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目的:水泡口炎病毒(Vesicular Stomatitis Virus,VSV)基质蛋白(Matrix protein,M蛋白)具有诱导肿瘤细胞凋亡的作用,本研究探讨水泡口炎病毒基质蛋白对癌性腹水形成的抑制和治疗作用。方法:采用旋转蒸发仪法制备纳米脂质体,检测其体外转染效率;采用脂质体转染技术将已构建的水泡口炎病毒基质蛋白(VSV-M)重组真核表达质粒pcDNA3.1-M转入MethA肿瘤细胞,转染后6小时将细胞接种于小鼠腹腔,观察小鼠腹水的形成情况;腹水治疗组,则先将MethA肿瘤细胞接种于小鼠腹腔,将小鼠随机分成4组,于接种后第二天分别用脂质体包裹的pcD-NA3.1-M、pcDNA3.1空载体、单纯脂质体及生理盐水治疗,观察腹水的形成情况。结果:自制的DOTAP:DOPE脂质体与Sigma公司购买的Lipofectamine 2000的转染效率相似,pcDNA3.1-M明显抑制MethA腹水的形成,对已经形成的腹水也有明显的治疗作用,与对照组比较有统计学意义(P<0.05),同时明显延长了小鼠的存活期。结论:VSV-M蛋白真核表达质粒pcDNA3.1-M对小鼠腹水的形成有抑制作用,能延长小鼠的存活期,对于恶性腹水的治疗具有一定的意义,值得进一步研究。
OBJECTIVE: Vesicular stomatitis virus (VSV) matrix protein (M protein) can induce the apoptosis of tumor cells. This study was to investigate the inhibitory and therapeutic effects of vesicular stomatitis virus matrix protein on the formation of cancerous ascites . Methods: Nano-liposomes were prepared by rotary evaporation method and their transfection efficiency was tested in vitro. The constructed recombinant plasmid pcDNA3.1 of vesicular stomatitis virus matrix protein (VSV-M) -M into MethA tumor cells, 6 hours after transfection, the cells were inoculated into the abdominal cavity of mice to observe the formation of ascites in mice; ascites group, the first MethA tumor cells were inoculated into the abdominal cavity of mice, the mice were randomly divided into 4 groups. The pcD-NA3.1-M and pcDNA3.1 empty vector, which were encapsulated by liposome, were treated with liposome and saline respectively on the second day after inoculation to observe the formation of ascites. Results: The transfection efficiency of homemade DOTAP: DOPE liposomes was similar to that of Lipofectamine 2000 purchased from Sigma. PcDNA3.1-M significantly inhibited the formation of ascites of MethA and also had a significant therapeutic effect on the ascites that had formed. Compared with the control group There was statistical significance (P <0.05), while significantly prolonging the survival of mice. CONCLUSION: The eukaryotic expression plasmid pcDNA3.1-M of VSV-M inhibits the formation of ascites in mice and prolongs the survival of mice. It is of great significance for the treatment of malignant ascites, which deserves further study.