂对鼻咽癌抗肿瘤作用的体外实验研究

The anticancer effect of lobaplatin on human nasopharyngeal carcinoma cells in vitro

目的探讨洛铂对鼻咽癌的体外抗肿瘤作用及其可能的机制,为洛铂用于鼻咽癌的临床治疗提供实验依据。方法在体外将不同浓度洛铂分别作用于低分化鼻咽癌细胞株CNE2、HONE1和SUNE1,采用CCK-8法检测细胞活性;碘化丙啶(PI)染色分析细胞中DNA的含量,流式细胞仪检测细胞周期情况;采用Annexin V-FITC/PI双染法检测细胞凋亡情况;采用蛋白印迹法(Westernblotting)检测细胞周期和细胞凋亡相关蛋白的表达。结果在体外洛铂作用3种鼻咽癌细胞株48h具有明显的细胞抑制作用,并表现出浓度依赖性。洛铂作用鼻咽癌细胞株CNE2、HONE1、SUNE1的半数抑制浓度(IC50)分别为(4.05±0.49)μmol/L、(4.32±1.17)μmol/L和(2.51±0.15)μmol/L。洛铂作用3种鼻咽癌细胞株48h后,在较低浓度(0.125倍IC50)时将细胞周期阻滞在G2期,同时伴随G2期相关蛋白CyclinB1和phospho-cdc2(Tyr15)表达增高;而在较高浓度(0.5倍IC50)时则诱导细胞呈Caspase依赖性细胞凋亡,并具有浓度依赖性。结论洛铂对鼻咽癌细胞具有明显的细胞毒作用,且呈浓度依赖性;其机制表现为双重作用,即在较低浓度时阻滞细胞于G2期和在较高浓度时诱导细胞凋亡,这为洛铂用于鼻咽癌的临床治疗提供可能性。

Objective To investigate the anticancer effect of lobaplatin on human nasopharyngeal carcinoma（NPC） in vitro and the probable mechanisms to determine whether lobaplatin is a candidate for chemotherapy in human NPC. Methods Three kinds of poorly differentiated human NPC cell lines CNE2, HONE1 and SUNE1 were engaged in this experiment. Cytotoxicity of lobaplatin in the cell lines was examined with CCK-8 cell viability assay. Cell cycle and apoptosis were determined by flow cytometry and the expres- sions of its related proteins were examined by Western blotting analysis. Results Lobaplatin with different concentration showed evi- dent inhibitory effect on the three kinds of poorly differentiated human NPC cell lines and there was increased in a concentration de- pendent manner. The proliferation of human NPC cell lines CNE2, HONE1 and SUNE1 were inhibited when they were treated with lobaplatin for 48h. The half maximal inhibitory concentration（ ICs0 ） of three kinds of NPC cell lines CNE2, HONE1 and SUNE1 were （4. 05 ±0. 49） μmol/L, （4. 32± 1.17） μmol/L and （2. 51 ±0. 15） μmol/L,respectively. Lobaplatin arrested cell cycle progression in G2 phase with increasing cyclin B1 and phospho-cdc2 （Tyr15） proteins in lower doses, however, induced apoptosis with caspase-3 cleaved in higher doses. Conclusion Lobaplatin shows anticaneer effect on human NPC cells by means of cell cycle blockage and induction of apoptosis, which will contribute to the potential use of lobaplatin in the treatment of human NPC.