低频超声联合微泡对比剂通过PI3K/AKT通路对乳腺癌耐药株MCF-7/ADR多药耐药的逆转机制研究  

Low-frequency ultrasound combined with microbubble contrast agent reversed the multidrug resistance of breast cancer resistant strain MCF-7/ADR through PI3K/AKT pathway

作　　者：贺修宝 胡小梅[2] 郭雅婧[1] HE Xiu-bao;HU Xiao-mei;GUO Ya-jing(Department of Ultrasound,Chenzhou First People's Hospital,Chenzhou 423000,China;Medical Record Room,Chenzhou First People's Hospital,Chenzhou 423000,China)

机构地区：[1]郴州市第一人民医院超声科,423000 [2]郴州市第一人民医院病案室,423000

出　　处：《天津医药》2021年第8期802-807,共6页Tianjin Medical Journal

基　　金：湖南省卫健委课题(202109020028)。

摘　　要：目的探索低频超声联合微泡(LFUSMB)通过磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路对乳腺癌耐药株MCF-7/阿霉素(ADR)多药耐药的逆转效果及机制。方法CCK-8法筛选LFUSMB作用时间并测定ADR对MCF-7/ADR细胞的半抑制浓度(IC_(50))。将MCF-7/ADR细胞分组为:对照(C)组,采用MCF细胞培养基正常培养;ADR组,采用加入ADR(终质量浓度20 mg/L)的MCF细胞培养基培养;LFUSMB+ADR组,采用加入ADR(终质量浓度20 mg/L)的MCF细胞培养基培养并经LFUSMB处理30 s;LFUSMB+ADR+740 YP(PI3K/AKT通路活化剂)组,采用加入ADR(终质量浓度20 mg/L)和740 YP(终质量浓度50 mg/L)的MCF细胞培养基培养并经LFUSMB处理30 s。Annexin V-FIFC/PI法检测MCF-7/ADR细胞凋亡率,Western blot检测P糖蛋白(P-gp)、抗多药耐药蛋白(MRP)1、MRP2、乳腺癌抗性蛋白(BCRP/ABCG2)、PI3K、AKT、p-PI3K、p-AKT蛋白表达。结果采用LFUSMB干预30 s进行后续研究。LFUSMB 30 s组ADR对MCF/ADR细胞的IC_(50)低于未超声处理组,相对耐药逆转倍数约为8.20倍。ADR组细胞凋亡率较C组增高(P<0.05),MRP1、MRP2、P-gp、BCRP/ABCG2、p-PI3K/PI3K和p-AKT/AKT蛋白水平无明显变化;LFUSMB+ADR组细胞凋亡率较ADR组增高,MRP1、MRP2、P-gp、BCRP/ABCG2、p-PI3K/PI3K和p-AKT/AKT蛋白水平显著下调(P<0.05)。与LFUSMB+ADR组比较,LFUSMB+ADR+740 YP组细胞凋亡率显著降低,MRP1、MRP2、P-gp、BCRP/ABCG2蛋白水平显著增高(P<0.05)。结论LFUSMB可通过抑制PI3K/AKT通路活化,下调MRP1等腺苷三磷酸结合盒(ABC)家族蛋白表达,逆转乳腺癌MCF-7/ADR细胞耐药性。Objective To explore the reversal effect and mechanism of low-frequency ultrasound combined with microbubbles(LFUSMB)on the multidrug resistance of breast cancer resistant strains MCF-7/Adriamycin(ADR)through the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway.Methods The CCK-8 method was used to screen the action time of LFUSMB and determine the half inhibitory concentration(IC_(50))of ADR on MCF-7/ADR cells.The MCF-7/ADR cells were divided into:control group(normally cultured with MCF cell culture medium),ADR group(used MCF cell culture medium with ADR,final concentration 20 mg/L),LFUSMB+ADR group(cultured with MCF cell culture medium with ADR,final concentration 20 mg/L and treated with LFUSMB for 30 s)and LFUSMB+ADR+740 YP(PI3K/AKT pathway activator)group(MCF cell culture medium added with ADR,final concentration 20 mg/L and 740 YP,final concentration 50 mg/L,and were cultured and treated with LFUSMB for 30 s).Annexin V-FIFC/PI method was used to detect the apoptosis rate of MCF-7/ADR cells.Western blot assay was used to detect the expression of glycoprotein(P-gp),multidrug resistance protein(MRP)2,breast cancer resistance protein(BCRP/ABCG2),PI3K,AKT,p-PI3K,p-AKT and MRP1 proteins.Results The LFUSMB intervention for 30 s was used for the follow-up study.The IC_(50) of ADR to MCF/ADR cells was lower in the LFUSMB 30 s group than that of the non-ultrasound group,and the relative drug resistance reversal ratio was approximately 8.20 times.The apoptosis rate of MCF-7/ADR cells was significantly higher in the ADR group than that in the control group(P<0.05).There were no significantly differences in the protein levels of MRP1,MRP2,P-gp,BCRP,p-PI3K/PI3K and p-AKT/AKT between the two groups.The apoptosis rate of MCF-7/ADR cells was higher in the LFUSMB+ADR group than that in the ADR group(P<0.05),and the protein levels of MRP1,MRP2,P-gp,BCRP,p PI3K/PI3K and p-AKT/AKT were significantly lower than those in the ADR group(P<0.05).Compared with the LFUSMB+ADR group,the cell apoptosis rate was significa

关 键 词：乳腺肿瘤 细胞凋亡 超声疗法 MCF-7细胞 P糖蛋白 阿霉素 低频超声联合微泡 PI3K/AKT通路 

分 类 号：R737.9[医药卫生—肿瘤]