中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2011年
6期
532-536
,共5页
牛志国%余志豪%陈丽媛%黄青松%高攀%何钰辉%王辉
牛誌國%餘誌豪%陳麗媛%黃青鬆%高攀%何鈺輝%王輝
우지국%여지호%진려원%황청송%고반%하옥휘%왕휘
T淋巴细胞%HTLV-1%EGR-1%Tax%NF-κB
T淋巴細胞%HTLV-1%EGR-1%Tax%NF-κB
T림파세포%HTLV-1%EGR-1%Tax%NF-κB
T lymphocytes%HTLV-1%EGR-1%Tax%NF-κB
目的 观察人T细胞白血病病毒1型(HTLV-1)的Tax蛋白阳性细胞中早期生长反应基因-1(EGR-1)与NF-κB的关系.方法 RT-PCR扩增MT2细胞中EGR-1的cDNA全长,连接于真核表达载体pcDNA3.0;用脂质体介导的方法将pcDNA3.0-EGR-1转染TaxP/TaxN细胞,48 h后PCR检测EGR-1和p65 mRNA的表达,Western blot检测EGR-1及p65蛋白的表达;将pcDNA3.0-EGR-1和NF-κB报告基因共转染TaxP及TaxN细胞48 h后检测荧光素酶活性.结果 成功构建了重组表达质粒pcDNA3.0-EGR-1,Tax可以促进EGR-1 mRNA和蛋白的表达;EGR-1可以促进TaxP细胞p65 mRNA和蛋白的表达,上调NF-κB活性.结论 EGR-1可能通过促进NF-κB活化参与成人T淋巴细胞白血病(adult T-cell leukemia,ATL)的发病.
目的 觀察人T細胞白血病病毒1型(HTLV-1)的Tax蛋白暘性細胞中早期生長反應基因-1(EGR-1)與NF-κB的關繫.方法 RT-PCR擴增MT2細胞中EGR-1的cDNA全長,連接于真覈錶達載體pcDNA3.0;用脂質體介導的方法將pcDNA3.0-EGR-1轉染TaxP/TaxN細胞,48 h後PCR檢測EGR-1和p65 mRNA的錶達,Western blot檢測EGR-1及p65蛋白的錶達;將pcDNA3.0-EGR-1和NF-κB報告基因共轉染TaxP及TaxN細胞48 h後檢測熒光素酶活性.結果 成功構建瞭重組錶達質粒pcDNA3.0-EGR-1,Tax可以促進EGR-1 mRNA和蛋白的錶達;EGR-1可以促進TaxP細胞p65 mRNA和蛋白的錶達,上調NF-κB活性.結論 EGR-1可能通過促進NF-κB活化參與成人T淋巴細胞白血病(adult T-cell leukemia,ATL)的髮病.
목적 관찰인T세포백혈병병독1형(HTLV-1)적Tax단백양성세포중조기생장반응기인-1(EGR-1)여NF-κB적관계.방법 RT-PCR확증MT2세포중EGR-1적cDNA전장,련접우진핵표체재체pcDNA3.0;용지질체개도적방법장pcDNA3.0-EGR-1전염TaxP/TaxN세포,48 h후PCR검측EGR-1화p65 mRNA적표체,Western blot검측EGR-1급p65단백적표체;장pcDNA3.0-EGR-1화NF-κB보고기인공전염TaxP급TaxN세포48 h후검측형광소매활성.결과 성공구건료중조표체질립pcDNA3.0-EGR-1,Tax가이촉진EGR-1 mRNA화단백적표체;EGR-1가이촉진TaxP세포p65 mRNA화단백적표체,상조NF-κB활성.결론 EGR-1가능통과촉진NF-κB활화삼여성인T림파세포백혈병(adult T-cell leukemia,ATL)적발병.
Objective To research the relation of early growth response gene-1(EGR-1) and NF-κB in human T-cell leukemia virus type 1(HTLV-1) Tax protein positive cells. Methods RT-PCR was used to amplify the aimed segments EGR-1 cDNA which was then inserted into an eukaryotic expression plasmid pcDNA3.0 to construct pcDNA3.0-EGR-1. The constructed plasmid was transfected into TaxN and TaxP cells by Tfx-50-mediated transfer method, the expression levels of EGR-1, p65 and Tax mRNA in transfected cells were assay by RT-PCR after 48 h post-transfection, the proteins of EGR-1 and p65 were detected by Western blot after 48 h post-transfection too. The constructed plasmid and pNF-κB-luc reporter gene plasmid was co-transfected into TaxN and TaxP cells by Tfx-50-mediated transfer method, and the activity of luciferase was assay after 48 h post-transfection. Results The results showed that the eukaryotic expression plasmid pcDNA3.0-EGR-1 was successfully constructed. The mRNA and protein expression of EGR-1 could be promoted significantly by Tax. EGR-1 can promote the mRNA and protein expressions of p65 in TaxP cells, the activity of NF-κB was up-regulated by EGR-1 too. Conclusion EGR-1 maybe involve in adult T-cell leukemia(ATL) by increasing the activation of NF-κB.
