磁感应加热和HSV—tk自杀基因及核素内照射联合治疗乳腺癌的实验研究  

Combined anti-tumor therapeutic effect of targeted gene, hyperthermia, radionuclide brachytherapy in breast carcinoma

作　　者：陈道桢[1] 唐秋莎[2] 项静英[1] 许飞[1] 张立[1] 王峻峰[1] 

机构地区：[1]南京医科大学附属无锡妇幼保健院中心实验室,214002 [2]东南大学基础医学院病理与生理学系

出　　处：《中华核医学杂志》2011年第2期82-86,共5页Chinese Journal of Nuclear Medicine

基　　金：国家自然科学基金（30500601,30872999）;江苏省自然科学基金（BK2007023） 志谢本研究还得到南京医科大学重点项目（06NMUZ044）及无锡市医院管理中心重大技术项目（ZD0604）资助

摘　　要：目的 观察纳米磁粒磁感应加热、单纯疱疹病毒-胸苷激酶（HSV-tk）基因及核素内照射联合治疗MCF-7乳腺癌的作用,探讨联合治疗的有效性与可行性.方法 制备转染试剂脂质体（Lipofectamine）/含热休克蛋白启动子的HSV-tk基因表达质粒（pHSP-HSV-tk）、四氧化三铁纳米磁流液、188Re-丙氧鸟苷白蛋白纳米微球（GCV-BSA-NP）;进行四氧化三铁纳米磁流液的体内加热实验;建立MCF-7乳腺癌模型,并用单纯随机抽样法将60只荷瘤鼠分成6组,每组10只.A组为空白对照组,B组为单纯基因转染治疗组,C组为单纯热疗组,D组为基因转染联合核素内照射治疗组,E组为基因转染联合热疗组,F组为基因、热疗、核素内照射联合治疗组.各组经不同方法处理后,观察肿瘤生长情况,测肿瘤质量,观察肿瘤组织病理变化;RT-PCR法检测B、D、E、F组HSV-tk基因表达（与内参β-actin mRNA吸光度比值）情况.各组与对照组比较采用泊松分布及单因素方差分析,所用统计软件为SPSS 10.0.结果 在动物体内加热实验中,交变磁场（AMF）作用下,不同注射剂量磁粒都引起肿瘤组织温度快速升高,2,4和6 mg磁粒分别使肿瘤温度升至39.6 ℃,43.2 ℃和48.1 ℃,且持续40 min.当AMF作用停止后,肿瘤组织温度10 min内分别降至36.8 ℃,37.5 ℃和37.8 ℃.成功建立MCF-7乳腺癌模型,治疗结果显示：C、D、E、F组肿瘤质量[（452.50±30.29）、（240.98±35.32）、（231.87±27.41）、（141.55±23.78）mg]较A组[（719.12±22.65）mg]差异有统计学意义（F=800.007,P＜0.01）,且以F组疗效最好;C组与E组肿瘤质量大于F组,差异有统计学意义（t=25.533,7.872,P均＜0.05）;D组较B组肿瘤质量明显减小,且差异有统计学意义（t=32.805,P＜0.05）.RT-PCR半定量分析显示：B、D组的表达量（0.33±1.30,0.46±0.12）与E、F组（0.66±0.13,0.74±0.11）相比差异有统计学意义（F=21.573,P＜0.05）.结论 热疗、基因治疗联合核Objective To investigate the antitumor therapeutic effect of combined therapy of magnetic induction heating by nano-magnetic particles, herpes simplex virus thymidine kinase gene（HSV-tk suicide gene） and internal radiation in mice bearing MCF-7 breast carcinoma. Methods The transfection reagents, plasmids heat shock protein-HSV-tk （pHSP-HSV-tk）, ferroso-ferric oxide nano-magnetic fluid flow and 188Re-ganciclovir-bovine serum albumin-nanopaticles （GCV-BSA-NP） were prepared. The heating experiments in vivo were carried out using ferroso-ferric oxide nano-magnetic fluid flow. Sixty mice tumor models bearing MCF-7 breast carcinoma were established and randomly divided into six groups. Group A was the control group, B was gene transfection therapy group, C was hyperthermia group, D was gene transfection therapy combined with radionuclide brachytherapy group, E was gene therapy combined with hyperthermia group, and F was gene therapy, hyperthermia combined with radionuclide brachytherapy group. The tumor growth, tumor mass and histopathological changes were evaluated. The expression of HSV-tk in the groups of B, D, E and F was detected by RT-PCR. Poisson distribution and one-way analysis of variance （ANOVA） were used for statistical analysis by SPSS 10.0 software. Results In the animal heating experiments, the temperature of tumor increased up to 39.6 ℃, 43.2 ℃, and 48.1 ℃ quickly with different injected doses （2, 4 and 6 mg respectively） of nano-magnetic particles and maintained for 40 min. The temperature of tumor tissue reduced to 36.8 ℃, 37.5 ℃ and 37.8 ℃ in 10 min when alternating magnetic field （AMF） stopped. The tumor mass in Groups C （（452.50 ±30.29） mg）, D （（240.98 ±35.32）mg）, E（（231.87 ±27.41） mg） and F （（141.55 ±23.78） mg） were much lower than that in Group A （（719.12±22.65） mg） （F=800.07, P〈0. 01）, with the most significant treatment effect in Group F.The tumor mass in Group B（（684.05 ±24.02） mg） was higher than

关 键 词：乳腺肿瘤 肿瘤细胞 培养的 磁性纳米颗粒 基因疗法 近距离放射疗法 小鼠 裸 

分 类 号：R737.9[医药卫生—肿瘤]