柚皮苷逆转人卵巢癌耐药SKOV3/DDP细胞耐药性及逆转机制的研究  

The study on drug reverse effect and reverse mechanism of naringin on multi-drug resistant human ovarian cell line SKOV3/DDP

作　　者：王磊[1] 蔡丽萍[1] 

机构地区：[1]南昌大学第一附属医院妇产科,南昌330006

出　　处：《临床肿瘤学杂志》2016年第7期598-602,共5页Chinese Clinical Oncology

基　　金：江西省科技支撑计划资助项目(15012)

摘　　要：目的 研究柚皮苷对卵巢癌顺铂（DDP）耐药细胞SKOV3/DDP体外增殖的影响及耐药逆转作用,并初步探讨相关耐药机制。方法 采用MTT法测定DDP对SKOV3和SKOV3/DDP细胞的半数抑制浓度（IC_（50））,柚皮苷对SKOV3/DDP细胞的增殖抑制作用及柚皮苷联合DDP对SKOV3/DDP细胞的增殖抑制作用。筛选出非细胞毒性的柚皮苷浓度,将实验细胞分为空白对照组、2.5μg/ml DDP组、10μmol/L柚皮苷组、10μmol/L柚皮苷联合2.5μg/ml DDP组,分别采用RTPCR和Western blotting测定各组作用48 h后耐药基因MDR1 mRNA及MRP2 mRNA的表达及其相应的表达产物P-gp、MRP2蛋白的表达水平。结果 1-32μg/ml DDP处理SKOV3及SKOV3/DDP细胞48 h后,SKOV3细胞的IC_（50）为5.48μg/ml,SKOV3/DDP细胞为14.93μg/ml,耐药指数为2.72。柚皮苷对SKOV3/DDP细胞有明显的增殖抑制作用,且呈剂量和时间依赖性。选取10μmol/L柚皮苷作为逆转耐药实验浓度,10μmol/L柚皮苷联合DDP作用48 h后,SKOV3/DDP细胞对DDP的耐药指数为1.62,逆转倍数为1.68。RT-PCR及Western blotting检测显示,柚皮苷联合DDP组与DDP单药组比较,MDR1mRNA、MRP2 mRNA表达及P-gp、MRP2蛋白表达均明显下降,差异有统计学意义（P〈0.05）。结论 柚皮苷对卵巢癌SKOV3/DDP细胞的体外增殖具有明显抑制作用,并能逆转SKOV3/DDP细胞的耐药性,其逆转耐药的机制可能与下调耐药基因MDR1 mRNA及MRP2 mRNA及相应的P-pg、MRP2蛋白的表达有关。Objective To investigate the effect of naringin on the proliferative inhibition of cisplatin （ DDP ）-resistant ovarian cancer SKOV3/DDP cells in vitro and its reverse effect of drug resistance, as well as the possible mechanisms. Methods DDP to half maximal inhibitory concentration （ IC50 ） on SKOV3 and SKOV3/DDP cells was determined by MTT method. The proliferatiive inhibi-tion of naringin alone and naringin in combination with DDP on SKOV3/DDP cells was tested by MTT method. Non-cytotoxic naringin was selected, and SKOV3/DDP cells were divided into blank control group, 2. 5μg/ml DDP group, 10μmol/L naringin group and 10μmol/L naringin＋2. 5 μg/ml DDP group. RT-PCR and Western blotting was used to detect the MDR1 and MRP2 mRNA levels and protein expressions of P-gp and MRP2. Results After treated with 1-32 μg/ml DDP for 48 h, the IC50 of SKOV3 cells and SKOV3/DDP cells were 5. 48 μg/ml and 14. 93μg/ml, and the resistance index （ RI） was 2. 72. Naringin could effectively restrain the prolif-eration of SKOV3/DDP cells in a time and dose dependent manner. The dose of 10μmol/L naringin was chosen for the further experi-ment. The RI was 1. 62 and the drug resistance reversal fold was 1. 68 times after 10μmol/L naringin＋2. 5μg/ml DDP acted on SK-OV3/DDP cells. The mRNA levels of MDR1 and MRP2, as well as the protein expressions of P-gp and MRP2 decreased significantly in 10 μmol/L naringin＋2. 5 μg/ml DDP group compared with 2. 5 μg/ml DDP group （ P〈0. 05） . Conclusion Naringin can signifi-cantly inhibit the proliferation of SKOV3/DDP cells in vitro, and can enhance the sensitivity of SKOV3/DDP cell to DDP. The reversal mechanism of drug resistance may be related to the inhibition of MDR1 and MRP2 mRNA and related proteins.

关 键 词：柚皮苷 卵巢癌 化疗耐药 SKOV3/DDP细胞 MDR1 MRP2 P-gp 

分 类 号：R737.31[医药卫生—肿瘤]