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Lung Cancer: Lung Cancer II |

Effect of Gene Silencing of Transketolase Mediated by RNAi in A549 Lung Cancer Cells FREE TO VIEW

Huili Zhu, MD; Huan Lu, MD
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Huadong Hospital Affiliated to Fudan University, Shanghai, China


Copyright 2016, American College of Chest Physicians. All Rights Reserved.


Chest. 2016;149(4_S):A303. doi:10.1016/j.chest.2016.02.316
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SESSION TITLE: Lung Cancer II

SESSION TYPE: Original Investigation Poster

PRESENTED ON: Saturday, April 16, 2016 at 11:45 AM - 12:45 PM

PURPOSE: To study the effect of cell proliferation and apoptosis in A549 lung cancer cells and to find out the influence on other key enzymes of metabolism.

METHODS: With three chemosynthesis siRNA Duplex targeting transketolase (TKT), A549 lung cancer cells were cultured and transfected. One of the best siRNA with highest interference effect was selected by fluorogenic Quantitative PCR. Transfection cells of 48h and 72h were detected by Flow Cytometry for cell cycle and apoptosis and by fluorogenic Quantitative PCR to record the changes of other metabolic key enzymes mRNA.

RESULTS: The results showed that after transfection 48h the proportion of G1/G0 cells in Mock group and siRNA group were respectively and the proportion of G2/M+S cells in Mock group and siRNA group were respectively, no significant difference was found; After transfection 72h the proportion of G1/G0 cells in Mock group and siRNA group were respectively, the difference was statistically significant, and the proportion of G2/M+S cells in Mock group and siRNA group were respectively, the difference was statistically significant, P=0.002. Quantitative PCR results showed that after transfection 48h the relative expression of TKT mRNA decreased significantly while the relative expression of G6PDH, TAL and SORD mRNA didn’t change significantly; transfection 72h the relative expression of TKT mRNA decreased significantly and the relative expression of G6PDH, TAL, PRPS1, HK1 and SORD mRNA decreased significantly, the relative expression of HK2 mRNA didn’t change significantly.

CONCLUSIONS: Quantitative PCR results showed that the best interference effect was found in siRNA-A group (Negative control: siRNA groups ratio is 1: 0.35). TKT mRNA interference inhibit A549 cell proliferation, the proportion of G2/M+S cells decreases and cell cycle arrest in the G1/G0 stage.

CLINICAL IMPLICATIONS: TKT mRNA interference has a great influence on the normal metabolism and proliferation of A549 lung cancer cells. TKT may be a potential target for the treatment of tumor.

DISCLOSURE: The following authors have nothing to disclose: Huili Zhu, Huan Lu

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