超声辐照维拉帕米微泡体外对神经母细胞瘤多药耐药的逆转作用  

Ultrasound irradiation microbubble verapamil reverses multidrug resistance of neuroblastoma cells in vitro

作　　者：陈镜宇[1] 全学模[1] 郭振华[2] 王荞[1] 唐毅[1] 王佚[2] 

机构地区：[1]重庆医科大学附属儿童医院超声科,重庆400014 [2]重庆医科大学附属儿童医院普通外科,重庆400014

出　　处：《第三军医大学学报》2010年第16期1724-1727,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30600579);重庆市教委科学技术研究项目(市教委KJ060304)~~

摘　　要：目的探讨超声辐照维拉帕米(verpamil,VER)微泡逆转肿瘤多药耐药的作用及机制。方法制备包裹维拉帕米的乳酸-羟基乙酸共聚物(VER-PLGA)超声微泡造影剂;通过长春新碱(VCR)多次小剂量递增诱导法建立神经母细胞瘤耐药株(SK-N-SH/VCR),并检测其耐药性;根据处理因素不同分为6组:A:空白对照组(SK-N-SH/VCR培养组),B:SK-N-SH/VCR+VER(50μg/ml)组,C:SK-N-SH/VCR+VER(250μg/ml)组,D:SK-N-SH/VCR+超声辐照组,E:SK-N-SH/VCR+PLGA微泡+超声辐照组,F:SK-N-SH/VCR+VER-PLGA微泡(VER50μg/ml)+超声辐照组。48h后,利用RT-PCR和免疫组织化学染色法(ICC)检测各组细胞中MDR1基因及P-gp的表达水平,评估VER在不同处理组中逆转肿瘤细胞耐药的效果。结果①成功制备包裹维拉帕米的VER-PLGA微泡造影剂,包封率为32%;②成功建立神经母细胞瘤耐药株SK-N-SH/VCR,MTT法证实其对不同化疗药物的耐药倍数是野生株的2.3～5.4倍;③ICC证实耐药株高表达MDR1基因;④RT-PCR及免疫组化法检测发现MDR1基因在F组中的光密度值及P-gp表达水平较其他组明显降低(P<0.05)。结论超声辐照包裹小剂量维拉帕米的微泡可有效逆转肿瘤多药耐药。Objective To study the effects of ultrasound irradiation microbubbles with verpamil （VER） on reversing tumor multidrug resistance.Methods VER-PLGA microbubbles were prepared with double emulsion and freeze drying techniques.The neuroblastoma cell line SK-N-SH/VCR with multidrug resistance （MDR） phenotype was established by inducing with small-doses of VCR.Then the cells were divided into 6 groups according to different treatments,that is,group A：SK-N-SH/VCR,group B：SK-N-SH/VCR＋VER （50 μg/ml）,group C：SK-N-SH/VCR＋VER （250 μg/ml）,group D：SK-N-SH/VCR＋ultrasound,group E：SK-N-SH/VCR＋PLGA＋ultrasound,group F：SK-N-SH/VCR＋VER-PLGA （50 μg/ml）＋ultrasound.RT-PCR and immunocytochemical staining were used to detect the expression levels of MDR1 gene and P-gp respectively.And the effects of VER on reversing multidrug resistance of tumor in different groups were evaluated.Results The VER-PLGA microbubbles were prepared successfully,and the encapsulated rate was 32%.SK-N-SH/VCR cells with MDR phenotype were established and the resistance index （RI） was 2.3 to 5.4 times to different chemotherapy drugs when compared with the wild cells.Compared with others,the level of MDR1 gene was the most lowest in group F （P0.05）.The expression levels of P-gp and MDR1 gene were the lowest in group F among the all 6 groups （P0.05）.Conclusion Ultrasound irradiation microbubbles enhance the effects of verapamil to reverse the multidrug resistance of tumor.

关 键 词：超声 微泡 维拉帕米 肿瘤 多药耐药 

分 类 号：R739.4[医药卫生—肿瘤] R972.2[医药卫生—临床医学]