Effect of TSC2 gene expression downregulation by lentivirus induced RNA interference on U937 cell line and its mechanism

• Published in: Zhōnghuá xuèyèxué zázhì Vol. 38; no. 7; pp. 612 - 617
• Main Authors: Xu, Z F, Liu, H X, Tan, Y H, Chen, X H, Ren, F G, Zhang, Y F, Chang, J M, Zhang, N, Hu, J J, Wang, H W
• Format: Journal Article
• Language: Chinese
• Published: China 14.07.2017
• Subjects: Apoptosis; Cell Proliferation; Down-Regulation; Humans; Lentivirus; RNA Interference; Tuberous Sclerosis Complex 2 Protein - genetics; U937 Cells; mTOR; Gene, TSC2; Signaling pathway; Leukemia
• ISSN: 0253-2727
• DOI: 10.3760/cma.j.issn.0253-2727.2017.07.012

To investigate the effect of biology and mTOR pathway activity of down-regulated TSC2 gene expression on U937 leukemia cells. 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. Down-regulated expression of TSC2 gene promoted U937 cell proliferation and colony formation ability ( <0.05) . The proportion in G(0)/G(1) phase of TSC2 down-regulated U937 cell was much lower than that of the control cells [ (52.53±3.75) % (75.10±4.33) %, =6.829, =0.002], the S phase [ (22.43±1.00) % (15.47±1.20) %, =-5.581, =0.019] and G(2)/M phase [ (25.03±4.34) % (14.33±0.91) %, =-5.413, =0.013] was remarkably higher than that of the control cells ( <0.05) . There were no statistically significant differences in cell apoptosis and differentiation ( >0.05)