As the rate-limit enzyme of the pentose phosphate pathway, glucose-6-phosphate dehydrogenase (G6PD) plays important roles in tumour progression, but the exact mechanism through which G6PD controls cancer metastasis remains unclear.
G6PD expression in resected oral squamous cell carcinoma (OSCC) samples was analysed by immunohistochemistry. The effects and mechanism of G6PD suppression on OSCC cell lines were measured by transwell assay, wound healing assay, western and lectin blot, mass spectrometer analysis, ChIP-PCR, and luciferase reporter assay. BALB/c-nude mice were used to establish orthotopic xenograft model.
G6PD expression in the tumours of 105 OSCC patients was associated with lymphatic metastasis and prognosis. In vitro cellular study suggested that G6PD suppression impaired cell migration, invasion, and epithelial-mesenchymal transition. Furtherly, G6PD knockdown activated the JNK pathway, which then blocked the AKT/GSK-3β/Snail axis to induce E-Cadherin expression and transcriptionally regulated MGAT3 expression to promote bisecting GlcNAc-branched N-glycosylation of E-Cadherin. An orthotopic xenograft model further confirmed that dehydroepiandrosterone reduced lymphatic metastatic rate of OSCC, which was partially reversed by JNK inhibition.
Suppression of G6PD promoted the expression and bisecting GlcNAc-branched N-glycosylation of E-Cadherin via activating the JNK pathway, which thus acted on OSCC metastasis.
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Fong, M. Y., Zhou, W., Liu, L., Alontaga, A. Y., Chandra, M., Ashby, J. et al. Breast-cancer-secreted miR-122 reprograms glucose metabolism in premetastatic niche to promote metastasis. Nat Cell Biol 17, 183–194 (2015).
McDonald, O. G., Li, X., Saunders, T., Tryggvadottir, R., Mentch, S. J., Warmoes, M. O. et al. Epigenomic reprogramming during pancreatic cancer progression links anabolic glucose metabolism to distant metastasis. Nat Genet 49, 367–376 (2017).
Huang, R. & Zong, X. Aberrant cancer metabolism in epithelial-mesenchymal transition and cancer metastasis: mechanisms in cancer progression. Crit. Rev. Oncol. Hematol. 115, 13–22 (2017).
Patra, K. C. & Hay, N. The pentose phosphate pathway and cancer. Trends Biochem. Sci. 39, 347–354 (2014).
Riganti, C., Gazzano, E., Polimeni, M., Aldieri, E. & Ghigo, D. The pentose phosphate pathway: an antioxidant defense and a crossroad in tumor cell fate. Free Radic. Biol. Med. 53, 421–436 (2012).
Stanton, R. C. Glucose-6-phosphate dehydrogenase, NADPH, and cell survival. IUBMB Life 64, 362–369 (2012).
Rao, X., Duan, X., Mao, W., Li, X., Li, Z., Li, Q. et al. O-GlcNAcylation of G6PD promotes the pentose phosphate pathway and tumor growth. Nat. Commun. 6, 8468 (2015).
Pham, C. G., Bubici, C., Zazzeroni, F., Papa, S., Jones, J., Alvarez, K. et al. Ferritin heavy chain upregulation by NF-kappaB inhibits TNFalpha-induced apoptosis by suppressing reactive oxygen species. Cell 119, 529–542 (2004).
Almasi, S., Long, C. Y., Sterea, A., Clements, D. R., Gujar, S. & El, H. Y. TRPM2 silencing causes G2/M arrest and apoptosis in lung cancer cells via increasing intracellular ROS and RNS levels and activating the JNK pathway. Cell Physiol. Biochem. 52, 742–757 (2019).
Kalluri, R. & Neilson, E. G. Epithelial-mesenchymal transition and its implications for fibrosis. J. Clin. Invest. 112, 1776–1784 (2003).
Kalluri, R. & Weinberg, R. A. The basics of epithelial-mesenchymal transition. J. Clin. Invest. 119, 1420–1428 (2009).
Diepenbruck, M. & Christofori, G. Epithelial-mesenchymal transition (EMT) and metastasis: yes, no, maybe? Curr. Opin. Cell Biol. 43, 7–13 (2016).
Onder, T. T., Gupta, P. B., Mani, S. A., Yang, J., Lander, E. S. & Weinberg, R. A. Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 68, 3645–3654 (2008).
Lamouille, S., Xu, J. & Derynck, R. Molecular mechanisms of epithelial-mesenchymal transition. Nat. Rev. Mol. Cell Biol. 15, 178–196 (2014).
Oliveira-Ferrer, L., Legler, K. & Milde-Langosch, K. Role of protein glycosylation in cancer metastasis. Semin. Cancer Biol. 44, 141–152 (2017).
Pinho, S. S., Seruca, R., Gartner, F., Yamaguchi, Y., Gu, J., Taniguchi, N. et al. Modulation of E-cadherin function and dysfunction by N-glycosylation. Cell Mol. Life Sci. 68, 1011–1020 (2011).
Bray, F., Ferlay, J., Soerjomataram, I., Siegel, R. L., Torre, L. A. & Jemal, A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 68, 394–424 (2018).
Sturgis E. M., Ferlay J., Hashibe M., Winn D. M. Oral cavity, Oropharynx, Lip, and Salivary Glands. in Cancer epidemiology and prevention (eds Linet M. S., Cerhan J. R., Thun M. J., Haiman C. A., D, S), 543–578. (Oxford University Press, New York, 2018)
Kreppel, M., Nazarli, P., Grandoch, A., Safi, A. F., Zirk, M., Nickenig, H. J. et al. Clinical and histopathological staging in oral squamous cell carcinoma - Comparison of the prognostic significance. Oral Oncol. 60, 68–73 (2016).
Koyfman, S. A., Ismaila, N., Crook, D., D’Cruz, A., Rodriguez, C. P., Sher, D. J. et al. Management of the neck in squamous cell carcinoma of the oral cavity and oropharynx: ASCO clinical practice guideline. J. Clin. Oncol. 37, 1753–1774 (2019).
Pu, Y. F., Wang, L., Wu, H. H., Bian, H., Hong, Y. Y., Wang, Y. X. et al. Generation of homologous cell pairs using the oral lymphatic system. Int. J. Clin. Exp. Pathol. 7, 1563–1571 (2014).
Bais, M. V., Kukuruzinska, M. & Trackman, P. C. Orthotopic non-metastatic and metastatic oral cancer mouse models. Oral Oncol. 51, 476–482 (2015).
Xiang, G., Li, X., Cao, L., Zhu, C., Dai, Z., Pan, S. et al. Frequent overexpression of PDK1 in primary nasopharyngeal carcinoma is associated with poor prognosis. Pathol. Res. Pract. 212, 1102–1107 (2016).
Shi, Y., Nikulenkov, F., Zawacka-Pankau, J., Li, H., Gabdoulline, R., Xu, J. et al. ROS-dependent activation of JNK converts p53 into an efficient inhibitor of oncogenes leading to robust apoptosis. Cell Death Differ. 21, 612–623 (2014).
Zhou, B. P., Deng, J., Xia, W., Xu, J., Li, Y. M., Gunduz, M. et al. Dual regulation of Snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition. Nat. Cell Biol. 6, 931–940 (2004).
Bachelder, R. E., Yoon, S. O., Franci, C., de Herreros, A. G. & Mercurio, A. M. Glycogen synthase kinase-3 is an endogenous inhibitor of Snail transcription: implications for the epithelial-mesenchymal transition. J. Cell Biol. 168, 29–33 (2005).
Boroughs, L. K. & DeBerardinis, R. J. Metabolic pathways promoting cancer cell survival and growth. Nat. Cell Biol. 17, 351–359 (2015).
Warburg, O. On respiratory impairment in cancer cells. Science 124, 269–270 (1956).
Vander, H. M., Cantley, L. C. & Thompson, C. B. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324, 1029–1033 (2009).
Payen, V. L., Porporato, P. E., Baselet, B. & Sonveaux, P. Metabolic changes associated with tumor metastasis, part 1: tumor pH, glycolysis and the pentose phosphate pathway. Cell Mol. Life Sci. 73, 1333–1348 (2016).
Ma, X., Wang, L., Huang, D., Li, Y., Yang, D., Li, T. et al. Polo-like kinase 1 coordinates biosynthesis during cell cycle progression by directly activating pentose phosphate pathway. Nat. Commun. 8, 1506 (2017).
Wu, S., Wang, H., Li, Y., Xie, Y., Huang, C., Zhao, H. et al. Transcription factor YY1 promotes cell proliferation by directly activating the pentose phosphate pathway. Cancer Res. 78, 4549–4562 (2018).
Pes, G. M., Errigo, A., Soro, S., Longo, N. P. & Dore, M. P. Glucose-6-phosphate dehydrogenase deficiency reduces susceptibility to cancer of endodermal origin. Acta Oncol. 58, 1205–1211 (2019).
Srinivas, U. S., Tan, B., Vellayappan, B. A., Jeyasekharan, A. D. ROS and the DNA damage response in cancer. Redox Biol. 25, 101084 (2018).
Fukawa, T., Kajiya, H., Ozeki, S., Ikebe, T. & Okabe, K. Reactive oxygen species stimulates epithelial mesenchymal transition in normal human epidermal keratinocytes via TGF-beta secretion. Exp. Cell Res. 318, 1926–1932 (2012).
Li, H. Y., Zhang, J., Sun, L. L., Li, B. H., Gao, H. L., Xie, T. et al. Celastrol induces apoptosis and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells: an in vitro and in vivo study. Cell Death Dis. 6, e1604 (2015).
Pinho, S. S., Osorio, H., Nita-Lazar, M., Gomes, J., Lopes, C., Gartner, F. et al. Role of E-cadherin N-glycosylation profile in a mammary tumor model. Biochem. Biophys. Res. Commun. 379, 1091–1096 (2009).
Zhao, Y., Nakagawa, T., Itoh, S., Inamori, K., Isaji, T., Kariya, Y. et al. N-acetylglucosaminyltransferase III antagonizes the effect of N-acetylglucosaminyltransferase V on alpha3beta1 integrin-mediated cell migration. J. Biol. Chem. 281, 32122–32130 (2006).
Yoshimura, M., Nishikawa, A., Ihara, Y., Taniguchi, S. & Taniguchi, N. Suppression of lung metastasis of B16 mouse melanoma by N-acetylglucosaminyltransferase III gene transfection. Proc. Natl Acad. Sci. USA 92, 8754–8758 (1995).
Pinho, S. S., Reis, C. A., Paredes, J., Magalhaes, A. M., Ferreira, A. C., Figueiredo, J. et al. The role of N-acetylglucosaminyltransferase III and V in the post-transcriptional modifications of E-cadherin. Hum. Mol. Genet. 18, 2599–2608 (2009).
Huber, A. H. & Weis, W. I. The structure of the beta-catenin/E-cadherin complex and the molecular basis of diverse ligand recognition by beta-catenin. Cell 105, 391–402 (2001).
Helenius, A. & Aebi, M. Intracellular functions of N-linked glycans. Science 291, 2364–2369 (2001).
Sengupta, P. K., Bouchie, M. P. & Kukuruzinska, M. A. N-glycosylation gene DPAGT1 is a target of the Wnt/beta-catenin signaling pathway. J. Biol. Chem. 285, 31164–31173 (2010).
Nita-Lazar, M., Noonan, V., Rebustini, I., Walker, J., Menko, A. S. & Kukuruzinska, M. A. Overexpression of DPAGT1 leads to aberrant N-glycosylation of E-cadherin and cellular discohesion in oral cancer. Cancer Res. 69, 5673–5680 (2009).
Jamal, B., Sengupta, P. K., Gao, Z. N., Nita-Lazar, M., Amin, B., Jalisi, S. et al. Aberrant amplification of the crosstalk between canonical Wnt signaling and N-glycosylation gene DPAGT1 promotes oral cancer. Oral Oncol. 48, 523–529 (2012).
We thank Yixiang Wang of the Central Laboratory and Jianyun Zhang of the Department of pathology of Peking University School and Hospital of Stomatology for the assistances in various aspects of this work.
Ethics approval and consent to participate
For the study on the expression of G6PD in the samples of OSCC patients, it was approved by the Ethics Committee of Peking University School and Hospital of Stomatology (NO. PKUSSIRB-201944044) in accordance with the Declaration of Helsinki. Informed consent was obtained from all patients, and telephone follow-up was performed to analyse the survival trends among these patients. For the animal study, all animal studies (including the mice euthanasia procedure) were done in compliance with the regulations and guidelines of Peking University institutional animal care and conducted according to the AAALAC and the IACUC guidelines (NO. LA2019011).
The data that support the findings of this study are available from the corresponding author upon reasonable request.
The authors declare no competing interests.
This study was supported by National Natural Science Foundation of China (81972540, 81672664, and 81802699), and the fund of Peking University School and Hospital of Stomatology (PKUSS20170206).
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Wang, Y., Li, Q., Niu, L. et al. Suppression of G6PD induces the expression and bisecting GlcNAc-branched N-glycosylation of E-Cadherin to block epithelial-mesenchymal transition and lymphatic metastasis. Br J Cancer (2020). https://doi.org/10.1038/s41416-020-1007-3