聚乙烯吡咯烷酮介导的白细胞介素-12基因治疗小鼠肝癌微小病灶的实验研究

Experimental study on interleukin-12 gene therapy induced by polyvinyl pyrrolidone for micro hepatoma in mouse

目的 探讨白细胞介素 (IL) 12基因注射对小鼠肝癌微小病灶的治疗作用。方法 IL 12质粒转染BHK 2 1细胞 ,ELISA检测上清中IL 12的分泌 ,并用此上清刺激脾细胞 ,MTT检测其增殖。18只Balb/c小鼠股部肌肉内接种H2 2细胞后分为 3组分别注射聚乙烯吡咯烷酮 (PVP) /生理盐水(NS)、PVP/NS作溶剂的 pCDNA3.1质粒和IL 2质粒观察肿瘤的形成 ;ELISA检测血清中IL 12的含量 ,切片观察肿瘤浸润淋巴细胞 ,并用免疫组化和TUNEL显示肿瘤血管密度和肿瘤细胞的凋亡。结果 IL 12因子可以分泌到胞外 ,促进脾淋巴细胞的增殖 (15 .2 % )。与对照组相比 ,治疗组血清IL 12水平上升 ,瘤内淋巴细胞大量浸润 ,血管减少 ,肿瘤细胞凋亡 ,肝癌微小病灶生长受抑 ,抑制率高达 85 .7%。结论 PVP介导的IL 12基因重复注射对小鼠肝癌微小病灶具有很好的治疗效果。

Objective To study the therapeutic effect of interleukin(IL)-12 gene therapy on micro hepatoma in mouse. Methods IL-12 in the supernate of BHK-21 cells transfected with pmIL-12 was assayed by ELISA and used to stimulate spleen lymphocytes,and its proliferation was evaluated by MTT. Eighteen Balb/c mice were randomly divided into three groups after inoculating hepatoma cells H22 in right leg muscle. The mice in different groups were injected with solutions of polyvinyl pyrrolidone(PVP)/NS,pcDNA3.1 or pmIL-12,every 3 days for 7 times. Tumor weight and serum IL-12 were assayed. Tumor infiltrating lymphocytes,tumor microvessel density and apoptosis were also examined by H-E staining,immumohistochemistry and TUNEL. Results Lymphocytes proliferation of spleen can be promoted to 15.2% by extracellular secreting of IL-12. Compared with control groups,tumor growth in gene therapy group was highly suppressed (85.7%) with higher level of serum IL-12,more infiltrating lymphocytes,fewer tumor vessels and more apoptosis cells. Conclusion IL-12 gene induced by PVP had its therapeutic efficacy on micro hepatoma in mouse.