JSRV-Env诱导转染NIH 3T3细胞后对VEGF和EGFR因子表达的影响

发布时间：2018-07-13 21:08

【摘要】：通过探索JSRV Env真核表达质粒诱导转染小鼠NIH 3T3(小鼠成纤维细胞)后对血管内皮生长因子(VEGF)和表皮生长因子受体(EGFR)表达的影响,进一步探讨其变化与绵羊肺腺瘤病(OPA)肿瘤发生的关联性。首先体外培养NIH 3T3细胞,设立转染pEGFP-C1-env、pEGFP-C1及未转染细胞组。本研究采用实时荧光定量PCR方法检测VEGF和EGFR mRNA水平上的表达;ELISA方法检测两细胞因子在不同处理的NIH 3T3细胞培养上清中蛋白的表达。结果显示,相比pEGFP-C1和未转染细胞组,转染pEGFP-C1-env细胞组中VEGF和EGFR mRNA的表达量均显著升高(P0.05);且在该处理组的细胞培养上清中检测到VEGF和EGFR蛋白浓度呈明显上升趋势,差异均有统计学意义(P0.05)。转染pEGFP-C1细胞组相比未转染细胞组VEGF和EGFR mRNA和蛋白表达量均差异不明显(P0.05)。结果表明,JSRV Env在诱导转染小鼠NIH 3T3细胞转化过程中,通过qPCR和ELISA的检测结果均可得知VEGF和EGFR表达量均升高,由两者的过表达变化推测VEGF和EGFR在OPA肿瘤发生过程中可能起关键作用,同时为OPA针对两者进行的基因靶向治疗方法提供理论基础。
[Abstract]:To explore the effect of JSRV Env eukaryotic expression plasmid on the expression of vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) in mouse NIH 3T3 (mouse fibroblasts). To further investigate the relationship between the changes and the tumorigenesis of sheep pulmonary adenomatosis (OPA). First, NIH 3T3 cells were cultured in vitro, then pEGFP-C1-envc1 transfected pEGFP-C1 and untransfected cells were established. In this study, the expression of VEGF and EGFR mRNA in NIH 3T3 cells was detected by real-time fluorescence quantitative PCR and the expression of two cytokines in the supernatant of NIH 3T3 cells was detected by Elisa. The results showed that the expression of VEGF and EGFR mRNA in pEGFP-C1-env cells was significantly higher than that in pEGFP-C1 and untransfected cells (P0.05), and the concentration of VEGF and EGFR protein in the culture supernatant of the treated group was significantly increased. The difference was statistically significant (P0.05). The expression of VEGF and EGFR mRNA and protein in pEGFP-C1 cells group was not significantly different from that in untransfected cells group (P0.05). The results showed that the expression of VEGF and EGFR increased by qPCR and Elisa during the induction of NIH 3T3 cell transformation by JSRV Env. The overexpression of VEGF and EGFR may play a key role in the development of OPA tumor and provide a theoretical basis for the gene targeting therapy of OPA.
【作者单位】：内蒙古农业大学兽医学院;农业部动物疾病临床诊疗技术重点实验室;
【分类号】：S852.65

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