摘要
背景与目的:肿瘤坏死因子相关凋亡诱导配体(tumornecrosisfactor-relatedapoptosisinducingligand,TRAIL)可选择性杀伤肿瘤细胞,而不影响正常细胞生长。当一部分肿瘤细胞对TRAIL不敏感时,特定的其它药物可增强其杀伤作用。本文旨在探讨结肠癌细胞SW480对TRAIL的敏感性,以及TRAIL与阿霉素联用对细胞的杀伤作用及可能作用机制。方法:常规培养结肠癌细胞SW480。利用MTT法检测细胞毒性作用,流式细胞术定量分析凋亡细胞比例,透射电镜在亚细胞结构形态上证实凋亡细胞,Westernblot分析p53及bcl-2蛋白表达变化。结果:(1)SW480细胞对TRAIL不敏感,100ng/mlTRAIL只能杀伤7.8%的细胞,IC50>1000ng/ml,且不存在浓度依赖性。(2)SW480细胞对阿霉素敏感,存在浓度依赖性作用,IC50=65μmol/L,0.86μmol/L的阿霉素对细胞不表现杀伤作用。(3)TRAIL与阿霉素合用表现出协同作用,亚毒性浓度TRAIL(100ng/ml)与亚毒性浓度阿霉素(0.86μmol/L)联用可杀伤80%SW480细胞。流式细胞学证实这种杀伤作用主要通过诱导细胞凋亡实现,透射电镜亦观察到大量凋亡细胞存在。药物作用前后,p53及bcl-2蛋白表达水平无明显改变。结论:结肠癌细胞株SW480对TRAIL不敏感,但TRAIL与亚毒性浓度阿霉素联用对癌细胞有协同杀伤作用,这种细胞毒性作用主要表现?
BACKGROUND &OBJECTIVE:Tumor necrosis factor related apoptosis inducing ligand (TRAIL) could selectively kill tumor cells. This effect could be improved using some therapeutic agents. The aim of this study was to determine the sensitivity of SW480 cells to TRAIL, the interaction of TRAIL and doxorubicin against colon cancer cells and its possible mechanism. METHODS: SW480 cells were cultured with RPMI1640 medium in regular condition. Cytotoxicity was examined by MTT assay. Cell apoptosis was detected by flow cytometry. The subcellular morphology was observed by electron microscopy. The changes of p53 and Bcl 2 in protein level were quantified by Western blot analysis. RESULTS:(1)SW480 cells were not sensitive to TRAIL,and the IC50 was more than 1 000 ng/ml. 100 ng/ml of TRAIL could only kill 7 8%of the cells. (2)Concentration dependent cytotoxicity of doxorubicin was exhibited in SW480 cells, with IC50 of 65 μmol/L. 0 86 μmol/L of doxorubicin did not affect cell growth. (3)The combination of TRAIL and doxorubicin exhibited synergistic effect on SW480 cells. Subtoxic TRAIL(100 ng/ml)and subtoxic doxorubicin(0 86 μmol/L) killed 80%of the cells. The synergistic cytotoxicity was large partly attributed to cell apoptosis, which was proved by simultaneous flow cytometry assay and electron microscopy. However, TRAIL and doxorubicin did not affect p53 and Bcl 2 protein expression. CONCLUSION: TRAIL in combination with subtoxic doxorubicin could effectively kill SW480 cells, which did not respond to TRAIL alone. Apoptosis was the main manner of this killing effect, and the apoptotic pathway induced by TRAIL and doxorubicin did not involved in the change of p53 and Bcl 2 protein expression.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第8期816-820,共5页
Chinese Journal of Cancer
关键词
TRAIL
阿霉素
结肠癌
细胞SW480
协同杀伤作用
细胞凋亡
Tumor necrosis factor related apoptosis inducing ligand (TRAIL)
Doxorubicin
Synergistic effect
Apoptosis
