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Warburg effect regulated by amphiregulin in the development of colorectal cancer.

Nam SO, Yotsumoto F, Miyata K, Fukagawa S, Yamada H, Kuroki M, Miyamoto S - Cancer Med (2015)

Bottom Line: Upregulated (>2.0-fold) and downregulated (<0.5-fold) genes in 3DC compared with 2DC were selected.The suppression of AREG expression reduced the uptake of glucose and production of lactate.Together these data suggest that AREG plays a pivotal role in the development of CRC through activation of the Warburg effect.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Faculty of Medicine, Fukuoka University, Fukuoka, Japan; Central Research Institute for Advanced Molecular Medicine, Fukuoka University, Fukuoka, Japan.

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The transcriptional regulatory element of amphiregulin (AREG). (A) Dual luciferase reporter assays in HCT116 cells transfected with AREG promoter constructs. Activation of the reporter gene was calculated by the ratio of firefly luciferase activity to Renilla luciferase activity. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −180/+210, −380/+210, −680/+210, or −840/+210 constructs. (B) Dual luciferase reporter assays using a second set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/−40, −380/−80, or −380/−130 constructs. (C) Dual luciferase reporter assays using a third set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/+210 or −180/+210 construct.
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fig03: The transcriptional regulatory element of amphiregulin (AREG). (A) Dual luciferase reporter assays in HCT116 cells transfected with AREG promoter constructs. Activation of the reporter gene was calculated by the ratio of firefly luciferase activity to Renilla luciferase activity. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −180/+210, −380/+210, −680/+210, or −840/+210 constructs. (B) Dual luciferase reporter assays using a second set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/−40, −380/−80, or −380/−130 constructs. (C) Dual luciferase reporter assays using a third set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/+210 or −180/+210 construct.

Mentions: To identify the transcriptional factors that directly regulate AREG expression, we examined the transcriptional region controlling AREG expression using a luciferase reporter assay. Approximately 1.05 kbp was cloned from the transcriptional start site (TSS) of the AREG gene, which is conserved among mammalian species, and luciferase reporter vectors containing various fragments of the cloned region were examined. Luciferase assays showed that reporter vectors containing the promoter fragments −180/+210 bp from the AREG TSS (pGL/AREG−180/+210) (Fig.3A) and −380/−130 bp from the AREG TSS (pGL/AREG−380/−130) (Fig.3B) exhibited a 20-fold increase in luciferase activity compared with that of pGL/AREG−40/+210 or pGL/AREG−380/−180, respectively.


Warburg effect regulated by amphiregulin in the development of colorectal cancer.

Nam SO, Yotsumoto F, Miyata K, Fukagawa S, Yamada H, Kuroki M, Miyamoto S - Cancer Med (2015)

The transcriptional regulatory element of amphiregulin (AREG). (A) Dual luciferase reporter assays in HCT116 cells transfected with AREG promoter constructs. Activation of the reporter gene was calculated by the ratio of firefly luciferase activity to Renilla luciferase activity. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −180/+210, −380/+210, −680/+210, or −840/+210 constructs. (B) Dual luciferase reporter assays using a second set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/−40, −380/−80, or −380/−130 constructs. (C) Dual luciferase reporter assays using a third set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/+210 or −180/+210 construct.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4402072&req=5

fig03: The transcriptional regulatory element of amphiregulin (AREG). (A) Dual luciferase reporter assays in HCT116 cells transfected with AREG promoter constructs. Activation of the reporter gene was calculated by the ratio of firefly luciferase activity to Renilla luciferase activity. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −180/+210, −380/+210, −680/+210, or −840/+210 constructs. (B) Dual luciferase reporter assays using a second set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/−40, −380/−80, or −380/−130 constructs. (C) Dual luciferase reporter assays using a third set of AREG promoter constructs. Data were measured in triplicate and represent mean ± SD. *P < 0.05 versus the −380/+210 or −180/+210 construct.
Mentions: To identify the transcriptional factors that directly regulate AREG expression, we examined the transcriptional region controlling AREG expression using a luciferase reporter assay. Approximately 1.05 kbp was cloned from the transcriptional start site (TSS) of the AREG gene, which is conserved among mammalian species, and luciferase reporter vectors containing various fragments of the cloned region were examined. Luciferase assays showed that reporter vectors containing the promoter fragments −180/+210 bp from the AREG TSS (pGL/AREG−180/+210) (Fig.3A) and −380/−130 bp from the AREG TSS (pGL/AREG−380/−130) (Fig.3B) exhibited a 20-fold increase in luciferase activity compared with that of pGL/AREG−40/+210 or pGL/AREG−380/−180, respectively.

Bottom Line: Upregulated (>2.0-fold) and downregulated (<0.5-fold) genes in 3DC compared with 2DC were selected.The suppression of AREG expression reduced the uptake of glucose and production of lactate.Together these data suggest that AREG plays a pivotal role in the development of CRC through activation of the Warburg effect.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Faculty of Medicine, Fukuoka University, Fukuoka, Japan; Central Research Institute for Advanced Molecular Medicine, Fukuoka University, Fukuoka, Japan.

Show MeSH
Related in: MedlinePlus