可诱导表达的白细胞介素12基因治疗小鼠原位移植性肝癌  

Gene therapy based on inducible IL-12 gene in a mouse model of orthotopically transplanted H22 hepatoma

作　　者：李艳茹[1] 张海英[1] 金珍婧[2] 李玉林[1] 王琳[1] 

机构地区：[1]吉林大学白求恩医学院病理生物学教育部重点实验室,长春130021 [2]吉林大学第二医院消化内科

出　　处：《中华肿瘤杂志》2010年第7期487-491,共5页Chinese Journal of Oncology

基　　金：教育部新世纪优秀人才支持计划（NCET-04-0303）;国家自然科学基金（30370547）;教育部优秀青年教师资助计划（2003）

摘　　要：目的 评估RU486系统诱导表达的白细胞介素12（IL-12）基因对小鼠原位移植性肝癌的治疗效果.方法 小鼠肝脏接种H22肝癌细胞,制备原位移植性肝癌模型.采用水流动力学注射法,将含有RU486调控系统的小鼠IL-12表达质粒pRS22注射到小鼠体内,腹腔注射RU486（250μg/kg）诱导质粒表达,检测血清中IL-12、干扰素γ（IFN-γ）和一氧化氮（NO）的表达水平.采用HE染色和免疫组织化学方法检测瘤细胞增殖活性和肿瘤血管生成情况.结果 注射生理盐水＋RU486组、pRS-LacZ＋RU486组、pRS22＋芝麻油组和pRS22＋RU486组小鼠肝脏肿瘤体积分别为（409.90±137.03）mm^3、（271.80±182.63）mm^3、（251.00±76.55）mm^3和（46.48±47.19）mm^3,肿瘤细胞增殖细胞核抗原（PCNA）阳性率分别为（82.10±4.62）%、（83.45±2.34）%、（77.46±2.99）%和（50.67±8.09）%,肿瘤组织微血管密度分别为（74.58±18.47）个/400倍视野、（63.60±13.36）个/400倍视野、（53.52±11.74）个/400倍视野和（25.38±10.87）个/400倍视野.与注射生理盐水＋RU486组、pRS-LacZ＋RU486组和pRS22＋芝麻油组相比,pRS22＋RU486组小鼠生存期明显延长（接种瘤细胞后第50天仍然有1只小鼠存活）,血清中IL-12、IFN-γ和NO的表达水平分别为（11.52±7.46）ng/ml、（2.68±0.32）ng/ml和（5.10±3.27）μmol/L.结论 RU486系统能有效地调控IL-12基因的表达,可诱导的IL-12能够有效地抑制小鼠原位移植性肝癌的生长,延长小鼠的生存期.Objective To evaluate the antitumor efficiency of IL-12 gene induced by RU486 regulatory system in a mouse model of orthotopically transplanted hepatoma. Methods The orthotopic hepatoma model was prepared by inoculation of H22 hepatoma cells into the mouse liver. Murine interleukin-12 （IL-12） expressing plasmid pRS22 containing RU486 regulatory system was injected into mice by a hydrodynamic injection 3 days after H22 cells inoculation. Three days after hydrodynamic injection, the mice were induced with RU486 （250 μg/kg） consecutive intraperitoneal administration for 6 days. Blood samples were taken at 10 h after the first and third induction for the determination of IL-12, IFN-γ and NO. Five mice were sacrificed at 2 days after the treatment with RU486. The tumor size was measured. HE and immunohistochemical stainings were applied to evaluate the proliferative activity and angiogenesis in the tumors. The other 7 mice were kept to monitor their survival. Results In mice receiving saline plus RU486, pRS-LacZ plus RU486, or pRS22 plus sesame oil, the liver tumors were big in size; （409. 90± 137.03） mm^3, （271. 80±182.63） mm^3 and （251.00±76. 55） mm^3, respectively. Strong PCNA positive expression [ （82.10±4. 62） % , （83. 45±2. 34）% and （77. 46±2. 99）% ] and extensive microvessel density （74.58±18.47, 63.60±13. 36 and 53.52±11.74 per 400×field）, respectively, in these tumor tissues were observed after immunohistochemical staining. The survival period was shorter in these mice. In contrast, in mice treated with pRS22 plus RU486, the tumor was smaller in size. Extensive necrosis, weak PCNA proliferative activity （50.67±8. 09）% , and a marked paucity of microvessel density （25. 38±10. 87） were seen. The survival of mice was obviously prolonged. Compared with the 3 control groups, a significant elevation of serum IL-12, IFN--γ and NO levels were detected in the mice treated with pRS22 plus RU486. Conclusion Expression of IL-12 gene can be effectively controlled

关 键 词：肝肿瘤 白细胞介素12 RU486 基因疗法 小鼠 

分 类 号：R735.7[医药卫生—肿瘤]