反义寡核苷酸并超声微泡造影剂转染联合超声照射逆转肝癌多药耐药

Reversion of multidrug resistance of hepatocellular carcinoma by antisense oligonucleotides and ultrasonic microbubble intensifier transfection combined with ultrasound irradiation

目的探讨体内、体外mdr1、mrp、lrp反义寡核苷酸(AODNs)并超声微泡造影剂转染联合低强度超声照射逆转肝癌多药耐药的可行性,寻找逆转肿瘤多药耐药有效和靶向的方法。方法利用超声微泡造影剂包载肿瘤耐药基因mdr1、mrp、lrp的AODNs进行转染,联合低强度超声照射,以肝癌细胞多药耐药细胞模型(HepG2／ADM) 为研究对象,通过逆转录聚合酶链反应、western blot和四甲基偶氮唑盐法,从体外细胞培养及动物实验,研究 AODNs并超声微泡造影剂转染联合超声照射逆转癌细胞多药耐药及降低肿瘤恶性表型和成瘤能力的作用。结果 HepG2／AMD细胞增殖被抑制,其mdr1和mrp的mRNA、蛋白质表达水平明显降低;裸鼠皮下移植瘤生长受抑制。结论体外、体内AODNs并超声微泡造影剂转染联合低强度超声照射能有效逆转人肝癌细胞HepG2／ADM的多药耐药, 该技术可能为肝癌临床治疗提供新的思路。

Objective To study whether antisense oligonucleotides and ultrasonic microbubble intensifier transfection combined with ultrasound irradiation is an effective and directional way in reversing multidrug resistance （MDR） in tumors, Methods Mdr1, mrp, and lrp genes antisense oligonucleotides on the ultrasound microbubble intensifier were transfected for the human HepG2/ADM cell lines and then the cells were radiated with low intensity ultrasound, The effects of the reversion of carcinoma cells＇ MDR and the reduction of their malignancy and growth capability in vitro and in vivo were assessed using RTPCR, Western blot and MTT. Results The treatment restrained the multiplication of the human HepG2/AMD cell lines. The levels of their mRNA and protein of cells＇ mdr1 and mrp genes dropped significantly. Growth of the subcutaneous transplanted tumors in the nude mice decreased. Conclusions Transfection of MDR genes antisense oligonucleotides on the ultrasonic microbubble intensifier combined with low intensity ultrasound radiation may serve as a new treatment method for hepatocellular carcinoma.