mTERT和mTyr双启动子联合调控HN基因重组腺病毒对B16动物模型的抑瘤作用  

Anti-tumor effects of recombinant adenovirus with HN genes co-regulated by mTERT and mTyr promoters on B16 nude mice model

作　　者：孟佳丽 龙雨清 林静 李昕 尚翠玲 王小月 蓝肇煜 李丹 金天明 MENG Jia-li;LONG Yu-qing;LIN Jing;LI Xin;SHANG Cui-ling;WANG Xiao-yue;LAN Zhao-yu;LI Dan;JIN Tian-ming(College of Animal Science and Veterinary Medicine , Tianjin Agricultural University , Tianjin 300384,China;College of Life Science , Nankai University ,Tianjin 300071, China;College of Veterinary Medicine ,Jilin University,Changchun 130062 ,China)

机构地区：[1]天津农学院动物科学与动物医学学院,天津300384 [2]南开大学生命科学院,天津300071 [3]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2019年第10期1921-1926,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31572492);天津市科技计划高端兽药先进制造科技重大专项基金资助项目(17ZXGSNC00030);天津市农委农业科技示范推广资助项目(201601380)

摘　　要：为探讨双启动子调控HN基因重组腺病毒Ad-mTERTp-mTyrp-HN对小鼠黑色素瘤动物模型的抑瘤作用,试验构建B16裸鼠皮下移植瘤模型,采用HEK293细胞扩增重组腺病毒,进行纯化和滴度检测。将重组腺病毒Ad-HN、Ad-mTERTp-HN、Ad-mTyrp-HN、Ad-mTERTp-mTyrp-HN、Ad-GFP和PBS通过尾静脉注射裸鼠,冰冻切片观察裸鼠肿瘤内GFP绿色荧光表达情况,通过肿瘤生长和裸鼠存活情况计算抑瘤率,探究其在体内的抗肿瘤作用。结果显示:重组腺病毒在HEK293细胞中扩增,滴度分别为10^11.250,10^12.250,10^10.625,10^12.125,10^12.250 TCID50/mL;肿瘤组织中能观察到黑色素细胞,证明皮下黑色素移植瘤模型构建成功;重组腺病毒对脏器无损伤,且在双启动子组脏器中没有观察到黑色素细胞的肝肺转移;通过尾静脉注射重组腺病毒后能够在肿瘤中观察到荧光表达;与其他组相比双启动子组重组腺病毒不但能抑制黑色素瘤的生长,而且能延长裸鼠的生存期,抑瘤率为55.5%。因此,该重组腺病毒Ad-mTERTp-mTyrp-HN具有靶向性,并且可以抑制肿瘤生长,可为黑色素瘤的治疗提供参考。To investigate anti-tumor effects of dual promoters and HN gene recombinant adenovirus Ad-m TERTp-m Tyrp-HN on melanoma nude mice model,a subcutaneous xenograft model of B16 nude mice was constructed and recombinant adenovirus was amplified by HEK293 cell for purifi cation and titer determination.The recombinant adenovirus Ad-H N, Ad-mTERTp-H N, Ad-mTyrp- H N,Ad-m TERTp-m Tyrp-HN and Ad-GFP were injected through the tail vein of nude mice,and GFP green fluorescence in nude mice was observed by frozen section. The tumor growth and sur vival of nude mice were recorded,and tumor inhibition rate was calculated to explore anti-tumor effect in vivo . Recombinant adenovirus was amplified in HEK293 cells, with titers of 10^11.250, 10^12.250, 10^10.625 , 10^12.125 and 10^12.250 TCID50/m L, respectively. Melanin can be observed in tumor tissue, which proved that the subcutaneous tumor model of melanoma in nude mice was successfully constructed.Compared with other groups, recombinant adenovirus in double-promoter group not only inhibited the growth of melanoma,but also prolonged the survival of nude mice with the inhibition rate of 55.5 Therefore, this recombinant adenovirus Ad-mTERTp-mTyrp-HN has the targeted character which can inhibit tumor grow th, providing a reference for the treatment of melanoma.

关 键 词：重组腺病毒 HN基因 mTERT启动子 mTyr启动子 B16裸鼠 黑色素瘤 

分 类 号：R540[医药卫生—心血管疾病] S852.61[医药卫生—内科学]