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(P<0.05);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(P<0.05)。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(P>0.05)。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。
By observing JSRV Env eukaryotic expression plasmid transfected mouse NIH 3T3 （mouse fibroblast cell line） on influence of vascular endothelial growth factor （VEGF）and epider- mal growth factor receptor （EGFR） expression to further explore a relationghip between their changes and sheep pulmonary adenomatosis （OPA） association tumorigenesis. First in vitro cul- tured NIH3T3 cell to establish transfected pEGFP-CI-env, pEGFP-C1 and non-transfected cells. Then used real-time quantitative PCR method to detect the expression of VEGF and EGFR mRNA level and ELISA method to detect the expression of two cell factors in NIH 3T3 cell culture super- natant with different treatments. Compared with pEGFP-C1 and non-transfected cells,VEGF and EGFR mRNA expression in transfected with pEGFP-CI-env cell group was significantly increased （P〈0.05） ,and the VEGF and EGFR protein concentration in the cell culture supernatant was sig- nificantly increased（P〈0.05）. Compared to non-transfected cells group VEGF and EGFR mRNA and protein expression levels had no significant difference in cells transfected with pEGFP-C1 group （P〉0.0;）. In transfected NIH 3T3 cell transformation process in mice induced by JSRV Env VEGF and EGFR expression levels were increased , speculating that it may play a key role in OPA tumor. It provide a theoretical basis for gene therapy targeting based on OPA.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2017年第5期898-904,共7页
Chinese Journal of Veterinary Science
