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Phenotypic Knockout of CXCR4 on Molt-4 with SDF-1α/54 Attached with KDEL

Phenotypic Knockout of CXCR4 on Molt-4 with SDF-1α/54 Attached with KDEL
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摘要 Objective ： To investigate the mechanism of phenotypic knockout of CXCR4 on T-cell leukemia cell line Molt-4 via SDF-1α/54/KDEL intrakine technology, which the mutant SDF-1α/54, human stromal cell-derived Faceor-1 （SDF-1α） was deleted its C- terminal α-helix and attached with a endoplasimc reticulum retention signal 4-peptide- KDEL encoding gene, so that retain the newly synthesized receptor CXCR4 within the Molt-4 cells endoplasmic reticulum. Methods： The recombinant vector pEGFP-C3/SDF- 1α/54/KDEL were transfected into Cos-7 cells by liposome, SDF-1α/54/KDEL fusion protein was confirmed with western blot. The recombinant plasmids were transfected transiently into Molt-4 by electroporation. Results：Western blot confirmed SDF-1α/54/KDEL expression in Cos-7. A dramatic downregulation of CXCR4 expression on Molt-4 was demonstrated by flow cytometric （FCM） analysis. Conelusion：SDF-1α/54/KDEL and SDF- 1αKDEL have no significant deviation for phenotypic knockout of CXCR4. These suggest that the phenotypic knockout effects of SDF-1α/54 against CXCR4 are not influenced by deleting of SDF-1α helix in the C-terminal. Objective:To investigate the mechanism of phenotypic knockout of CXCR4 on T-cell leukemia cell line Molt-4 via SDF-1α/54/KDEL intrakine technology, which the mutant SDF-1α/54, human stromal cell-derived Faceor-1(SDF-1α) was deleted its C-terminal α-helix and attached with a endoplasimc reticulum retention signal 4-peptide-KDEL encoding gene, so that retain the newly synthesized receptor CXCR4 within the Molt-4 cells endoplasmic reticulum. Methods:The recombinant vector pEGFP-C3/SDF-1α/54/KDEL were transfected into Cos-7 cells by liposome, SDF-1α/54/KDEL fusion protein was confirmed with western blot. The recombinant plasmids were transfected transiently into Molt-4 by electroporation. Results:Western blot confirmed SDF-1α/54/KDEL expression in Cos-7.A dramatic downregulation of CXCR4 expression on Molt-4 was demonstrated by flow cytometric (FCM) analysis. Conclusion:SDF-1α/54/KDEL and SDF-1αKDEL have no significant deviation for phenotypic knockout of CXCR4. These suggest that the phenotypic knockout effects of SDF-1α/54 against CXCR4 are not influenced by deleting of SDF-1α helix in the C-terminal.
作者 CHEN Hong-yuan TAN Yi GUO Zhi-gang MA Wei-feng CAI Shao-xi DU Jun HUANG Jun CAI Shao-hui
机构地区 Key Laboratory for Biomechanics ＆ Tissue Engineering of the State Ministry of Education Department of Microbiology-Immunology School of Guangdong Pharmacy University Department of Clinical Pharmacology School of Pharmaceutical Sciences School of Pharmaceutical Sciences Department of Microbiology-Immunology School of Sun Yat-Sen University
出处 《Chinese Journal of Biomedical Engineering(English Edition)》 2007年第3期111-116,共6页 中国生物医学工程学报（英文版）
基金 Grant sponsor:National Natural Science Foundation of China Grant number:30572209
关键词 stromal cell-derived Faceor-1 SDF-1α/54 CXCR4 phenotypic knockout 细胞学 细胞诱导因子 SDF-1α/54 CXCR4
分类号 Q2 [生物学—细胞生物学]
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Chinese Journal of Biomedical Engineering(English Edition)
2007年 第3期

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