The effect of the targeted silencing of anthrax toxin receptor 1 gene mediated by lentivirus on the expression of γ-globin gene
XU Qin, LI Gui-fang, JIN Ting-ting, LI Di, XIONG Yong-hong, AN Bang-quan, HUANG Shengwen
(1.Department of Clinical Laboratory, Guizhou Provincial People's Hospital, Guizhou,Guiyang 550002, China; 2. Department of Oncology, Tongren People's Hospital, Gui zhou, Tongren 554300, China; 3. Medical School of Guizhou University, Gui zhou, Guiyang 550025, China
Abstract: 【Abstract】 Objective To explore the effect of silent anthrax toxin receptor 1 (ANTXR1) gene on the expression of γ-globin gene. Methods A shRNA recombinant plasmid containing the ANTXR1 gene-specific sequence anda negative control recombinant plasmid were constructed. After restriction enzyme digestion and sequencing identification, the interfering vector and lentiviral plasmids were co-transfected into 293T cells and the virus supernatant was harvested. The virus recombinant plasmid vector was transfected into K562 cells and the stable cell line was screened by puromycin. The expression levels of ANTXR1 and γ globin gene of RNA in each group were detected by real time-PCR (RT-PCR) and the expression levels of γ globin in each group were detected by Western blotting. Statistical analysis was performed by t-test. Results Stably transfected K562 cell line with effectively interfered by ANTXR1 gene was obtained. RT-PCR results showed that in the ANTXR1- shRNA group, the relative expression levels of ANTXR1 gene and γ-globin gene were 0.19±0.02 and 1.46±0.24 respectively at mRNA level. Western blotting results showed that the relative level of protein were 0.20±0.01 and 1.45±0.14 respectively . There were significant differences in ANTXR1-shRNA group vs the negative control group, blank plasmid group and the blank control group, the expression level of ANTXR1 gene with down-regulation and the γ-globin gene with up-regulation (P ﹤ 0.05). Conclusion Targeted silencing of the ANTXR1 gene promotes the expression of γ-globin gene at mRNA and protein level.
徐琴 李贵芳 金婷婷 李頔 熊永红 安邦权 黄盛文. 慢病毒介导炭疽毒素受体1 基因靶向沉默对γ 珠蛋白基因表达的影响[J]. 发育医学电子杂志, 2018, 6(4): 213-219.
XU Qin, LI Gui-fang, JIN Ting-ting, LI Di, XIONG Yong-hong, AN Bang-quan, HUANG Shengwen. The effect of the targeted silencing of anthrax toxin receptor 1 gene mediated by lentivirus on the expression of γ-globin gene. Journal of Developmental Medicine(Electronic Version), 2018, 6(4): 213-219.
2018, Vol. 6 
