灵芝多糖逆转卵巢癌细胞株顺铂耐药的作用及其机制  

Effect of reversion of ganoderma lucidum polysaccharides on cisplatin resistant in ovarian cancer cells and its mechanism

作　　者：曲红光[1,2,3] 高磊[1,2] 贺丹[1,2] 刘聪[4] 燕巍[1,2] 高嵩[1,2] 王丽[1,2] 

机构地区：[1]吉林大学白求恩医学院病原生物学教研室 [2]吉林大学真菌研究中心,吉林长春130021 [3]吉林省长春市妇产医院生殖中心,吉林长春130042 [4]大连医科大学基础医学院,辽宁大连116044

出　　处：《吉林大学学报（医学版）》2011年第2期250-254,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划重点项目资助课题(20080444-2)

摘　　要：目的:探讨灵芝多糖(GLPS)对卵巢癌耐药细胞SKOV-3/DDP耐药性的逆转作用,阐明GLPS通过改善卵巢癌耐药基因而改善其耐药的机制。方法:用不同剂量梯度(4、40和400mg·L-1)GLPS、顺铂(DDP,1、10和100mg·L-1)作用卵巢癌耐药细胞SKOV-3/DDP,用MTT法每12h检测细胞抑制率(共检测72h),得到最佳抑制浓度后,将实验用细胞分为空白组、GLPS组和DDP组,应用实时PCR技术检测相关耐药基因GSTM1、GSTT1和GSTP1的表达,应用Western blotting技术检测相关耐药蛋白GSTM1的表达。结果:40和400mg·L-1的GLPS作用SKOV-3/DDP细胞12～72h,SKOV-3/DDP细胞生长抑制,且其抑制率随着时间的延长和浓度的升高而增加(P<0.05),最佳抑制浓度为40mg·L-1。浓度为10和100mg·L-1 DDP作用SKOV-3/DDP细胞后细胞生长无明显变化(P>0.05)。40mg·L-1 GLPS和10mg·L-1 DDP作用SKOV-3/DDP细胞36h,GSTM1、GSTT1和GSTP1的基因表达水平均下降(P<0.05),GSTM1蛋白表达水平明显下降。结论:GLPS能抑制耐药基因GSTM1、GSTT1和GSTP1的表达,改善肿瘤细胞的耐药性,增强肿瘤细胞对化疗药物的敏感性。Objective To investigate the reversal effect of ganoderma lucidum polysaccharides（GLPS） on drug resistance in ovarian cancer cells SKOV-3/DDP and clarifyits drug-resistance mechanism of GLPS through improving the resistance genes of ovarian cancer.Methods The ovarian cancer cells SKOV-3/DDP were treated with different concentration gradients of GLPS（4,40 and 400 mg·L-1） and cisplatin（1,10 and 100 mg·L-1）,the inhibitory rates were detected by MTT method per 12 h（total test 12-72 h）,the experimental cells were divided into control group,GLPS group and cisplatin group when the best inhibitory concentration was gotten.The expressions of related resistantce genes GSTM1,GSTT1 and GSTP1 were detected by real-time PCR.The expression differences of GSTM1-related resistance protein were detected by Western blotting method.Results The inhibitory rates of SKOV-3/DDP cells were higher after treated with 40 and 400 mg·L-1 GLPS for 12-72 h,and the inhibitory rate was increased with the time prologation and the increase of concentration（P〈0.05）,and the best inhibitory concentration was 40 mg·L-1.10 and 100 mg·L-1 cisplatin didn’t have effect on the inhibitory rate of SKOV-3/DDP cells（P〉0.05）.The expressons of GSTM1,GSTT1 and GSTP1 of SKOV-3/DDP cells were decreased after treated with 40 mg·L-1 GLPS and 10 mg·L-1 cisplatin for 36 h（P〈0.05）.The expression of GSTM1 protein was decreased the most.Conclusion GLPS can inhibit the expressions of drug resistance genes GSTM1,GSTT1 and GSTP1,and improve the resistance effect of tumor cells and enhance their sensitivityies to chemotherapeutic drugs.

关 键 词：SKOV-3/DDP 灵芝多糖 耐药基因 

分 类 号：R285.5[医药卫生—中药学] R737.31[医药卫生—中医学]