突变Ki-ras基因修饰的肺癌DC疫苗的体外抗癌活性  

Anti-tumor activities of lung cancer dendritic cell vaccine modified by mutant Ki-ras gene in vitro

作　　者：喻璟瑞[1] 陆燕蓉[1] 朱文[1] 王艳萍[1] 陈小禾[1] 易成[1] 罗德云[1] Michael T.Lotze 周清华[1] 

机构地区：[1]四川大学华西医院四川省肺癌分子重点实验室,肿瘤中心,成都610041 [2]匹兹堡大学肿瘤中心肿瘤外科,美国宾夕法尼亚州15260

出　　处：《中国肺癌杂志》2004年第2期104-107,共4页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金 (No .30 0 0 0 2 0 6);高等学校博士点基金(No .2 0 0 0 55)资助~~

摘　　要：目的　探讨以Ki ras基因 12位密码子点突变为靶点 ,构建DC Ad Ki ras(V12 )肺癌疫苗及其抗肺癌免疫治疗的可能性。方法　应用重组腺病毒的方法将人Ki ras基因 12位密码子点突变 (由甘氨酸转变为缬氨酸 )的cDNA导入DC ,构建DC Ad Ki ras(V 12 )肺癌疫苗 ,以流式细胞术、PCR、MLR和CTL等方法检测其免疫活性。结果　 ( 1)DC Ad Ki ras(V12 )肺癌疫苗不仅能表达人Ki ras(V12 )基因 ,且能明显刺激T细胞增殖及提高CTL的杀伤作用。 ( 2 )突变Ki ras基因修饰的DC免疫小鼠后 ,可产生针对表达Ki ras(V 12 )基因的Lewis肺癌的特异性杀伤作用 ,但对B16黑色素瘤细胞则无明显杀伤作用。结论　以人突变Ki ras基因修饰的肺癌DC疫苗可在体外特异性诱导抗可表达Ki ras(V 12 )基因的Lewis肺癌的活性。Objective To construct a DC-Ad-Ki-ras(V12) vaccine and investigate the anti-tumor activities of lung cancer dendritic cell vaccine modified by mutant Ki-ras gene in vitro. Methods Ki-ras(V12) cDNA was transfected into cultured bone marrow-derived DC with the recombinant adenovirus [(Ad-Ki-ras(V12)] containing human mutant Ki-ras gene. Anti-tumor activity of the vaccine was studied in vitro by flow cytometry, PCR, MLR and cytotoxicity assay. Results (1) The DC vaccine was confirmed not only to express Ki-ras(V12) gene, but also to remarkably stimulate lymphocyte proliferation and improve CTL activity. (2) The DC vaccine modified by mutant Ki-ras gene could induce specifical CTL activity of immunized mice against Lewis lung carcinoma that could express Ki-ras(V12) gene, but not to B16. Conclusion The DC vaccine modified by mutant Ki-ras gene can induce obvious anti-tumor activities against Lewis lung carcinoma that can express Ki-ras(V12) gene.

关 键 词：基因突变 KI-RAS基因 基因修饰 肺癌 DC 疫苗 抗癌活性 树突状细胞 

分 类 号：R734.2[医药卫生—肿瘤] R394[医药卫生—临床医学]