›› 2014, Vol. 35 ›› Issue (10): 1135-1135.

• 论文 • 上一篇    下一篇

卵巢癌耐药细胞株SKOV3/DDP的建立及其与凋亡途径蛋白的关系

申薇;梁冰锋;李秀荣;秦秀红;程建新   

  1. 河北医科大学第四医院妇产科,河北 石家庄,050011%河北女子职业技术学院护理系,河北 石家庄,050091
  • 发布日期:2014-10-25

SHEN Wei;LI Qinghuai;LI Xiaoyu;YAN Li;ZHANG Linlei

SHEN Wei;LIANG Bingfeng;LI Xiurong;QIN Xiuhong;CHENG Jianxin   

  • Published:2014-10-25

摘要: 目的:通过顺铂(cisplatin,DDP)诱导卵巢癌细胞珠 SKOV3建立耐药细胞株 SKOV3/DDP,并研究其与凋亡途径蛋白的关系。方法应用倒置显微镜观察 DDP 对细胞形态的影响,四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法检测 DDP 对 SKOV3和 SKOV3/DDP 细胞的增殖抑制情况,流式细胞术(flow cytometry, FCM)检测细胞凋亡率以及细胞中 X链锁凋亡抑制蛋白(X-linked inhibitor-of-apoptosis protein,XIAP)、半胱氨酸天冬氨酸蛋白酶3(cysteine-aspartic acid protease-3,Caspase-3)、B 细胞淋巴瘤/白血病2(B cell lymphoma/lewkmia-2,Bcl-2)和生存素(Survivin)的表达情况。结果 MTT法检测发现DDP作用后,SKOV3/DDP细胞的增殖抑制率较 SKOV3细胞显著降低(P<0.01),且存在着浓度和时间依赖效应(P<0.01)。由半数抑制浓度(50%concentration of inhibition,IC50)比较得 SKOV3/DDP 细胞24、48、72h 耐药指数(resistance index,RI)分别为2.434、2.950、3.780。FCM检测表明,DDP 作用后,SKOV3/DDP 凋亡率显著低于 SKOV3细胞(P<0.01)。与SKOV3细胞相比,SKOV3/DDP细胞中XIAP、Bcl-2、Survivin蛋白高表达,Caspase-3蛋白低表达,差异有统计学意义(P<0.01)。结论成功建立卵巢癌耐药细胞株 SKOV3/DDP,其耐药机制可能与 XIAP、Caspase-3、Bcl-2和Survivin蛋白的异常表达有关,表明这些蛋白参与了卵巢癌DDP耐药的形成。

关键词: 卵巢肿瘤, 顺铂, 抗药性, 肿瘤

Abstract: Objective To establish drug-resistant cell line SKOV3/DDP through ovarian cancer cell line SKOV3 by induced cisplatin (DDP ),and to study its relationship with the apoptosis pathway proteins.Methods The inverted microscope was applied to observe DDP influence on the cell morphology.Methyl thiazolyl tetrazolium (MTT)assay was used to detect proliferation inhibition rate affected by DDP on SKOV3 and SKOV3/DDP cells.Flow cytometry (FCM)was used to detect the influence of cell apoptosis and the expression of X-linked inhibitor-of-apoptosis protein (XIAP ),cysteine-aspartic acid protease-3 (Caspase-3 ),B cell lymphoma/lewkmia-2(Bcl-2)and Survivin proteins.Results MTT assay showed that proliferation inhibition rate of SKOV3/DDP cells decreased significantly compared with SKOV3 cells after DDP influence (P<0.01),and depended on the concentration and time (P<0.01).The resistance index of DDP to SKOV3/DDP and SKOV3 cells in 24h,48h and 72h were respectively 2.434,2.950 and 3.780 by comparison between 50% concentration of inhibition values.FCM revealed that the apoptosis rates of SKOV3/DDP cells to DDP decreased obviously after 48h than those of SKOV3 cells (P<0.01).Compared with SKOV3 cells,the expression of XIAP,Bcl-2 and Survivin proteins in SKOV3/DDP cells were higher,while the expression of Caspase-3 protein was lower,and their differences were statistically significant (P<0.01).Conclusion Drug-resistant cell line SKOV3/DDP for ovarian cancer is successfully established and its resistance mechanism is closely related with the abnormal expression of XIAP,Caspase-3,Bcl-2 and Survivin proteins.These proteins might be involved in the drug resistance of ovarian cancer.

Key words: ovarian neoplasms, cisplatin, drug resistance, neoplasm

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