携带MIA基因单核细胞增生性李斯特菌抑制恶性黑素瘤的研究  

Inhibitory effect of Listeria monocytogenes expressing melanoma-inhibiting activity gene on malignant melanoma

作　　者：钱悦[1] 蒋苹[1] 褚淑娟[2] 陈思远[1] 冯爱平[1] 罗勤[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院皮肤科,武汉430022 [2]华中科技大学同济医学院附属协和医院麻醉科,武汉430022 [3]华中师范大学生命科学院

出　　处：《中华皮肤科杂志》2009年第6期399-401,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30600535）

摘　　要：目的探讨携带黑素瘤抑制性活性蛋白（MIA）基因的单核细胞增生性李斯特菌（LM）对黑素瘤的抑制作用。方法构建载体pERL3-hly—MIA，电转化单核细胞增生性李斯特菌建立携带MIA基因的单核细胞增生性李斯特菌（LM—hly—MIA），Western鉴定。实验小鼠随机分成生理盐水组（A组）、LM组（B组）、LM—hly—MIA组（C组），免疫后次日给予小鼠背部皮下接种1×10^7／mLB16细胞0．1mL。记录注射部位肿瘤的大小与小鼠的一般情况。结果经酶切及Western Blot鉴定表明成功构建LM—hly—MIA，体内试验显示生理盐水组、LM组、LM—hly—MIA组小鼠肿瘤平均重量分别为4．33±0．91g、3．36±0．41g、1．89±0．52g，其中LM的肿瘤抑制率为22．4％，LM—hly—MIA的肿瘤抑制率为61．5％（P〈0．05）。结论携带MIA基因减毒单核细胞增生性李斯特菌明显抑制黑素瘤B16的生长。Objective To investigate the inhibitory effects ofL. monocytogenes expressing melanomainhibiting activity （MIA） gene on malignant melanoma. Methods The plasmid pERL3-hly-MIA was con- structed and used to transform live L. monocytogenes by electroporation. After culture, the transformed L. monocytogenes, namely LM-hly-MIA, was harvested for the detection of MIA protein by Western Blot. A total of 24 mice were divided into three groups, namely control group, LM group and LM-hly-MIA group, to be immunized with physiological saline, L. monocytogenes and LM-hly-MIA, respectively. One day after the immunization, mice were subcutaneously inoculated with 0.1 mL of B16 cells at a concentra- tion of 1 × 10^7/mL. One week later, reimmunization was performed. The size and weight of tumors were observed and measured in these mice. Results Enzyme digestion and Westem blot confirmed the successful construction of plasmid pERL3-hly-MIA. The average weight of tumors in mice was 4.33 ± 0.91 gram, 3.36 ± 0.41 gram and 1.89± 0.52 gram, respectively in the control group, LM group and LM-hly-MIA group, respectively （P 〈 0.05 ）, with the inhibition rates of tumor growth being 22.4% and 61.5% in LM group and LM-hly-MIA group, respectively. Conclusion The attenuated strain of L. monocytogenes expressing MIA gene could evidently inhibit melanoma growth.

关 键 词：黑色素瘤 利斯特菌单核细胞 增生 抑制蛋白 模型 动物 

分 类 号：R739.5[医药卫生—肿瘤]