Inhibition of PI3K/Akt signaling suppresses epithelial-to-mesenchymal transition in hepatocellular carcinoma through the Snail/GSK-3/beta-catenin pathway
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Background/Aims: Patients with advanced hepatocellular carcinoma (HCC) have a poor prognosis due to the lack of effective systemic therapies. Epithelial-to-mesenchymal transition (EMT) is a pivotal event in tumor progression, during which cancer cells acquire invasive properties. In this study, we investigated the effects of phosphatidylinositol 3-kinase (PI3K) inhibitors, including LY294002 and idelalisib, on the EMT features of HCC cells in vitro. Methods: Human HCC cell lines, including Huh-BAT and HepG2, were used in this study. Cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, and cell cycle distributions were evaluated using a flow cytometer by propidium iodide staining. Immunofluorescence staining, quantitative real-time polymerase chain reaction, and immunoblotting were performed to detect EMT-associated changes. Results: PI3K inhibitors suppressed the proliferation and invasion of HCC cells and deregulated the expression of EMT markers, as indicated by increased expression of E-cadherin, an epithelial marker, and decreased expression of N-cadherin, a mesenchymal marker, and Snail, a transcription factor implicated in EMT regulation. Furthermore, LY294002 and idelalisib inhibited the phosphorylation of GSK-3β and induced the nuclear translocation of GSK-3β, which corresponded to the downregulation of Snail and β-catenin expressions in Huh-BAT and HepG2 cells. Conclusions: The inhibition of PI3K/Akt signaling decreases Snail expression by enhancing the nuclear translocation of GSK-3β, which suppresses EMT in HCC cells, suggesting the potential clinical application of PI3K inhibitors for HCC treatment.
Abstract
Background/Aims: Patients with advanced hepatocellular carcinoma (HCC) have a poor prognosis due to the lack of effective systemic therapies. Epithelial-to-mesenchymal transition (EMT) is a pivotal event in tumor progression, during which cancer cells acquire invasive properties. In this study, we investigated the effects of phosphatidylinositol 3-kinase (PI3K) inhibitors, including LY294002 and idelalisib, on the EMT features of HCC cells in vitro. Methods: Human HCC cell lines, including Huh-BAT and HepG2, were used in this study. Cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, and cell cycle distributions were evaluated using a flow cytometer by propidium iodide staining. Immunofluorescence staining, quantitative real-time polymerase chain reaction, and immunoblotting were performed to detect EMT-associated changes. Results: PI3K inhibitors suppressed the proliferation and invasion of HCC cells and deregulated the expression of EMT markers, as indicated by increased expression of E-cadherin, an epithelial marker, and decreased expression of N-cadherin, a mesenchymal marker, and Snail, a transcription factor implicated in EMT regulation. Furthermore, LY294002 and idelalisib inhibited the phosphorylation of GSK-3β and induced the nuclear translocation of GSK-3β, which corresponded to the downregulation of Snail and β-catenin expressions in Huh-BAT and HepG2 cells. Conclusions: The inhibition of PI3K/Akt signaling decreases Snail expression by enhancing the nuclear translocation of GSK-3β, which suppresses EMT in HCC cells, suggesting the potential clinical application of PI3K inhibitors for HCC treatment.
참고문헌 (37)
- Siegel R, "Cancer statistics, 2013", CA Cancer J Clin, 63, pp.11~30 (2013)
- Thomas MB, "Hepatocellular carcinoma : the need for progress", J Clin Oncol, 23, pp.2892~2899 (2005)
- Ferenci P, "Hepatocellular carcinoma(HCC) : a global perspective", J Clin Gastroenterol, 44, pp.239~245 (2010)
- van Zijl F, "A human model of epithelial to mesenchymal transition to monitor drug efficacy in hepatocellular carcinoma progression", Mol Cancer Ther, 10, pp.850~860 (2011)
- Kalluri R, "The basics of epithelial-mesenchymal transition", J Clin Invest, 119, pp.1420~1428 (2009)
- Thiery JP, "Epithelial-mesenchymal transitions in development and disease", Cell, 139, pp.871~890 (2009)
- Wu Y, "Snail : more than EMT", Cell Adh Migr, 4, pp.199~203 (2010)
- Peinado H, "Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype?", Nat Rev Cancer, 7, pp.415~428 (2007)
- Nieto MA, "The snail superfamily of zinc-finger transcription factors", Nat Rev Mol Cell Biol, 3, pp.155~166 (2002)
- Prokop JW, "A method for in silico identification of SNAIL", SLUG DNA binding potentials to the E-box sequence using molecular dynamics and evolutionary conserved amino acids, 2013, pp.19, 3463~3469
- Batlle E, "The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells", Nat Cell Biol, 2, pp.84~89 (2000)
- Cano A, "The transcription factor snail controls epithelialmesenchymal transitions by repressing E-cadherin expression", Nat Cell Biol, 2, pp.76~83 (2000)
- Dong C, "G9a interacts with Snail and is critical for Snail-mediated E-cadherin repression in human breast cancer", J Clin Invest, 122, pp.1469~1486 (2012)
- McCubrey JA, "Roles of GSK-3 and microRNAs on epithelial mesenchymal transition and cancer stem cells", Oncotarget, 8, pp.14221~14250 (2017)
- Ahn SY, "Niclosamide is a potential therapeutic for familial adenomatosis polyposis by disrupting Axin-GSK3 interaction", Oncotarget, 8, pp.31842~31855 (2017)
- Burger JA, "Haematological cancer : idelalisib-targeting PI3Kδ in patients with B-cell malignancies", Nat Rev Clin Oncol, 11, pp.184~186 (2014)
- Henderson V, "Snail promotes cell migration through PI3K", AKT-dependent Rac1 activation as well as PI3K, Cell Adh Migr, pp.2015, 9[DOI]
- Zawel L, "P3Kα : a driver of tumor metastasis?", Oncotarget, 1, pp.315~316 (2010)
- Castillo JJ, "CAL-101 : a phosphatidylinositol-3-kinase p110-delta inhibitor for the treatment of lymphoid malignancies", Expert Opin Investig Drugs, 21, pp.15~22 (2012)
- Taylor JR, "Cooperative effects of Akt-1 and Raf-1 on the induction of cellular senescence in doxorubicin or tamoxifen treated breast cancer cells", Oncotarget, 2, pp.610~626 (2011)
- Kunter I, "Active form of AKT controls cell proliferation and response to apoptosis in hepatocellular carcinoma", Oncol Rep, 31, pp.573~580 (2014)
- Bakin AV, "Phosphatidylinositol 3-kinase function is required for transforming growth factor beta-mediated epithelial to mesenchymal transition and cell migration", J Biol Chem, 275, pp.36803~36810 (2000)
- Xu Q, "α-Mangostin suppresses the viability and epithelial-mesenchymal transition of pancreatic cancer cells by downregulating the PI3K", Akt pathwa, 2014, pp.2014, 546353~
- Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method", Methods, 25, pp.402~408 (2001)
- Zhou BP, "Dual regulation of Snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition", Nat Cell Biol, 6, pp.931~940 (2004)
- Xu W, "A new role for the PI3K", Akt signaling pathway in the epithelial-mesenchymal transition, 2015, pp.9, 317~324
- Grille SJ, "The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines", Cancer Res, 63, pp.2172~2178 (2003)
- Larue L, "Epithelial-mesenchymal transition in development and cancer : role of phosphatidylinositol 3’ kinase", AKT pathways, 2005, pp.24, 7443~7454
- Han Z, "Phospho Akt mediates multidrug resistance of gastric cancer cells through regulation of P-gp, Bcl-2 and Bax", J Exp Clin Cancer Res, 26, pp.261~268 (2007)
- Aberle H, "beta-catenin is a target for the ubiquitin-proteasome pathway", EMBO J, 16, pp.3797~3804 (1997)
- Osakada F, "Drug development targeting the glycogen synthase kinase-3beta(GSK-3beta)-mediated signal transduction pathway : targeting the Wnt pathway and transplantation therapy as strategies for retinal repair", J Pharmacol Sci, 109, pp.168~173 (2009)
- Wiechens N, "Nucleocytoplasmic shuttling of Axin, a negative regulator of the Wnt-betacatenin Pathway", J Biol Chem, 279, pp.5263~5267 (2004)
- Morgan RG, "Factors affecting the nuclear localization of β-catenin in normal and malignant tissue", J Cell Biochem, 115, pp.1351~1361 (2014)
- Huang L, "MUC1 oncoprotein blocks glycogen synthase kinase 3beta-mediated phosphorylation and degradation of beta-catenin", Cancer Res, 65, pp.10413~10422 (2005)
- Bautista SJ, "mTOR complex 1 controls the nuclear localization and function of glycogen synthase kinase 3β", J Biol Chem, 293, pp.14723~14739 (2018)
- He L, "Regulation of GSK3 cellular location by FRAT modulates mTORC1-dependent cell growth and sensitivity to rapamycin", Proc Natl Acad Sci U S A, 116, pp.19523~19529 (2019)
- Cong F, "Nuclear-cytoplasmic shuttling of Axin regulates subcellular localization of beta-catenin", Proc Natl Acad Sci U S A, 101, pp.2882~2887 (2004)