慢病毒介导的HER2-siRNA对人卵巢癌移植瘤的抑制及其显像监测  

Growth inhibition of human ovarian cancer by lentivirus-mediated HER2-siRNA monitored with radioimmunoimaging

作　　者：钱红[1] 吴强乐[2] 王小侠[3] 杨仪[1] 尤嘉熙[1] 朱本兴[4] 石怡珍[1] 

机构地区：[1]215004,苏州大学附属第二医院核医学科 [2]杭州师范大学附属医院放射科 [3]215004,苏州大学附属第二医院肿瘤科 [4]江苏省放射医学与防护重点实验室、苏州大学医学部放射医学与防护学院

出　　处：《中华核医学与分子影像杂志》2013年第6期-,共5页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：江苏省放射医学与防护重点实验室开放课题，苏州市科技发展计划(社会发展-应用基础研究)

摘　　要：目的 研究慢病毒介导的干扰RNA沉默人表皮生长因子受体2(HER2)对卵巢癌SKOV-3细胞移植瘤生长的影响,并探讨放射免疫显像在RNA干扰肿瘤生物治疗中的应用价值.方法 利用HER2-短发夹状(sh) RNA慢病毒表达载体和随机序列慢病毒载体感染SKOV-3细胞株,建立稳定表达HER2-小干扰(si) RNA的SKOV-3细胞株及阴性对照(NC)细胞株.分别将细胞株接种于裸鼠,建立荷人卵巢癌裸鼠模型实验(KD1)组和NC组,并以未感染的SKOV-3细胞株裸鼠模型作为空白对照(CON)组,检测各组裸鼠(每组5只)的成瘤率、成瘤时间、瘤体大小、HER2蛋白表达.通过131Ⅰ-曲妥珠单克隆抗体(Herceptin)进行荷瘤小鼠放射免疫显像,观察肿瘤显影情况,比较各实验组T/B比值.采用SPSS 17.0对实验结果进行单因素方差分析和最小显著差异t检验.结果 裸鼠成瘤率为100％(15/15).CON组、NC组和KD1组的平均成瘤时间分别为(4.583±0.520)、(4.567±0.284)和(6.023±0.316) d(F=13.946,P＜0.01),KD1组移植瘤较另2组平均成瘤时间延长(t=4.557、4.608,均P＜0.01).至种植后28 d各组间移植瘤体积差异有统计学意义(F=26.343,P＜0.01);CON组、NC组和KD1组离体瘤质量分别为(0.614±0.135)、(0.558±0.190)和(0.120±0.489) g(F=225.026,P＜0.01);KD1组平均瘤体积(t=7.125、4.759)、瘤体质量(t=19.158、16.977)较另2组明显减小(均P＜0.01).免疫组织化学结果显示KD1组HER2蛋白表达明显减少(弱阳性).核素显像示各组荷瘤小鼠移植瘤均有不同程度显影,KD1组1、4、8、12、24、48、72和96 h T/B比值(0.208 ～4.427)均低于NC组(0.576～5.508)和CON组(0.640～5.695;F=9.197～ 39.375,均P＜0.05).结论 慢病毒介导的HER2-siRNA可以显著抑制人卵巢癌移植瘤的生长;131Ⅰ-Herceptin放射免疫显像能在一定程度上反映HER2蛋白的表达情况,有望用于针对HER2靶点的RNA干扰肿瘤生物治疗的疗效评估.Objective To investigate the effect of human epidermal growth factor receptor-2 (HER2) silenced by lentivirus-mediated RNA interference (RNAi) on the growth of SKOV-3 ovarian cancer,and to explore the value of radioimmunoimaging in monitoring the biotherapy of RNAi.Methods The ovarian cancer cell line SKOV-3 was infected with lentivirus-mediated HER2-short hairpin (sh) RNA expression vector and scrambled control lentivirus vector,respectively.Both infected cells were inoculated into nude mice to establish two ovarian cancer xenograft models:knock down 1 (KD1) group and normal control (NC) group.The uninfected SKOV-3 xenograft model served as blank control (CON) group.The tumor formation rate,tumor generation time and tumor size at different time points were measured.The expression of HER2 protein was measured by immunohistochemistry.1n Ⅰ-Herceptin was injected before radioimmunoimaging,and the T/B ratios were acquired.One-way analysis of variance and the least significant difference (LSD)-t test were performed with SPSS 17.0.Results All mice models were constructed successfully (100％,15/15).The average time of tumor generation was (4.583±0.520) d,(4.567±0.284) d and (6.023±0.316) d in CON,NC and KD1 groups,respectively(F=13.946,P＜0.01).The tumor formation time of KD1 group was significantly longer than the other two groups (t=4.557,4.608,both P＜0.01),respectively.On the 28th day after the tumor cell implantation,the tumor size was significantly different among the three groups (F=26.343,P＜0.01).The tumor mass was (0.614±0.135) g,(0.558±0.190) g and (0.120±0.489) g in CON,NC and KD1 groups,respectively (F=225.026,P＜0.01).Both the tumor size (t=7.125,4.759) and tumor mass (t=19.158,16.977) of KD1 group were significantly less than those of CON and NC groups (all P＜0.01),respectively.Immunohistochemical results showed that the HER2 protein expression was inhibited in the KD1 group.The tumor could be visualized clearly on radioimmunoimaging at different time points.The T/B ratio of the KD1 group (

关 键 词：卵巢肿瘤 肿瘤移植 受体 表皮生长因子-尿抑胃素 RNA干扰 

分 类 号：R817.5[医药卫生—影像医学与核医学] R737.33[医药卫生—放射医学]