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[目的]研究重组质粒pEGFP-N1-白细胞介素(IL)-24基因及pGEG-IL-12基因单独应用与联合应用对小鼠黑色素瘤移植瘤的抑制作用及其机制.[方法]制备黑色素瘤B16细胞荷瘤小鼠模型,并随机分为5个组,分别向瘤体内注射给予脂质体包裹的基因重组质粒pEGFP-N1-IL-24(IL-24组),pEGFP-N1-IL-24和pGEG-IL-12(联合应用组),pGEG-IL-12(IL-12组),pEGFP-N1(空质粒组)及PBS(PBS组).每次给药前测量小鼠背部肿瘤的长径和短径,绘制肿瘤生长曲线.采用HE染色法观察各组肿瘤组织形态;RT-PCR法检测移植瘤组织中IL-24,IL-12,Bax,Bcl-2,VEGF,caspase-3及caspase-12mRNA的表达;Western Blot检测移植瘤组织中Bax,Bcl-2,VEGF,caspase-3及caspase-12蛋白的表达.[结果]肿瘤生长曲线观察结果表明,IL-24组、IL-12组及联合应用组肿瘤体积均明显小于空质粒组和PBS组(P<0.05),联合应用组肿瘤体积最小(P<0.05).HE染色结果显示,空质粒组多数细胞生长差,形态不规则,细胞皱缩,结构稀疏,细胞核溶解、变形,细胞成片坏死.RT-PCR检测结果表明,IL-24组和IL-12组分别有IL-24和IL-12基因mRNA表达,联合应用组同时有IL-24和IL-12基因mRNA表达,而空质粒组和PBS组均未见表达;IL-24组、IL-12组及联合应用组Bax基因mRNA表达水平均明显高于空质粒组和PBS组(P<0.05),联合应用组Bax基因mRNA的表达水平最高,而Bcl-2和VEGF基因mRNA的表达水平则显著降低(P<0.05).Western Blot检测结果表明,IL-24组、IL-12组及联合应用组Bax,caspase-3,caspase-12蛋白的表达水平均明显高于空质粒组和PBS组(P<0.05),联合应用组中Bax,caspase-3,caspase-12蛋白的表达水平最高,而Bcl-2和VEGF蛋白的表达水平则显著降低(P<0.05).[结论]IL-24和IL-12对荷瘤小鼠有治疗作用,二者联合应用治疗效果更强.
[Objective] To investigate the inhibitory effect of recombinant plasmid pEGFP-N1-IL-24 and pGEG-IL-12 gene on mouse melanoma xenografts and its mechanism. [Methods] The tumor-bearing mice model of B16 tumor cells were randomly divided into five groups. The recombinant plasmid pEGFP-N1-IL-24 (IL-24 group) and pEGFP-N1-IL -24 and pGEG-IL-12 (combination group), pGEG-IL-12 (IL-12 group), pEGFP-N1 (empty plasmid group) and PBS Tumor growth curves were drawn according to the long and short diameters of the tumor, and the morphological changes of the tumors were observed by HE staining. The expressions of IL-24, IL-12, Bax, Bcl-2, VEGF and caspase The expression of Bax, Bcl-2, VEGF, caspase-3 and caspase-12 in the tumor tissue were detected by Western Blot. [Results] The observation of tumor growth curve showed that IL-24, The volume of tumor in IL-12 group and combination group was significantly smaller than that in empty plasmid group and PBS group (P <0.05), and the combined application group had the smallest tumor volume (P <0.05) .HE staining results showed that most cells in empty plasmid group grew poorly, Form is not The results of RT-PCR showed that the expression of IL-24 and IL-12 mRNA was detected in IL-24 and IL-12 groups, respectively, and the combination of them The expression of IL-24 and IL-12 mRNA was also observed in both groups, but not in empty plasmid group and PBS group. The expression of Bax mRNA in IL-24 group, IL-12 group and combination group was significantly higher than that in empty plasmid Group and PBS group (P <0.05), the expression of Bax gene mRNA and the mRNA expression of Bcl-2 and VEGF were significantly decreased (P <0.05) .Western Blot results showed that IL-24 The levels of Bax, caspase-3 and caspase-12 in IL-12 group and combination group were significantly higher than those in empty plasmid group and PBS group (P <0.05). The levels of Bax, caspase-3, caspase The expression of Bcl-2 and VEGF protein were significantly decreased (P <0.05). [Conclusion] IL-24 and IL-12 have a therapeutic effect on tumor-bearing mice, and the combination of the two The treatment effect is stronger.