Effect of TSC2 gene expression downregulation by lentivirus induced RNA interference on U937 cell line and its mechanism
10.3760/cma.j.issn.0253-2727.2017.07.012
- VernacularTitle: 慢病毒介导的RNA干扰下调TSC2表达对白血病U937细胞系的影响及作用机制
- Author:
Zhifang XU
1
;
Haixia LIU
;
Yanhong TAN
;
Xiuhua CHEN
;
Fanggang REN
;
Yaofang ZHANG
;
Jianmei CHANG
;
Na ZHANG
;
Jinjun HU
;
Hongwei WANG
Author Information
1. Department of Hematology, the Second Hospital of Shanxi Medical University, Shanxi Key Laboratory of Molecular Diagnosis and Treatment of Blood Diseases, Taiyuan 030001, China
- Publication Type:Journal Article
- Keywords:
Gene, TSC2;
mTOR;
Leukemia;
Signaling pathway
- From:
Chinese Journal of Hematology
2017;38(7):612-617
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of biology and mTOR pathway activity of down-regulated TSC2 gene expression on U937 leukemia cells.
Methods:Gene expression was down-regulated by lentivirus induced RNA interference on TSC2 high expressed U937 cell line; the proliferation, apoptosis and differentiation were detected by CCK-8 assay, colony formation assay and flow cytometry; the gene expression level and protein kinase activity were detected by qRT-PCR and Western blot.
Results:Down-regulated expression of TSC2 gene promoted U937 cell proliferation and colony formation ability (P<0.05) . The proportion in G0/G1 phase of TSC2 down-regulated U937 cell was much lower than that of the control cells [ (52.53±3.75) % vs (75.10±4.33) %, t=6.829, P=0.002], the S phase [ (22.43±1.00) % vs (15.47±1.20) %, t=-5.581, P=0.019] and G2/M phase [ (25.03±4.34) % vs (14.33±0.91) %, t=-5.413, P=0.013] was remarkably higher than that of the control cells (P<0.05) . There were no statistically significant differences in cell apoptosis and differentiation (P>0.05) . Down-regulation of TSC2 led to the increased activity of mTOR, 4EBP1 and S6K1, but did not influence the activity of AKT. The expressions of proliferation related cyclinD1, c-myc and PTEN were also up-regulated after TSC2 silenced, but the expressions of P27KIP and BCL-XL were not changed.
Conclusion:Downregulation of TSC2 could promote the proliferation of U937 cells through up-regulation of mTOR activity.
