拓扑特肯诱导人口腔上皮癌细胞凋亡的机制_医学论文.docVIP

拓扑特肯诱导人口腔上皮癌细胞凋亡的机制_医学论文 拓扑特肯诱导人口腔上皮癌细胞凋亡的机制_医学论文 作者：徐强，王枫，李等松，陈长生，徐文，缪珊 【关键词】 人口腔上皮癌细胞株 　　【Abstract】 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 TPTtreated KB cells and Westernblot was used to test the phosphorylations of p38MAPK proteins. RESULTS: The inhibition ratio of TPTtreated 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 (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 (0.01). Agarose gel electrophoresis of genomic DNA of TPTtreated 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, 0.05). CONCLUSION: TPTinduced apoptosis of KB cells in vitro may result from the enhanced activation of p38MAPK. 　　【Keywords】 TPT(Topotecan) KB cells phosphorylated protein38 apoptosis 　　【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30 mg/L的TPT孵育12, 24和48 h,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，Westernblot检测pp38MAPK的表达. 结果: TPT处理细胞12, 24和48 h后,KB 细胞的抑制率分别为(24±9)%, (34±7)%和(45±10)%,各组间有显著性差异(0.05)；流式细胞仪检测其凋亡率，12 h时(11.5±2.8)%， 24 h( 31.2±4.1)%，各组间有显著性差异(0.01)；DNA凝胶电泳可见TPT作用12 h组DNA碎片化，是典型的细胞凋亡生化指标；TPT处理细胞12, 24和48 h后,KB细胞的pp38MAPK表达逐渐升高(n=3, 0.05). 结论: TPT可通过pp38MAPK开放表达信号通路诱导体外培养的TPT 处理KB细胞凋亡. 　　【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 　　0引言 　　拓扑特肯(Topotecan, TPT)为9二甲基氨基10羟基喜树，是近年来临床上用于治疗晚期肺癌的新型化疗药物,在美国已被批准用于晚期难治性卵巢癌［1］. 目前国内对TPT的研究报道较少,其抗肿瘤作用机制尚未明确. 丝裂原活化的蛋白激酶（MAPKs）在将细胞外刺激信号转导至细胞及核内的过程中，通过丝/苏/酪氨酸磷酸化，参与细胞增殖、分化、转化及凋亡等及其重要的生物学反应，其中p38 MAPK