KIF2C: a novel link between Wnt/β-catenin and mTORC1 signaling in the pathogenesis of hepatocellular carcinoma

Abstract

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is the fourth-leading cause of cancer-related deaths worldwide. HCC is refractory to many standard cancer treatments and the prognosis is often poor, highlighting a pressing need to identify biomarkers of aggressiveness and potential targets for future treatments. Kinesin family member 2C (KIF2C) is reported to be highly expressed in several human tumors. Nevertheless, the molecular mechanisms underlying the role of KIF2C in tumor development and progression have not been investigated. In this study, we found that KIF2C expression was significantly upregulated in HCC, and that KIF2C up-regulation was associated with a poor prognosis. Utilizing both gain and loss of function assays, we showed that KIF2C promoted HCC cell proliferation, migration, invasion, and metastasis both in vitro and in vivo. Mechanistically, we identified TBC1D7 as a binding partner of KIF2C, and this interaction disrupts the formation of the TSC complex, resulting in the enhancement of mammalian target of rapamycin complex1 (mTORC1) signal transduction. Additionally, we found that KIF2C is a direct target of the Wnt/β-catenin pathway, and acts as a key factor in mediating the crosstalk between Wnt/β-catenin and mTORC1 signaling. Thus, the results of our study establish a link between Wnt/β-catenin and mTORC1 signaling, which highlights the potential of KIF2C as a therapeutic target for the treatment of HCC.

Keywords

KIF2C / HCC / TBC1D7 / mTORC1 signaling / Wnt/β-catenin signaling

References

[1] Ally A, Balasundaram M, Carlsen R, Chuah E, Clarke A, Dhalla N, Holt RA, Jones SJM, Lee D, Ma Y (2017) Comprehensive and integrative genomic characterization of hepatocellular carcinoma. Cell 169:1327

[2] Bhat M, Sonenberg N, Gores GJ (2013) The mTOR pathway in hepatic malignancies. Hepatology 58:810–818

[3] Bie L, Zhao G, Wang YP, Zhang B (2012) Kinesin family member 2C (KIF2C/MCAK) is a novel marker for prognosis in human gliomas. Clin Neurol Neurosurg 114:356–360

[4] Braun A, Dang K, Buslig F, Baird MA, Davidson MW, Waterman CM, Myers KA (2014) Rac1 and Aurora A regulate MCAK to polarize microtubule growth in migrating endothelial cells. J Cell Biol 206:97–112

[5] Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68:394–424

[6] Capo-Chichi JM, Tcherkezian J, Hamdan FF, Decarie JC, Dobrzeniecka S, Patry L, Nadon MA, Mucha BE, Major P, Shevell M (2013) Disruption of TBC1D7, a subunit of the TSC1-TSC2 protein complex, in intellectual disability and megalencephaly. J Med Genet 50:740–744

[7] Chang CY, Lin SC, Su WH, Ho CM, Jou YS (2012) Somatic LMCD1 mutations promoted cell migration and tumor metastasis in hepatocellular carcinoma. Oncogene 31:2640–2652

[8] Chen JX, Rajasekaran M,Xia HP, Zhang XQ, Kong SN, Sekar K, Seshachalam VP, Deivasigamani A, Goh BKP, Ooi LL (2016) The microtubule-associated protein PRC1 promotes early recurrence of hepatocellular carcinoma in association with the Wnt/beta-catenin signalling pathway. Gut 65:1522–1534

[9] Chen JL, Li S, Zhou S, Cao SJ, Lou Y, Shen HY, Yin J, Li G (2017) Kinesin superfamily protein expression and its association with progression and prognosis in hepatocellular carcinoma. J Cancer Res Ther 13:651–659

[10] Choi YJ, Park YJ, Park JY, Jeong HO, Kim DH, Ha YM, Kim JM, Song YM, Heo HS, Yu BP (2012) Inhibitory effect of mTOR activator MHY1485 on autophagy: suppression of lysosomal fusion. PLoS ONE 7:e43418

[11] Clevers H, Nusse R (2012) Wnt/beta-catenin signaling and disease. Cell 149:1192–1205

[12] Dibble CC, Elis W, Menon S, Qin W, Klekota J, Asara JM, Finan PM, Kwiatkowski DJ, Murphy LO, Manning BD (2012) TBC1D7 Is a third subunit of the TSC1-TSC2 complex upstream of mTORC1. Mol Cell 47:535–546

[13] Eichenlaub-Ritter U (2015) Microtubule dynamics and tumor invasion involving MCAK. Cell Cycle 14:3353

[14] El-Serag HB, Mason AC (1999) Rising incidence of hepatocellular carcinoma in the United States. New Engl J Med 340:745–750

[15] Gan HZ, Lin L, Hu NJ, Yang Y, Gao Y, Pei Y, Chen K, Sun BT (2019) KIF2C exerts an oncogenic role in nonsmall cell lung cancer and is negatively regulated by miR-325-3p. Cell Biochem Funct 37:424–431

[16] Gonsalves FC, Klein K, Carson BB, Katz S,Ekas LA, Evans S, Nagourney R, Cardozo T, Brown AM, DasGupta R (2011) An RNAi-based chemical genetic screen identifies three smallmolecule inhibitors of the Wnt/wingless signaling pathway. Proc Natl Acad Sci USA 108:5954–5963

[17] Grandy D, Shan JF, Zhang XX, Rao S, Akunuru S, Li HY, Zhang YH, Alpatov I,Zhang XA, Lang RA (2009) Discovery and characterization of a small molecule inhibitor of the PDZ domain of dishevelled. J Biol Chem 284:16256–16263

[18] Gulhati P, Bowen KA, Liu J,Stevens PD, Rychahou PG, Chen M, Lee EY, Weiss HL,O’Connor KL, Gao T (2011) mTORC1 and mTORC2 regulate EMT, motility, and metastasis of colorectal cancer via RhoA and Rac1 signaling pathways. Cancer Res 71:3246–3256

[19] Guo P, Ma X, Zhao W, Huai W, Li T, Qiu Y, Zhang Y, Han L (2018) TRIM31 is upregulated in hepatocellular carcinoma and promotes disease progression by inducing ubiquitination of TSC1-TSC2 complex. Oncogene 37:478–488

[20] Harris TE, Lawrence JC Jr (2003) TOR signaling. Sci STKE 2003:15

[21] Hedgepeth CM, Conrad LJ, Zhang J, Huang HC, Lee VM, Klein PS (1997) Activation of the Wnt signaling pathway: a molecular mechanism for lithium action. Dev Biol 185:82–91

[22] Hirokawa N, Noda Y, Tanaka Y, Niwa S (2009) Kinesin superfamily motor proteins and intracellular transport. Nat Rev Mol Cell Biol 10:682–696

[23] Honnappa S, Gouveia SM, Weisbrich A, Damberger FF, Bhavesh NS, Jawhari H, Grigoriev I,van Rijssel FJA, Buey RM, Lawera A (2009) An EB1-binding motif acts as a microtubule tip localization signal. Cell 138:366–376

[24] Hsieh AC, Liu Y, Edlind MP, Ingolia NT, Janes MR, Sher A, Shi EY, Stumpf CR, Christensen C, Bonham MJ (2012) The translational landscape of mTOR signalling steers cancer initiation and metastasis. Nature 485:55–61

[25] Huang SMA, Mishina YM, Liu SM, Cheung A, Stegmeier F, Michaud GA, Charlat O, Wiellette E, Zhang Y, Wiessner S (2009) Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling. Nature 461:614–620

[26] Huang Y, Wang H, Lian Y, Wu X, Zhou L, Wang J, Deng M, Huang Y (2018) Upregulation of kinesin family member 4A enhanced cell proliferation via activation of Akt signaling and predicted a poor prognosis in hepatocellular carcinoma. Cell Death Dis 9:141

[27] Ince N, Wands JR (1999) The increasing incidence of hepatocellular carcinoma. New Engl J Med 340:798–799

[28] Inoki K, Corradetti MN, Guan KL (2005) Dysregulation of the TSCmTOR pathway in human disease. Nat Genet 37:19–24

[29] Inoki K,Ouyang H, Zhu TQ, Lindvall C, Wang Y, Zhang XJ, Yang Q, Bennett C, Harada Y, Stankunas K (2006) TSC2 integrates Wnt and energy signals via a coordinated phosphorylation by AMPK and GSK3 to regulate cell growth. Cell 126:955–968

[30] Ishikawa K, Kamohara Y, Tanaka F, Haraguchi N, Mimori K, Inoue H, Mori M (2008) Mitotic centromere-associated kinesin is a novel marker for prognosis and lymph node metastasis in colorectal cancer. Br J Cancer 98:1824–1829

[31] Jacinto E, Hall MN (2003) Tor signalling in bugs, brain and brawn. Nat Rev Mol Cell Biol 4:117–126

[32] Khalaf AM, Fuentes D, Morshid AI, Burke MR, Kaseb AO, Hassan M, Hazle JD, Elsayes KM (2018) Role of Wnt/beta-catenin signaling in hepatocellular carcinoma, pathogenesis, and clinical significance. J Hepatocell Carcinoma 5:61–73

[33] Lachenmayer A, Alsinet C, Savic R, Cabellos L, Toffanin S, Hoshida Y, Villanueva A, Minguez B, Newell P, Tsai HW (2012) Wntpathway activation in two molecular classes of hepatocellular carcinoma and experimental modulation by sorafenib. Clin Cancer Res 18:4997–5007

[34] Lee T, Langford KJ, Askham JM, Bruning-Richardson A, Morrison EE (2008) MCAK associates with EB1. Oncogene 27:2494–2500

[35] Li H, Shen S, Chen X, Ren Z, Li Z, Yu Z (2019a) miR-450b-5p loss mediated KIF26B activation promoted hepatocellular carcinoma progression by activating PI3K/AKT pathway. Cancer Cell Int 19:205

[36] Li Q, Pan XX, Zhu DM, Deng ZM, Jiang RQ, Wang XH (2019b) Circular RNA MAT2B promotes glycolysis and malignancy of hepatocellular carcinoma through the miR-338-3p/PKM2 axis under hypoxic stress. Hepatology 70:1298–1316

[37] Li Q, Qiu J, Yang H, Sun G, Hu Y, Zhu D, Deng Z, Wang X, Tang J, Jiang R (2020) Kinesin family member 15 promotes cancer stem cell phenotype and malignancy via reactive oxygen species imbalance in hepatocellular carcinoma. Cancer Lett 482:112–125

[38] Luo W, Liao M, Liao Y, Chen X, Huang C,Fan J, Liao W (2018) The role of kinesin KIF18A in the invasion and metastasis of hepatocellular carcinoma. World J Surg Oncol 16:36

[39] Mamane Y, Petroulakis E, Rong LW, Yoshida K, Ler LW, Sonenberg N (2004) eIF4E—from translation to transformation. Oncogene 23:3172–3179

[40] Matter MS, Decaens T, Andersen JB, Thorgeirsson SS (2014) Targeting the mTOR pathway in hepatocellular carcinoma: current state and future trends . J Hepatol 60:855–865

[41] Moore AT, Rankin KE, von Dassow G, Peris L, Wagenbach M, Ovechkina Y, Andrieux A, Job D, Wordeman L (2005) MCAK associates with the tips of polymerizing microtubules. J Cell Biol 169:391–397

[42] Nakamura Y, Tanaka F, Haraguchi N, Mimori K, Matsumoto T, Inoue H, Yanaga K, Mori M (2007) Clinicopathological and biological significance of mitotic centromere-associated kinesin overexpression in human gastric cancer. Br J Cancer 97:543–549

[43] Njei B, Rotman Y, Ditah I, Lim JK (2015) Emerging trends in hepatocellular carcinoma incidence and mortality. Hepatology 61:191–199

[44] Pez F,Lopez A, Kim M,Wands JR, de Fromentel CC, Merle P (2013) Wnt signaling and hepatocarcinogenesis: molecular targets for the development of innovative anticancer drugs. J Hepatol 59:1107–1117

[45] Ritter A, Sanhaji M, Friemel A, Roth S, Rolle U, Louwen F, Yuan JP (2015) Functional analysis of phosphorylation of the mitotic centromere-associated kinesin by Aurora B kinase in human tumor cells. Cell Cycle 14:3755–3767

[46] Ritter A, Kreis NN, Louwen F, Wordeman L, Yuan JP (2016) Molecular insight into the regulation and function of MCAK. Crit Rev Biochem Mol 51:228–245

[47] Roessler S, Jia HL, Budhu A, Forgues M, Ye QH, Lee JS, Thorgeirsson SS, Sun ZT, Tang ZY, Qin LX (2010) A unique metastasis gene signature enables prediction of tumor relapse in early-stage hepatocellular carcinoma patients. Cancer Res 70:10202–10212

[48] Saucedo LJ, Gao XS, Chiarelli DA, Li L, Pan D, Edgar BA (2003) Rheb promotes cell growth as a component of the insulin/TOR signalling network. Nat Cell Biol 5:566–571

[49] Saxton RA, Sabatini DM (2017) mTOR signaling in growth, metabolism, and disease. Cell 168:960–976

[50] Shimo A, Tanikawa C, Nishidate T, Lin ML, Matsuda K, Park JH, Ueki T, Ohta T, Hirata K, Fukuda M (2008) Involvement of kinesin family member 2C/mitotic centromere-associated kinesin overexpression in mammary carcinogenesis. Cancer Sci 99:62–70

[51] Tanaka S, Arii S (2012) Molecular targeted therapies in hepatocellular carcinoma. Semin Oncol 39:486–492

[52] Tanenbaum ME, Medema RH, Akhmanova A (2011) Regulation of localization and activity of the microtubule depolymerase MCAK. Bioarchitecture 1:80–87

[53] Teng K, Wei S, Zhang C, Chen JW, Chen JB, Xiao KH, Liu J, Dai MM, Guan XY, Yun JP (2019) KIFC1 is activated by TCF-4 and promotes hepatocellular carcinoma pathogenesis by regulating HMGA1 transcriptional activity. J Exp Clin Cancer Res 38:329

[54] Villanueva A, Chiang DY, Newell P, Peix J, Thung S, Alsinet C,Tovar V, Roayaie S, Minguez B, Sole M (2008) Pivotal role of mTOR signaling in hepatocellular carcinoma. Gastroenterology 135:1972–1983

[55] Zhang Y, Gao XS, Saucedo LJ, Ru BG, Edgar BA, Pan DJ (2003) Rheb is a direct target of the tuberous sclerosis tumour suppressor proteins. Nat Cell Biol 5:578–581

[56] Zhang GP, Shen SL, Yu Y, Yue X, Hu WJ, Li SQ (2020) Kinesin family member 2C aggravates the progression of hepatocellular carcinoma and interacts with competing endogenous RNA. J Cell Biochem.https://doi.org/10.1002/jcb.29665

[57] Zhu W, Cai MY, Tong ZT, Dong SS, Mai SJ, Liao YJ, Bian XW, Lin MC, Kung HF, Zeng YX (2012) Overexpression of EIF5A2 promotes colorectal carcinoma cell aggressiveness by upregulating MTA1 through C-myc to induce epithelial-mesenchymaltransition. Gut 61:562–575

RIGHTS & PERMISSIONS

2020 The Author(s)