吴茱萸碱诱导人宫颈癌HeLa细胞凋亡过程中非caspase调控因素

Intracellular regulation besides caspase of evodiamine-induced cell death in HeLa cells

目的 研究吴茱萸碱 (evodiamine)诱导人宫颈癌HeLa细胞凋亡机制中非caspase调控因素。方法 MTT法 ,WesternBlot免疫印迹法 ,流式细胞术。结果 虽吴茱萸碱对HeLa细胞杀伤作用介于化疗药物放线菌素D (actino mycineD)、顺铂 (cisplatin)和 5 氟尿嘧啶 (5 fluorouracil,5 Fu)之间 ,但对细胞撤除药物后的继续增殖能力的影响相似。MAP激酶 (MAPK)抑制剂中ERK和p38的抑制剂不影响细胞死亡 ,而JNK抑制剂 (SP6 0 0 12 5 )可协同吴茱萸碱杀死细胞 ,但吴茱萸碱并未影响JNK及磷酸化JNK蛋白表达。吴茱萸碱可将HeLa细胞周期阻滞在G2 /M期 ,发生大量凋亡。结论 对JNK激酶的抑制可协同促进吴茱萸碱对HeLa细胞的杀伤作用 ,而且吴茱萸碱可将细胞周期阻滞在G2 /M期发生凋亡。

AIM To study non caspase pathway in evodiamine induced cell death in HeLa cells. METHODS MTT assay, Western Blot analysis, and flow cytometric analysis were used. RESULTS Compared to actinomycin D, cisplatin and 5 Fu, evodiamine had less cytotoxic effects on HeLa cells, but evodiamine induced similar inhibition of cell growth in HeLa cells which were incubated with evodiamine for 24 h, then cultured with free drug medium for two weeks. ERK and p38 MAPKs was not involved in evodiamine induced cell death. However, JNK inhibitor (SP600125) made cells more sensitive to evodiamine. JNK and phosphorylated JNK expression were not influenced by evodiamine. Moreover, evodiamine induced cell cycle arrest at G 2/M phase. CONCLUSION Evodiamine induces the G 2/M arrest. The inhibition of JNK exaggerates evodiamine induced cell death in HeLa cells.