人源JAZF1基因真核表达载体的构建及其对胚胎癌细胞凋亡的影响  

Construction of Eukaryotic Expression Vector of Human Juxtaposed with Another Zincfinger Gene 1 and Its Influence on Embryonal Carcinoma Cell Apoptosis

作　　者：陈倩 李超[1] 杨诚诚 母彬 龚欢 黄超[1] CHEN Qian;LI Chao;YANG Chengcheng;MU Bin;GONG Huan;HUANG Chao(College of VeterinaryMedicine,Sichuan Agricultural University,Chengdu 611130,China)

机构地区：[1]四川农业大学动物医学院,实验动物疾病模型研究室,成都611130

出　　处：《四川农业大学学报》2021年第1期108-113,共6页Journal of Sichuan Agricultural University

基　　金：四川农业大学学科建设双支计划(2018-2019)。

摘　　要：【目的】构建人源转录因子锌指并列基因1(JAZF1)真核表达载体,并探究过表达JAZF1对胚胎癌细胞存活的影响。【方法】以胚胎癌细胞NCCIT的cDNA为模板扩增JAZF1基因片段,并将其与pRK5-myc载体连接,构建重组质粒,转染至NCCIT细胞;通过实时荧光定量PCR和DAPI染色法分别检测过表达JAZF1对NCCIT细胞中炎性因子IL1-β、IL8、TGF-β转录水平的影响及对NCCIT细胞存活的影响。【结果】过表达JAZF1基因后,促炎因子IL1-β(P <0.05)、IL8(P <0.01)转录水平明显下降,而抑炎因子TGF-β的mRNA表达水平无明显变化(P>0.05),同时NCCIT细胞出现更明显的细胞凋亡(P<0.01)。【结论】JAZF1可能通过抑制促炎因子IL1-β和IL8的表达来抑制炎症发展,进而抑制畸胎瘤进展。【Objective】The eukaryotic expression vector of human juxtaposed with another zincfinger gene 1(JAZF1)was constructedto explore the effect of JAZF1 on embryonal carcinoma cell survival.【Method】The JAZF1 gene fragment was amplified using the cDNA of embryonic cancer cell NCCIT as a template,and wascloned into pRK5-myc vector to construct a recombinant plasmid.The recombinant plasmid was transfected into NCCIT cells,while the cell viability and the expression of transcription factorIL1-β,IL8 and TGF-βwere following evaluated by DAPI staining and quantitative real-time PCR,respectively.【Result】Transfected with JAZF1,the transcriptional levels of proinflammatory factor IL1-βand IL8 in NCCIT cells showed a significant suppression,while enhanced expression of antiinflammatory factor T GF-βwas observed.Meanwhile,we found increased cellularapoptosis in JAZF1-transfected NCCIT cells.【Conclusion】JAZF1 may suppressinflammation by inhibiting the expression of IL1-β and IL8,and then inhibit the progressionofembryonic cancer.

关 键 词：锌指并列基因1 胚胎癌 炎性因子 细胞凋亡 

分 类 号：Q78[生物学—分子生物学]