title: [Effects of eEF1A1 re-expression on proliferation and apoptosis of Jurkat cells with knocked down eEF1A1 gene and its mechanisms].
abstract: This study was aimed to explore the effects of expressing eukaryotic elongation factor 1A1 (eEF1A1) on proliferation and apoptosis in human acute T lymphocytic leukemia (T-ALL) cell line Jurkat with knocked down eEF1A1 gene and its mechanisms. eEF1A1-expressing lentivirus (LV) was constructed and used to transfect the Jurkat cells with knocked down eEF1A1 gene. Then, the expressions of eEF1A1 mRNA and protein were detected by real time PCR(RT-PCR) and Western blot respectively.Cell proliferation, apoptosis and cycle were detected by MTT method, Annexin V-APC labeling and DNA ploidy analysis respectively. The related protein expressions of phosphatidylinositol-3-kinase (PI3K)/serine/threonine kinase (Akt) signaling pathway were detected by Western blot. The results indicated that eEF1A1 mRNA and protein expressions of Jurkat cells with knocked down eEF1A1 gene were re-established by constructing eEF1A1-expression LV. Compared with negative control group (transfected with negative control LV and eEF1A1-shRNA LV), cell proliferation in eEF1A1 expression group was significantly enhanced, cell apoptosis was remarkably inhibited, percentage of cells in G0/G1 phase was significantly reduced alone with increased percentage of cells in S and G2/M phase, and the expression levels of p-Akt (Ser 473), nuclear factor kappa B (NF-κB), p-NF-κB (Ser 468), mammalian target of rapamycin (mTOR) and p-mTOR (Ser 2448) protein significantly increased. It is concluded that eEF1A1 may have a carcinogenic effect in T-ALL cells. eEF1A1 expression has noticeable effects on the proliferation enhancement and apoptosis inhibition of Jurkat cells, which may be mediated by the up-regulation of PI3K/Akt/NF-κB and PI3K/Akt/ mTOR signaling pathway.
authors.affiliation: Fujian Medical University, Fuzhou, Fujian Province, China. Hyi8070@126.com
authors.forename: Yi
authors.initials: Y
authors.lastname: Huang
authors.forename: Jian-Da
authors.initials: JD
authors.lastname: Hu
authors.forename: Yan-An
authors.initials: YA
authors.lastname: Wu
authors.forename: Jing
authors.initials: J
authors.lastname: Zheng
authors.forename: Yuan-Lin
authors.initials: YL
authors.lastname: Qi
authors.forename: Ying-Yu
authors.initials: YY
authors.lastname: Chen
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Huang
mesh_terms: D017209:Apoptosis; D049109:Cell Proliferation; D015870:Gene Expression; D006801:Humans; D019169:Jurkat Cells; D020648:Peptide Elongation Factor 1 / Q000235:genetics* / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction
chemical_list: C094814:EEF1A1 protein, human; D020648:Peptide Elongation Factor 1; D034741:RNA, Small Interfering; D051057:Proto-Oncogene Proteins c-akt