蛋白激酶B调控的p53蛋白直接线粒体转移与卵巢癌细胞顺铂耐药的关系  

Relationship between the Akt.regulated direct p53 mitochondrial translocation and the resistance tocisplatin of ovarian cancer cells

作　　者：周莹[1] 王树玉[1] 杨晓葵[1] 王红梅[2] 

机构地区：[1]首都医科大学附属北京妇产医院生殖医学科,100026 [2]中国科学院动物所生殖生物学国家重点实验室

出　　处：《中华肿瘤杂志》2011年第2期97-100,共4页Chinese Journal of Oncology

基　　金：国家自然科学基金（30872746）;北京市自然科学基金（7082034）;北京市优秀人才基金（20061D0301000091）

摘　　要：目的探讨蛋白激酶B（PKB，又称Akt）调控的p53蛋白直接线粒体转移在顺铂诱导的卵巢癌细胞凋亡中的作用及与卵巢癌顺铂耐药的关系。方法应用Western blot检测顺铂作用前后卵巢癌顺铂敏感细胞0V2008、A2780s和顺铂耐药细胞C13^＊、A2780cp中线粒体和全细胞中p53含量。分离纯化线粒体，检测卵巢癌敏感细胞与耐药细胞p53直接线粒体作用的差异；应用Hoechst染色法检测卵巢癌细胞的凋亡率，将Aktl／2siRNA导入耐药细胞C13^＊细胞中，观察p53线粒体转移作用和细胞凋亡率的改变。结果顺铂能引起p53进入0V2008和A2780s细胞的线粒体中，但不能进入C13^＊和A2780cp细胞的线粒体。过度表达活化的Akt2能抑制p53进入A2780s细胞的线粒体；下调Akt后，可使p53进人C13^＊细胞的线粒体，同时C13^＊细胞对顺铂的敏感性增加。结论顺铂能引起卵巢癌敏感细胞中p53转移至线粒体，Akt导致的卵巢癌化疗耐药在一定程度上是Akt对p53直接线粒体作用的抑制。下调Akt的表达能使卵巢癌化疗耐药细胞的p53直接线粒体作用增强，耐药性得到一定程度的逆转。Objective To explore Akt-regulated direct p53 mitochondrial translocation in cisplatin- induced apoptosis in ovarian cancer cells and the relationship between this and chemoresistance in ovarian cancer. Methods Chemosensitive ovarian cancer cell lines （0V2008 and A2780s） and chemoresistant cells （C13 ＊ and A2780cp） were treated with cisplatin and whole cell and mitochondrial p53 contents were determined by Western blot. The p53 accumulation in mitochondria was determined in purified mitochondrial fractions in cisplatin-sensitive and -resistant ovarian cancer cells. Aktl/2 siRNA were transfected into C13 ＊ cells. Cisplatin-induced apoptosis was measured by Hoechst staining and p53 translocation was determined by Western blot. Results Cisplatin induced mitochondrial p53 accumulation and apoptosis in chemosensitive cells （ P 〈 0.05 ）, but not in resistant cells （ P 〉 0.05 ）. Over-expression of active Akt2 inhibited p53 directly translocate to mitoehondria, and downregulation of Akt by Aktl/2 siRNA increased p53 mitochondrial accumulation and sensitize C13 ＊ cells to cisplatin treatment. Conclusions Cisplatin induces direct p53 mitochondrial accumulation in chemosensitive cells, and Akt confers resistance in ovarian cancer cells, in part, by regulating the direct action of p53 in mitochondrial death pathway.

关 键 词：蛋白激酶B P53蛋白 卵巢肿瘤 细胞 0V2008 细胞 A2780s 抗药性 肿瘤 

分 类 号：R737.31[医药卫生—肿瘤]