摘要
目的:探讨以中期因子(midkine,MK)基因为启动子的条件复制型溶瘤腺病毒(conditionally replicating oncolytic adenoviruses)Ad-MK单独或联合丝裂霉素(mitomycin,MMC)对膀胱移行细胞癌EJ细胞增殖的影响及可能的作用机制。方法:采用RT-PCR法检测EJ细胞中MKmRNA及感染Ad-MK后腺病毒E1amRNA的表达情况。MTT法检测Ad-MK和MMC单独或联合作用对膀胱癌细胞存活率的影响;光学显微镜下观察二者单独作用对EJ细胞形态的影响。通过测定病毒复制量观察MMC对Ad-MK复制的影响。建立膀胱移行细胞癌EJ细胞小鼠移植瘤模型,并观察Ad-MK单独或联合MMC对小鼠移植瘤生长的影响。结果:EJ细胞中可见MKmRNA的表达,感染Ad-MK后EJ细胞中可检测到腺病毒E1amRNA的表达。Ad-MK对膀胱癌细胞的增殖抑制作用呈明显的时间及剂量依赖性。Ad-MK联合低剂量MMC对EJ细胞生长的抑制作用明显增加,二者联合有协同作用。病毒复制量的测定发现,低剂量MMC能增加Ad-MK病毒在细胞中的复制作用,与单独感染病毒Ad-MK组相比,差异具有统计学意义(P<0.05)。小鼠体内实验证明,Ad-MK联合MMC治疗组小鼠的肿瘤生长缓慢,与单独感染病毒组比较,差异有明显的统计学意义(P<0.01)。结论:Ad-MK可在膀胱移行癌细胞中复制,MMC与Ad-MK联合作用具有协同效应,能上调病毒在细胞中的复制能力,从而增强对膀胱移行癌细胞的抑制作用。
Objective： To investigate the effect of conditionally replicating oncolytic adenoviruses with promoter of MK （midkine） （Ad-MK） alone or in combination with MMC （mitomycin） on proliferation of human bladder carcinoma EJ cells and its possible mechanism. Methods： The expression levels of MK mRNA in EJ cells and adenovirus E1a mRNA in Ad-MK-infected EJ cells were detected by RT-PCR （reverse transcription-PCR）. The effect of Ad-MK alone or in combination with MMC on survival rate of EJ cells was detected by MTT method, and the morphological change of EJ cells was observed under an optical microscope. Virus yield of Ad-MK was determined by TCID50 （50% tissue culture infectious dose） in the presence of MMC. A murine xenograft model of human bladder transitional cell cancer was established. The effect of Ad-MK alone or in combination with MMC on the growth of the murine xenografts was examined. Results： The MK mRNA was expressd in the EJ cells. The expression of E1a mRNA could be detectable in the Ad-MK-infected EJ cells. Ad-MK inhibited the proliferation of the cells in a dose- and time-dependent manner. There was a synergistic effect of Ad-MK in combination with MMC. The EJ cells infected with Ad-MK and treated with MMC produced a larger amount of viruses than the EJ cells infected with Ad-MK and not treated with MMC did in vitro and in vivo （P 〈 0.05）. The growth of murine xenografts of Ad-MK-infected and MMC-treated EJ cells was inhibited as compared with that of the murine xenografts of Ad-MK-infected EJ cells （P 〈 0.01）. Conclusion： Ad-MK can selectively replicate and inhibit the proliferation of EJ cells in a dose- and time-dependent manner. There maybe a synergistic effect of Ad-MK in combination with MMC. The potential growth inhibition effects of combined treatment proves to be superior to either virus or MMC treatment alone, and MMC can augment virus replication.
出处
《肿瘤》
CAS
CSCD
北大核心
2013年第7期575-580,共6页
Tumor
关键词
膀胱肿瘤
腺病毒科
病毒复制
丝裂霉素
Urinary bladder neoplasms
Adenoviridae
Virus replication
Mitomycin
