葡萄糖调节蛋白78在卵巢癌细胞衰老过程中的作用  

Role of GRP78 Protein in the Senescence of Ovarian Cancer Cells

作　　者：李伟[1] 李双[1] 李莉[1] 贾瑶[1] 杨洁[1] 栗妍[1] 汪辉[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《华中科技大学学报（医学版）》2014年第4期391-394,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81001152)

摘　　要：目的拟了解葡萄糖调节蛋白78(GRP78)在卵巢癌细胞衰老过程中所发挥的作用,并初步进行相关机制探讨。方法亚凋亡剂量顺铂诱导卵巢癌A2780细胞衰老,并继续培养至出现衰老逃脱现象;衰老相关的!-半乳糖苷酶检测细胞衰老情况;流式细胞仪分析细胞周期情况;Western blot检测GRP78及相关蛋白表达情况。siRNA反义抑制GRP78的表达,观察衰老逃脱细胞克隆形成情况。结果亚凋亡剂量顺铂作用A2780细胞后发生明显G2/M期阻滞,细胞衰老染色阳性。继续培养3周后出现衰老逃脱细胞克隆的形成,并可被顺铂再次诱导进入衰老状态。GRP78蛋白在衰老的A2780细胞及再次被顺铂诱导进入衰老状态的衰老逃脱细胞中均呈低表达,而在衰老逃脱细胞中重新恢复其高表达。反义抑制A2780细胞中GRP78蛋白表达后P53及Cyclin B1蛋白表达均明显升高,Cdc2蛋白表达明显下降,同时伴有细胞衰老逃脱能力的明显降低。结论 GRP78在顺铂诱导卵巢癌细胞衰老过程中起到抵抗衰老的作用。Objective Chemotherapeutic drugs of sub-apoptosis dose can induce the senescence-like change of tumor cells and the senescence-escaped phenomenon.This study was designed to investigate the role of GRP78 protein in the senescence of ovarian cancer cells and the related mechanism.Methods Cisplatin of sub-apoptosis dose was used to induce the senescence of A2780 cells,which were then cultured to become senescence-escaped cells.Senescence-associatedβ-galactosidase was used to detect cell senescence.Cell cycle was detected by flow cytometry.The expression levels of GRP78 and its related proteins were measured by Western blot.The formation of senescence-escaped cell clone was observed after the GRP78 expression was knocked down by siRNA.Results After treatment with cisplatin of sub-apoptosis dose,A2780 cells were obviously arrested at G2/M phase and positive for senescence-associatedβ-galactosidase.Senescence-escaped cell clones were formed after culture for3 weeks and they could be induced into senescence again by cisplatin.The GRP78 protein was lowly expressed in senescent A2780 cells and senescence-escaped cells that were re-induced into senescence by cispaltin,but retrieved to a high level of expression in senescence-escaped cells.The expression levels of P53 and Cyclin B1 protein were significantly increased,and the expression level of Cdc2 protein and the senescence-escaped capability decreased obviously in A2780 cell with GRP78 protein knocked down by siRNA.Conclusion GRP78 protein plays a anti-senescence role during cisplatin-induced senescence in ovarian cancer cells.GRP78 may be used as a new target to improve the efficacy of chemotherapy for ovarian cancer.

关 键 词：葡萄糖调节蛋白78 卵巢癌 衰老逃脱细胞 P53 

分 类 号：R737.31[医药卫生—肿瘤]