The online version of this article (https://doi.org/10.1186/s13045-018-0674-5) contains supplementary material, which is available to authorized users.
Wei Jiang, Tiancong He and Shuai Liu contributed equally to this work.
The study aims to present the effect of PIK3CA E542K and E545K mutations on glucose metabolism and proliferation and identify their underlying mechanisms in cervical cancer.
The maximum standard uptake value (SUVmax) of tumors was detected by18F-FDG PET/CT scan. In vitro, glycolysis analysis, extracellular acidification rate analysis, and ATP production were used to evaluate the impact of PIK3CA E542K and E545K mutations on glucose metabolism. The expression level of key glycolytic enzymes was evaluated by western blotting and immunohistochemical staining in cervical cancer cells and tumor tissues, respectively. Immunofluorescence analysis was used to observe the nuclear translocation of β-catenin. The target gene of β-catenin was analyzed by using luciferase reporter system. The glucose metabolic ability of the xenograft models was assessed by SUVmax from microPET/CT scanning.
Cervical cancer patients with mutant PIK3CA (E542K and E545K) exhibited a higher SUVmax value than those with wild-type PIK3CA (P = 0.037), which was confirmed in xenograft models. In vitro, enhanced glucose metabolism and proliferation was observed in SiHa and MS751 cells with mutant PIK3CA. The mRNA and protein expression of key glycolytic enzymes was increased. AKT/GSK3β/β-catenin signaling was highly activated in SiHa and MS751 cells with mutant PIK3CA. Knocking down β-catenin expression decreased glucose uptake and lactate production. In addition, the nuclear accumulation of β-catenin was found in SiHa cells and tumors with mutant PIK3CA. Furthermore, β-catenin downregulated the expression of SIRT3 via suppressing the activity of the SIRT3 promotor, and the reduced glucose uptake and lactate production due to the downregulation of β-catenin can be reversed by the transfection of SIRT3 siRNA in SiHa cells with mutant PIK3CA. The negative correlation between β-catenin and SIRT3 was further confirmed in cervical cancer tissues.
These findings provide evidence that the PI3K E542K and E545K/β-catenin/SIRT3 signaling axis regulates glucose metabolism and proliferation in cervical cancers with PIK3CA mutations, suggesting therapeutic targets in the treatment of cervical cancers.
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Additional file 1: Table S1. The primer sequences are used for qRT-PCR detection. (DOCX 15 kb)13045_2018_674_MOESM1_ESM.docx
Additional file 2: Figure S1. Relative mRNA expression of other glycolytic enzymes in SiHa and MS751 cells with wild-type and mutant PIK3CA. A: Relative mRNA expression of other key glycolytic enzymes in SiHa cells with wild-type and mutant PIK3CA. B: Relative mRNA expression of other key glycolytic enzymes in MS751 cells with wild-type and mutant PIK3CA. (TIF 1495 kb)13045_2018_674_MOESM2_ESM.tif
Additional file 3: Figure S2. The expression of other key glycolytic enzymes after knocking down the expression of β-catenin. (TIF 1169 kb)13045_2018_674_MOESM3_ESM.tif
Additional file 4: Figure S3. The correlation analysis between β-catenin and SIRT3 by western blotting in SiHa and MS751 cells with wild-type and mutant PIK3CA. (TIF 682 kb)13045_2018_674_MOESM4_ESM.tif
Additional file 5: Figure S4. The correlation analysis between β-catenin and SIRT3 in cervical cancer tissues with wild-type and mutant PIK3CA. (TIF 457 kb)13045_2018_674_MOESM5_ESM.tif
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- The PIK3CA E542K and E545K mutations promote glycolysis and proliferation via induction of the β-catenin/SIRT3 signaling pathway in cervical cancer
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