芸香苷逆转人大肠癌LoVo/5-氟尿嘧啶细胞耐药性及其机制  

Reversal effect of rutin on drug resistance in human colorectal cancer LoVo/5-fluorouracil cells and its mechanisms

作　　者：王婷婷 刘超怡[2] 李秀芬 李娜 刘安丽 WANG Tingting;LIU Chao-yi;LI Xiufen;LI Na;LIU An-li(Deparmen of Hisoembryoloy and Puthology,Zhengzhou Shuging Medical College,Zhengshou 450064,China;Department of Microbiology and Immunology,Huanghe Science and Technology University,Zhengzhou 450003,China;Department of Basic Medicine,He'ran Vocational Colie of Alied Tchnology,Zhengzhou 450003,China)

机构地区：[1]郑州澍青医学高等专科学校组织胚胎学与病理学教研室,郑州450064 [2]黄河科技学院微生物学免疫学教研室,郑州450003 [3]河南应用技术职业学院基础医学教研室,郑州450003

出　　处：《解剖学报》2020年第6期906-911,共6页Acta Anatomica Sinica

基　　金：河南省高等学校青年骨干教师培养项目(2017GGJS302)。

摘　　要：目的探讨芸香苷(RT)对大肠癌耐药株LoVo/5-氟尿嘧啶(5-FU)细胞耐药性的影响及其机制。方法采用浓度梯度递增法建立耐药株LoVo/5-FU细胞;用不同剂量的RT和(或) 5-FU处理48 h后,采用细胞计数试剂盒8(CCK-8)法检测细胞活力,流式细胞术检测细胞凋亡和细胞周期,RT-PCR法检测P-糖蛋白(P-gp)和多药耐药相关蛋白1(MRP1)的mRNA表达水平,Western blotting法检测P-gp、MRP1、磷酸化蛋白激酶B(p-Akt)、Akt、Survivin、细胞周期蛋白A(cyclin A)和细胞周期素依赖性蛋白激酶2 (CDK2)的蛋白表达水平。结果 LoVo/5-FU细胞中P-gp、MRP1和Survivin蛋白表达水平均显著高于LoVo细胞;5-FU和RT对LoVo/5-FU细胞的半数抑制浓度(IC50)分别为21. 77 mg/L和98. 43 mg/L;在10 mg/L RT作用下,5-FU对LoVo/5-FU细胞的IC50降至10. 64mg/L,逆转倍数为2. 05;RT可协同增强5-FU引起的LoVo/5-FU细胞凋亡和S期阻滞,抑制P-gp和MRP1基因表达以及Akt磷酸化,下调Survivin、cyclin A和CDK2蛋白水平。结论 RT可通过抑制Akt信号通路以及耐药相关蛋白的表达反转LoVo/5-FU细胞对5-FU的耐药性。Objective To investigate the effects of rutin( RT) on 5-fluorouracil( 5-FU) resistance in human colorectal cancer LoVo/5-FU cells and its possible mechanism. Methods The drug-resistant LoVo/5-FU cells were established by stepwise exposure to 5-FU. LoVo and LoVo/5-FU cells were cultured in vitro and treated with RT and( or)5-FU for 48 hours,and then the cell viability were detected by cell counting kit-8( CCK-8) assay. The cell apoptosis and cell cycle were analyzed by flow cytometry. The mRNA expression levels of P-glycoprotein( P-gp) and multidrug resistance associated protein 1( MRP1) were measured by RT-PCR. The protein levels of P-gp,MRP1,phosphorylated protein kinase B( p-Akt),Survivin,cyclin A and cyclin dependent kinase 2( CDK2) were measured by Western blotting. Results LoVo/5-FU cells had a significantly higher protein levels of P-gp,MRP1 and survivin than LoVo cells. The 50% inhibitory concentration( IC50) value of LoVo/5-FU cells to 5-FU and RT was 21. 77 mg/L and 98. 43 mg/L respectively. Under the influence of RT( 10 mg/L),the IC50 value of LoVo/5-FU cells to 5-FU reduced to 10. 64 mg/L with a reversal fold of2. 05. RT resulted in an enhance of 5-FU-induced apoptosis and S phase arrest,inactivation of Akt,and lower expression of P-gp,MRP1,survivin,cyclin A and CDK2 in LoVo/5-FU cells. Conclusion RT can reverse the drug resistance of LoVo/5-FU cells through down-regulating the expression of drug-resistant related protein and suppressing Akt signaling pathway.

关 键 词：芸香苷 5-氟尿嘧啶 大肠癌LoVo/5-FU细胞系 逆转耐药 免疫印迹法 人 

分 类 号：R735.3[医药卫生—肿瘤]