CAJ | 학술논문

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b-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,其可作为一个提高卵巢癌化疗疗效的新靶点进行进一步的研究。
목적：의료해포도당조절단백78(GRP78)재란소암세포쇠로과정중소발휘적작용,병초보진행상관궤제탐토。방법아조망제량순박유도란소암 A2780세포쇠로,병계속배양지출현쇠로도탈현상；쇠로상관적b-반유당감매검측세포쇠로정황；류식세포의분석세포주기정황；Western blot 검측 GRP78급상관단백표체정황。siRNA 반의억제GRP78적표체,관찰쇠로도탈세포극륭형성정황。결과아조망제량순박작용 A2780세포후발생명현 G2/M기조체,세포쇠로염색양성。계속배양3주후출현쇠로도탈세포극륭적형성,병가피순박재차유도진입쇠로상태。GRP78단백재쇠로적 A2780세포급재차피순박유도진입쇠로상태적쇠로도탈세포중균정저표체,이재쇠로도탈세포중중신회복기고표체。반의억제 A2780세포중 GRP78단백표체후P53급Cyclin B1단백표체균명현승고,Cdc2단백표체명현하강,동시반유세포쇠로도탈능력적명현강저。결론 GRP78재순박유도란소암세포쇠로과정중기도저항쇠로적작용,기가작위일개제고란소암화료료효적신파점진행진일보적연구。
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 de-tect 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 for 3 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 ex-pression in senescence-escaped cells.The expression levels of P5 3 and Cyclin B1 protein were significantly increased,and the ex-pression 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.