Abstract P2-02-12: Gain-of-function of mutant P53 elevates glucose uptake via membrane translocation of GLUT1 in breast cancer
Authors: Ahn SG , Bae SJ , Yoon CI , Cha C , Park SE , Jeong J 
Published: May 2019

Background

High rate of glycolysis exhibited by cancer cells plays a crucial role in carcinogenesis and tumor progression. Previous study showed that gain-of-function (GOF) by mutant P53 drives the Warburg effect via GLUT1 membrane translocation in malignancies. Also, in the patients with a preoperative 18F-fluorodeoxy glucose positron emission tomography (FDG-PET), which enables to estimate glucose uptake by tumor as standardize uptake value (SUV), we identified high SUV as a poor prognostic marker. In this study, we wondered whether P53 mutation promotes GLUT1 membrane translocation and increases glucose uptake in MCF10A cells and patients with breast cancer.

Methods

Transcriptomic profiling was carried out using gene expression data from 66 breast cancer patients who underwent preoperative FDG-PET to identify a molecular signature associated with SUV. Gene network analyses revealed dysregulation of P53 pathway. We generated both MCF10A cell lines with P53-knock down and P53-mutant which include sequence variants (R175H, R273H, and R248W). Glucose uptake assay and confocal imaging studies with 2NDBG were performed. Confocal imaging system was used to detect GLUT1 membrane expression. In case-matched patients according to P53 mutation (n=114), we compared SUV. Mutational analysis of exons 5-9 of the P53 gene was carried out using Sanger sequencing.

Results

Glucose uptake assay showed that the level of glucose uptake was increased in P53 mutant MCF10A cell lines with a 1.5-1.8 fold change compare with wild-type MCF10A. The increase of glucose uptake by mutant-P53 was reproducible with the experiments with 2NBGD. By contrast, stable knock down of P53 did not induce an elevation of glucose uptake in MCF10A cells. The confocal imaging studies captured translocation to membrane of GLUT1 in mutant P53-MCF10A. However, in wild-type MCF10A, GLUT1 was diffusely expressed in cytoplasm. . In tumors from patients, membraneous expression of GLUT1 was significantly higher in tumors with p53 mutation than in tumors with intact p53 (P=0.022). Further, the mean SUV of p53-mutational group was significantly higher than that of wild-type p53-group (7.49 vs. 5.44, P=0.013). In multivariate analysis for recurrence-free survival, p53 mutational status carried prognostic significance (hazard ratio 3.73, 95% CI 1.15-12.07) independent of tumor size, nodal status, and estrogen receptor status.

Conclusion

We showed that GOF by P53 mutation promotes GLUT1 translocation to membrane, which consequently induces glucose influx in MCF10A cells, elucidating that P53 mutation contributes to the increase of SUV in patients with breast cancer.

Citation Format: Ahn SG, Bae SJ, Yoon CI, Cha C, Park SE, Jeong J. Gain-of-function of mutant P53 elevates glucose uptake via membrane translocation of GLUT1 in breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-02-12.