mTOR特异性抑制剂RAD001对SGC7901/DDP细胞耐药的逆转作用  

Inhibitor of mTOR: RAD001 can reverse drug resistance of SGC7901 /DDP

作　　者：刘莹[1] 朱祖安[2] 孔庆兖[1] 刘磊[1] 崔涛[1] 张瑞瑞[1] 

机构地区：[1]徐州医学院病理学教研室,徐州221000 [2]徐州医学院附属医院消化内科,徐州221000

出　　处：《临床与实验病理学杂志》2013年第11期1206-1210,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：江苏省卫生厅课题(Z201016);江苏省六大人才高峰第七批次项目资助(032)

摘　　要：目的观察哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)特异性抑制剂RAD001对胃癌SGC7901/DDP细胞顺铂耐药逆转的作用,并初步探讨其发挥作用的机制。方法常规培养胃癌SGC7901/DDP细胞,单独RAD001、DDP(cisplatin,顺铂)或联合药物作用后,流式细胞术检测细胞周期,免疫细胞化学和Western blot技术检测药物作用后细胞中P-gp、MRP1、p27蛋白表达。结果单独DDP组(顺铂组)作用于SGC7901/DDP细胞后,细胞周期无明显改变,与对照组相比,差异无统计学意义(P>0.05)。RAD001组(RAD分别为2.5、5、10 nmol/L)、联合用药组(RAD 2.5 nmol/L+DDP 2.5 mg/L)作用48 h后,处于G1期细胞比例增加,与对照组相比,差异有统计学意义(P<0.05);处于S期细胞比例降低,与对照组相比,差异有统计学意义(P<0.05);联合用药组效果强于单独用药组(P<0.05);免疫组化检测结果示:顺铂组对细胞中上述蛋白的表达无明显改变作用。RAD预处理后,RAD001组、联合用药组作用48 h后细胞中P-gp和MRP1表达下降,p27表达升高,与对照组相比,差异有统计学意义(P<0.05)。联合用药组作用效果强于单独用药组(P<0.05);Western blot检测示:顺铂组对细胞中上述蛋白的表达无明显改变。RAD预处理后,RAD001组、联合用药组作用48 h后细胞中P-gp和MRP1表达均下降,与对照组相比,差异均具有统计学意义(P<0.05)。p27表达均升高,与对照组相比,差异有统计学意义(P<0.05)。联合用药组作用强于各单独药物作用组(P<0.05)。结论 RAD001能够逆转胃癌细胞SGC7901/DDP耐药,其可能通过上调p27表达引起细胞周期阻滞而发挥作用。Purpose To investigate the role and the mechanism of RAD001 on reverse drug-resistant of SGC7901/DDP. Methods SGC7901/DDP cells were cultivated and treated with DDP, RAD001 alone or RAD001 in combination with DDP. Flow cytometry, im- munocytochemistry and Western blot assay were used to investigate cell cycle and protein expressions of P-gp, MRP1, p27 in cells. Result The group of DDP 2.5 mg/L alone showed no effect of cell cycle in SGC7901/DDP cells compared with control group （ P 〉 0. 05 ）. The group of RAD001 （2. 5, 5, 10 nmol/L） and the group of combination （ RAD 2. 5 nmol/L ＋ DDP 2.5 mg/L） showed that the percentage of cells in G1 phase increased （ P 〈 0. 05 ）, however, the percentage of cells in S phase decreased （ P 〈 0. 05 ）. The group of combination is better than the groups of single drug. The results of immunohistochemistry showed that there＇ s no change of a- bove proteins in the group of DDP. After pretreatment with RAD for 48 h, the expression of P-gp （ P 〈 0. 05 ） and MRP1 （ P 〈 0. 05 ） in RAD001 group, combination group decreased and the expression of p27 increased （P 〈 0. 05 ）. The combined group was better than single drug group （ P 〈 0. 05 ）. The result of Western blot test showed that there＇ s no change of above proteins in the group of DDP. After pretreatment with RAD for 48 h, P-gp and MRP1 expression were decreased in RAD001 group, combination group （ P 〈 0. 05 ）. The expression of p27 were increased （P 〈 0. 05 ）. The combined group was stronger than the single drug group （ P 〈 0.05 ）. Conclu- sion RADO01 could reverse drug-resistant of SGC7901/DDP through up-regulating p27 and inducing the arrest of cell cycle in GI phase.

关 键 词：RAD001 细胞周期 SGC7901 DDP细胞 耐药逆转 

分 类 号：R735.2[医药卫生—肿瘤]