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

doi:10.16036/j.issn.1000-2650.2021.01.016

四川农业大学学报 ›› 2021, Vol. 39 ›› Issue (1): 108-113.doi: 10.16036/j.issn.1000-2650.2021.01.016

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

陈倩, 李超, 杨诚诚, 母彬, 龚欢, 黄超^*

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

收稿日期:2020-07-25 出版日期:2021-02-28 发布日期:2021-03-05
作者简介:陈倩,博士研究生。*责任作者:黄超,博士,副教授,主要从事细胞生物学和实验动物疾病模型研究,E-mail:huangchao@sicau.edu.cn。
基金资助:
四川农业大学学科建设双支计划(2018-2019)

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

CHEN Qian, LI Chao, YANG Chengcheng, MU Bin, GONG Huan, HUANG Chao^*

College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China

Received:2020-07-25 Online:2021-02-28 Published:2021-03-05

摘要/Abstract

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。

关键词: 锌指并列基因1, 胚胎癌, 炎性因子, 细胞凋亡

Abstract: 【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-β were following evaluated by DAPI staining and quantitative real-time PCR,respectively.【Result】 Transfected with JAZF1,the transcriptional levels of proinflammatory factor IL1-β and IL8 in NCCIT cells showed a significant suppression,while enhanced expression of antiinflammatory factor TGF-β was 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.

Key words: human juxtaposed with another zinc finger gene 1 (JZAF1), embryonal carcinoma, inflammat-oryfactor, apoptosis

中图分类号:

陈倩, 李超, 杨诚诚, 母彬, 龚欢, 黄超. 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[J]. 四川农业大学学报, 2021, 39(1): 108-113.

CHEN Qian, LI Chao, YANG Chengcheng, MU Bin, GONG Huan, HUANG Chao. Construction of Eukaryotic Expression Vector of Human Juxtaposed with Another Zincfinger Gene 1 and Its Influence on Embryonal Carcinoma Cell Apoptosis[J]. Journal of Sichuan Agricultural University, 2021, 39(1): 108-113.

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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

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