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目的:探讨T-bet质粒局部应用对小鼠肺癌细胞皮下移植瘤的干预作用。方法:将传代培养的Lewis肺癌细胞接种于小鼠右肩部皮下,制备荷瘤小鼠模型;瘤内分别注射PIRES-eGFP/mT-bet(m指小鼠)、PIRES-eGFP质粒和PBS,观察各组小鼠存活时间,肿瘤体积的变化;HE染色观察肿瘤组织的病理变化;Western blot检测肿瘤组织T-bet的表达,QRT-PCR检测肿瘤组织T-bet和IFN-γ的mRNA水平。结果:瘤内注射T-bet质粒载体后Western blot检测发现,肿瘤组织T-bet蛋白表达显著高于对照组;QRT-PCR结果显示,肿瘤组织T-bet和IFN-γ的mRNA水平呈上升趋势,与对照组相比差异有统计学意义(P<0.05)。T-bet质粒干预组小鼠肿瘤生长明显缓慢,存活期延长。HE染色组织病理观察可见mT-bet干预组小鼠局部组织淋巴细胞大量浸润。结论:T-bet基因局部注射可延缓小鼠肺癌Lewis皮下移植瘤的生长,提高机体的抗肿瘤免疫应答能力,增强对已发肿瘤的生长限制作用,为T-bet应用于肿瘤治疗积累试验数据。
Objective: To investigate the effect of local application of T-bet plasmid on subcutaneous xenografts of lung cancer cells in mice. Methods: Lewis lung carcinoma cells were subcutaneously inoculated on the right shoulder of mice to establish tumor-bearing mice model. PIRES-eGFP / mT-bet (m mice), PIRES- The survival time and tumor volume of each group were observed. The pathological changes of the tumor tissue were observed by HE staining. The expression of T-bet in the tumor tissue was detected by Western blot. The mRNA levels of T-bet and IFN-γ in the tumor tissue were detected by QRT-PCR. Results: The expression of T-bet protein in tumor tissue was significantly higher than that in control group by Western blot after the T-bet plasmid vector was injected intratumorally. The results of QRT-PCR showed that the mRNA level of T-bet and IFN- , Compared with the control group, the difference was statistically significant (P <0.05). T-bet plasmid intervention mice tumor growth was slow, prolonged survival. Histopathological examination showed that the mT-bet intervention group, a large number of local tissue lymphocytes infiltration. CONCLUSION: Local injection of T-bet gene can delay the growth of Lewis lung carcinoma in mice with lung cancer, enhance the anti-tumor immune response ability of the mice and enhance the growth restriction of the established tumor, and accumulate the experimental data for the application of T-bet in tumor therapy .