纳米磁性干扰素脂质体在裸鼠移植性人肝癌靶向治疗过程中对VEGF和Caspase-3表达的影响

目的研究纳米磁性重组人干扰素-α2b脂质体在裸鼠移植性人肝癌靶向治疗过程中对移植瘤模型中瘤组织VEGF和Caspase-3表达的影响。方法30只荷瘤裸鼠随机分成5组,每组6只,尾静脉注射给药〔生理盐水组,NS组:0IUIFN.(200μl)-1;游离干扰素组,IFN组:10000IUIFN.(200μl)-1;脂质体干扰素组,IL组:1000IUIFN.(200μl)-1;磁性空白脂质体组,ML组:0IUIFN.(200μl)-1;磁性干扰素脂质体组,MIL组:1000IUIFN.(200μl)-1〕,在肿瘤部位施加30min外磁场(5000GS),共进行3次,治疗后d30处死动物,测瘤体积、称瘤重并计算抑制率。采用RT-PCR法检测各组肿瘤组织VEGF和Caspase-3mRNA表达,Westernblot法检测各组肿瘤组织VEGF和Caspase-3蛋白表达。结果MIL对人肝癌移植瘤有明显抑制作用,抑瘤率为62.50%,高于IFN(27.61%)和IL(28.17%);RT-PCR显示MIL的VEGF mRNA表达明显低于对照组和其它实验组(P<0.01),而MIL的Caspase-3mRNA表达明显高于对照组和其它实验组(P<0.01);Western blot显示MIL的VEGF蛋白表达明显低于对照组和其它实验组(P<0.01),而MIL的Caspase-3蛋白表达明显高于对照组和其它实验组(P<0.01)。结论在外加磁场作用下,MIL在体内有明显的靶向治疗肿瘤效果:通过下调VEGF、上调Caspase-3在mRNA和蛋白方面的表达,能够抑制肿瘤血管增生和促进肿瘤细胞凋亡,从而进一步抑制肝癌的生长与转移。

Effect of nanometer magnetic interferon-α2b liposome on expression of VEGF and Caspase-3 in the process of targeting therapy Human Hepatocelluar carcinoma Implants of Nude Mice

Aim To evaluate the effect and mechanism of vascular endothelial growth factor(VEGF) and Cysteine protease aspartic acid-3(Caspase-3) inhibition of human hepatocellular carcinoma(HCC)implants in nude mice by nanometer magnetic interferon-α2b liposome targeting therapy.Methods Thirty nude mices bearing human HCC were divided into 5 groups of 6 mice:group 1,saline(NS);group 2,free interferon-α2b group(IFN);group 3,interfe-ron-α2b liposome group(IL);group 4,magnetic liposome group(ML);group 5,nanometer magnetic interferon-α2b liposome group(MIL).Above dosage of 200 μl/nude mice was respectively injected through the caudal vein in these 6 groups.Meantime,magnetic field with the strength of 5000 GS was added to the tumor surface about 30 min,and this therapy was done three times.After 30 days,the mices were killed and the tumors were taken out.The difference at the mRNA expression level of VEGF and Caspase-3 was observed by RT-PCR between the experimental group and control group.The protein expression level of VEGF and Caspase-3 was detected by western blot between the experimental group and control group.Results Targeting therapy study in nude mice bearing human HCC displayed that the growth speed of tumor in the MIL group significantly slowed down than other groups.The tumor inhibition rate of MIL group was 62.50%,which was remarkably higher than those of the IFN group(27.61%) and IL group(28.17%).RT-PCR showed that the mRNA expression of VEGF of MIL group is the lowest of all groups(P<0.01) and the mRNA expression of Caspase-3 of MIL group is the highest of all groups(P<0.01).Western blot showed that the VEGF165 protein(monomer,P21) expression is the lowest of all groups(P<0.01)and the cleaved Caspase-3 protein(p17) expression is the highest of all groups(P<0.01).Conclusion The MIL has an obvious targeting therapeutic effect in vivo under a magnetic field.By downregulating the mRNA and protein expression of VEGF and upregulating the mRNA and protein expression Caspase-3,the MIL may suppress the angiogensis and promote the apoptosis of tumor cell and further inhibit growth and metastasis of hepatocelluar carcinoma.