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SIRT6 promotes the Warburg effect of papillary thyroid cancer cell BCPAP through reactive oxygen species - PubMed

  • ️Tue Jan 01 2019

SIRT6 promotes the Warburg effect of papillary thyroid cancer cell BCPAP through reactive oxygen species

Weiping Yu et al. Onco Targets Ther. 2019.

Abstract

Purpose: Our previous study demonstrated that SIRT6 is upregulated in papillary thyroid cancer (PTC) and enhances tumor aggressiveness. In this study, we further researched its influence in the Warburg effect. Methods: SIRT6-upregulated and downregulated BCPAP cells and negative control BCPAP-NC groups were generated with lentiviral vectors. In these two cell lines, reactive oxygen species (ROS) were detected by dichlorodihydrofluorescein diacetate. Expression of the key Warburg effect genes including GLUT1, HK2, GAPDH, PGK1, ENO1, PKM2 and LDHA was measured by quantitative real-time PCR and western blotting. Glucose uptake, lactate production and the ATP content of cells were detected with assay kits. The ROS scavenger N-acetylcysteine was used for treatment of BCPAP-SIRT6, and the same measurements as described above were detected again. Results: Compared with the BCPAP-NC group, expression of the key Warburg effect genes including Glut1, HK2 and GAPDH and their protein products was upregulated in the BCPAP-SIRT6 group, whereas BCPAP-shSIRT6 showed significant downregulation. Meanwhile, ROS, glucose uptake, lactate production and ATP content of the BCPAP-SIRT6 group were also significantly increased, and BCPAP-shSIRT6 showed significant downregulation. Furthermore, upregulation of key Warburg effect genes and glucose uptake, lactate production and ATP content were all rescued after treatment with ROS scavenger. Conclusion: SIRT6 promoted the Warburg effect of PTC cells via upregulation of ROS. Inhibition of ROS in SIRT6-upregulated cells could rescue activation of the Warburg effect.

Keywords: Warburg effect; papillary thyroid cancer; reactive oxygen species; sirtuin 6.

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Conflict of interest statement

The author reports no conflicts of interest in this work.

Figures

Figure 1
Figure 1

Co-expression between SIRT6 and key Warburg effect genes.

Figure 2
Figure 2

(A) Protein expression of SIRT6 in BCPAP cells was detected by western blotting. (B) mRNA expression of SIRT6 in BCPAP cells was detected by quantitative real-time PCR, and the results were analyzed. (C) Glut1 protein expression in BCPAP cells was detected by western blotting. (D) Key Warburg effect gene expression in BCPAP cells was analyzed. Notes: All group values are given as mean±SD. Compared with BCPAP-NC: *P<0.05, **P<0.01 and ***P<0.001.

Figure 3
Figure 3

(A) Glucose uptake of BCPAP cells was detected and analyzed from cell culture medium. (B) Lactate production of BCPAP cells was detected and analyzed from cell culture medium. (C) ATP content of BCPAP cells was detected and analyzed by ATPLite luminescent assay. Notes: All group values are given as mean±SD. Compared with BCPAP-NC: *P<0.05, **P<0.01 and ***P<0.001.

Figure 4
Figure 4

(A) c-MYC and p53 protein expression was detected by western blotting. (B) ROS production measured by dichlorodihydrofluorescein diacetate, and the result was analyzed. Notes: The value is given as mean±SD . ***P<0.001. Abbreviation: ROS, reactive oxygen species.

Figure 5
Figure 5

(A) NAC could obviously inhibit ROS in BCPAP-SIRT6. (B) protein expression of Glut1 was rescued in BCPAP-SIRT6. (C) mRNA expression of PKM, Glut1, HK2, LDHA, Eno1, PGK1 and GAPDH was rescued in the BCPAP-SIRT6 group. Notes: All group values are given as mean±SD. *P<0.05, **P<0.01 and ***P<0.001. Abbreviations: NAC, N-acetylcysteine; ROS, reactive oxygen species.

Figure 6
Figure 6

(A) Glucose uptake was detected and analyzed from cell culture medium. (B) Lactate production was detected and analyzed from cell culture medium. (C) ATP content was detected and analyzed by ATPLite luminescent assay. Notes: All group values are given as mean±SD. *P<0.05, **P<0.01 and ***P<0.001. Abbreviation: NAC, N-acetylcysteine.

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