拓扑特肯诱导人口腔上皮癌细胞凋亡的机制  

Mechanism of apoptosis of KB cells induced by Topotecan (TPT)

作　　者：徐强[1] 王枫[2] 李等松[3] 陈长生[4] 徐文[5] 缪珊[6] 

机构地区：[1]兰州铁路局兰西铁路医院口腔科,甘肃兰州730050 [2]第四军医大学预防医学系营养与食品卫生学教研室,陕西西安710033 [3]兰州军区军事医学研究所 [4]第四军医大学预防医学系统计学教研室 [5]第四军医大学预防医学系毒理学教研室 [6]第四军医大学科研部药研所

出　　处：《第四军医大学学报》2004年第16期1474-1477,共4页Journal of the Fourth Military Medical University

摘　　要：目的 :研究拓扑特肯 (Topotecan ,TPT)诱导体外培养的人口腔上皮癌细胞 (KB)凋亡的作用及其可能机制 .方法 :体外培养KB细胞 ,加 30mg/L的TPT孵育 1 2 ,2 4和4 8h ,用MTT法检测TPT对KB细胞增殖的作用 ;DNA凝胶电泳和流式细胞仪技术检测TPT诱导KB细胞凋亡 ,West ern blot检测 p p38MAPK的表达 .结果 :TPT处理细胞 1 2 ,2 4和 4 8h后 ,KB细胞的抑制率分别为 (2 4± 9) % ,(34±7) %和 (4 5± 1 0 ) % ,各组间有显著性差异 (P <0 .0 5 ) ;流式细胞仪检测其凋亡率 ,1 2h时 (1 1 .5± 2 .8) % ,2 4h(31 .2± 4 .1 ) % ,各组间有显著性差异 (P <0 .0 1 ) ;DNA凝胶电泳可见TPT作用 1 2h组DNA碎片化 ,是典型的细胞凋亡生化指标 ;TPT处理细胞 1 2 ,2 4和 4 8h后 ,KB细胞的 p p38MAPK表达逐渐升高 (n =3,P <0 .0 5 ) .结论 :TPT可通过p p38MAPK开放表达信号通路诱导体外培养的TPT处理KB细胞凋亡 .AIM: To study the mechanism of apoptosis of KB cells induced by Topotecan (TPT). METHODS: MTT assay was applied to test the proliferation arrest of KB cells under the treatment of TPT for 12, 24 and 48 h, agarose gel electrophoresis of genomic DNA and flow cytometry (FCM) were used to examine the apoptosis of TPT treated KB cells and Western blot was used to test the phosphorylations of p38MAPK proteins. RESULTS: The inhibition ratio of TPT treated KB cells for 12, 24 and 48 h was (24.4± 9.1)%, (33.7±6.6)% and (45.1±10.4)%, respectively, with significant difference between different treatment duration ( P <0.05). By FCM, the apoptosis rate of the cells was (11.5± 2.8)% at 12h and (31.2±4.1)% at 24 h, with significant difference ( P <0.01). Agarose gel electrophoresis of genomic DNA of TPT treated KB cells for 12 h showed DNA fragmentations, typical index of apoptosis in biochemistry. The phosphorylations of p38 increased gradually with the prolongation of TPT treatment (12, 24 and 48 h) ( n=3, P <0.05). CONCLUSION: TPT induced apoptosis of KB cells in vitro may result from the enhanced activation of p38MAPK.

关 键 词：拓扑特肯 人口腔上皮癌细胞株 磷酸化的丝裂原活化蛋白激酶38 凋亡 

分 类 号：R739.8[医药卫生—肿瘤]