中性微泡、阳离子微泡及靶向阳离子微泡携内皮抑素基因经超声转染治疗裸鼠乳腺癌种植瘤的实验研究

Study of endostatin gene therapy on breast cancer in nude mice by ultrasoundtargeted microbubbles destruction with netural microbubbles,cationic microbubbles and targeted cationic microbubbles

目的研究中性微泡(NMB)、阳离子微泡(CMB)及靶向阳离子微泡(CMB105)携内皮抑素(ES)质粒在超声作用下转染治疗MDA-MB-231细胞裸鼠种植瘤,观察其对肿瘤模型生长的影响及治疗作用。方法 18只裸鼠均注射MDA-MB-231细胞悬液建立种植瘤模型,随机分为3组(NMB组、CMB组及CMB105组),每组6只。NMB、CMB及CMB105携ES质粒在超声作用下转染MDA-MB-231种植瘤,每只裸鼠左侧肿瘤接受治疗,右侧肿瘤为对照。治疗前和治疗后每3 d观察记录肿瘤生长情况,绘制肿瘤生长曲线;治疗后15 d处死裸鼠,剥取肿瘤计算抑瘤率,行荧光定量聚合酶链反应和蛋白质免疫印迹法分别检测Endostatin m RNA及蛋白表达情况。结果于转染后12 d开始,CMB组和CMB105组治疗侧肿瘤生长均受到抑制。转染后15 d,CMB105组治疗侧抑瘤率为(53.18±5.69)%,明显高于CMB组治疗侧[(27.57±3.02)%]和NMB组治疗侧[(10.63±1.47)%],差异均有统计学意义(均P<0.05)。CMB组和CMB105组ES m RNA及蛋白表达增加,两组比较差异有统计学意义(P<0.01)。结论携载ES质粒的CMB105在超声作用下其抗肿瘤治疗效应明显优于携载ES质粒的CMB及NMB,可在一定程度上延缓裸鼠种植瘤生长。

Objective To study the therapeutic effects on breast cancer in nude mice by ultrasound -targeted microbubbles destruction（UTMD） with netural microbubbles（NMB）,cationic microbubbles（CMB） and CMB105 carrying endostatin（ES） plasmid.Methods MDA-MB-231 tumor cell suspensions were injected subcutaneously in 18 nude mice,they were randomly divided into three groups（NMB group,CMB group and CMB 105 group） with six mice in each group. Tumor xenografts on the left hind limb received the ultrasound mediated gene therapy,and the right hind limb tumor xenografts were used as control.The size of tumor and the curve of tumor growing were recorded before and after treatment every 3 d.15 d after treatment, the mice were sacrificed,the tumor growth inhibition rate was calculated and the tumor growth curve was delineated. qPCR and Western-blot were used to evaluate the level of ES mRNA and protein.Results UTMD-mediated ES gene delivery both with CMB and CMB105 significantly inhibited tumor growth from 12 d. 15 d after transfer,the inhibition rates of CMB105,CMB and NMB group were （53.18±5.69）%,（27.57±3.02）% and （10.63±1.47）%,respectively,there was significant difference （all P〈0.05）. The level of ES mRNA and protein were increased both in CMB and CMB105 group,there was significant difference （P 〈0.01）.Conclusion UTMD -mediated ES gene delivery with CMB105 can inhibit the tumor growth successfully,and the therapy effects is better than CMB and NMB group.