Effect of siRNA‑mediated gene silencing of transketolase on A549 lung cancer cells

Lu,Huan; Zhu,Huili

doi:10.3892/ol.2017.6916

Effect of siRNA‑mediated gene silencing of transketolase on A549 lung cancer cells

Authors:
- Huan Lu
- Huili Zhu
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Affiliations: Department of Respiratory Medicine, Huadong Hospital, Fudan University, Shanghai 200040, P.R. China
Published online on: September 8, 2017 https://doi.org/10.3892/ol.2017.6916
Pages: 5906-5912
Copyright: © Lu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the effects of transketolase (TKT) on cell proliferation, cell migration and interaction with other metabolism‑associated genes in A549 lung cancer cells. A549 cells were transfected with three TKT‑specific small interfering (si)RNAs, screened for the optimal transfection concentration, and sequenced with flow cytometry and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Cell viability was evaluated using Cell Counting Kit‑8 (CCK‑8), cell cycle was assessed by flow cytometric analysis. Cell migration was determined by scratch‑wound and Transwell chamber assays. The changes in mRNA expression levels of glucose‑6‑phosphate dehydrogenase (G6PDH), transaldolase (TAL), sorbitol dehydrogenase (SORD), phosphoribosyl pyrophosphate synthetase 1 (PRPS1) and hexokinase 1 (HK1) were detected by RT‑qPCR. siRNA‑C at 50 nmol/l was selected for the subsequent experiments. Compared with the negative control, cell proliferation of the TKT‑siRNA‑C group was inhibited dramatically (CCK‑8 24 h, 0.2984±0.0371 vs. 0.0952±0.0063; P<0.0001), the cell cycle was arrested at the G1/G0 cell cycle phase (58±2.0% vs. 70±2.5%; P=0.002), and cell migration ability was decreased [wound size, 254.71±34.96 vs. 349.12±37.43 µm (P=0.0001); Transwell migration, 250±47.8/field vs. 150±49.0/field (P<0.0001)]. The mRNA expression levels of G6PDH, TAL, SORD, PRPS1 and HK1 were downregulated in the TKT‑siRNA‑C group compared with the negative control. The present study revealed that synthetic TKT‑siRNA can inhibit A549 cell viability and migration, which may be due to arrest of the cell cycle and downregulation of relevant metabolic enzymes.

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