Summary
Acquired resistance is a significant hindrance to clinical application of lenvatinib in unresectable hepatocellular carcinoma (HCC). Further in-depth investigation of resistance mechanisms can help to develop additional therapeutic strategies to overcome or delay resistance. In our study, two lenvatinib-resistant (LR) HCC cell lines were established by treatment with gradient increasing concentration of lenvatinib, named Hep3B-LR and HepG2-LR. Interestingly, continuous lenvatinib treatment reinforced epithelial-mesenchymal transition (EMT), cell migration, and cell invasion. Gene set enrichment analysis (GSEA) enrichment analysis of RNA-sequencing from Hep3B-LR and corresponding parental cells revealed that activation of Wnt signaling pathway was involved in this adaptive process. Active β-catenin and its downstream target lymphoid enhancer binding factor 1 (LEF1) were significantly elevated in LR HCC cells, which promoted lenvatinib resistance through mediating EMT-related genes. Data analysis based on Gene Expression Omnibus (GEO) and the Cancer Genome Atlas Program (TCGA) databases suggests that LEF1, as a key regulator of EMT, was a novel molecular target linked to lenvatinib resistance and poor prognosis in HCC. Using a small-molecule specific inhibitor ICG001 and knocking down LEF1 showed that targeting LEF1 restored the sensitivity of LR HCC cells to lenvatinib. Our results uncover upregulation of LEF1 confers lenvatinib resistance by facilitating EMT, cell migration, and invasion of LR HCC cells, indicating that LEF1 is a novel therapeutic target for overcoming acquired lenvatinib resistance.
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References
Siegel RL, Miller KD, Fuchs HE, Jemal A (2021) Cancer statistics, 2021. CA Cancer J Clin 71(1):7–33. https://doi.org/10.3322/caac.21654
Llovet JM, Pinyol R, Kelley RK, El-Khoueiry A, Reeves HL, Wang XW et al (2022) Molecular pathogenesis and systemic therapies for hepatocellular carcinoma. Nat Cancer 3(4):386–401. https://doi.org/10.1038/s43018-022-00357-2
Llovet JM, Kelley RK, Villanueva A, Singal AG, Pikarsky E, Roayaie S et al (2021) Hepatocellular carcinoma. Nat Rev Dis Primers 7(1):6. https://doi.org/10.1038/s41572-020-00240-3
Sangro B, Sarobe P, Hervas-Stubbs S, Melero I (2021) Advances in immunotherapy for hepatocellular carcinoma. Nat Rev Gastroenterol Hepatol 18(8):525–543. https://doi.org/10.1038/s41575-021-00438-0
Matsui J, Yamamoto Y, Funahashi Y, Tsuruoka A, Watanabe T, Wakabayashi T et al (2008) E7080, a novel inhibitor that targets multiple kinases, has potent antitumor activities against stem cell factor producing human small cell lung cancer H146, based on angiogenesis inhibition. Int J Cancer 122(3):664–671. https://doi.org/10.1002/ijc.23131
Matsui J, Funahashi Y, Uenaka T, Watanabe T, Tsuruoka A, Asada M (2008) Multi-kinase inhibitor E7080 suppresses lymph node and lung metastases of human mammary breast tumor MDA-MB-231 via inhibition of vascular endothelial growth factor-receptor (VEGF-R) 2 and VEGF-R3 kinase. Clin Cancer Res 14(17):5459–5465. https://doi.org/10.1158/1078-0432.CCR-07-5270
Wang J, Yu H, Dong W, Zhang C, Hu M, Ma W et al (2023) N6-methyladenosine-mediated up-regulation of FZD10 regulates liver cancer stem cells’ properties and lenvatinib resistance through WNT/beta-catenin and hippo signaling pathways. Gastroenterology 164(6):990–1005. https://doi.org/10.1053/j.gastro.2023.01.041
Tian Y, Lei Y, Fu Y, Sun H, Wang J, Xia F (2022) Molecular mechanisms of resistance to tyrosine kinase inhibitors associated with hepatocellular carcinoma. Curr Cancer Drug Targets 22(6):454–462. https://doi.org/10.2174/1568009622666220330151725
Tang W, Chen Z, Zhang W, Cheng Y, Zhang B, Wu F et al (2020) The mechanisms of sorafenib resistance in hepatocellular carcinoma: theoretical basis and therapeutic aspects. Signal Transduct Target Ther 5(1):87. https://doi.org/10.1038/s41392-020-0187-x
da Fonseca LG, Reig M, Bruix J (2020) Tyrosine kinase inhibitors and hepatocellular carcinoma. Clin Liver Dis 24(4):719–737. https://doi.org/10.1016/j.cld.2020.07.012
Pan J, Zhang M, Dong L, Ji S, Zhang J, Zhang S et al (2023) Genome-Scale CRISPR screen identifies LAPTM5 driving lenvatinib resistance in hepatocellular carcinoma. Autophagy 19(4):1184–1198. https://doi.org/10.1080/15548627.2022.2117893
Ao J, Chiba T, Shibata S, Kurosugi A, Qiang N, Ma Y et al (2021) Acquisition of mesenchymal-like phenotypes and overproduction of angiogenic factors in lenvatinib-resistant hepatocellular carcinoma cells. Biochem Biophys Res Commun 549:171–178. https://doi.org/10.1016/j.bbrc.2021.02.097
Hu B, Zou T, Qin W, Shen X, Su Y, Li J et al (2022) Inhibition of EGFR overcomes acquired lenvatinib resistance driven by STAT3-ABCB1 signaling in hepatocellular carcinoma. Cancer Res 82(20):3845–3857. https://doi.org/10.1158/0008-5472.CAN-21-4140
Huang S, Ma Z, Zhou Q, Wang A, Gong Y, Li Z et al (2022) Genome-wide CRISPR/Cas9 library screening identified that DUSP4 deficiency induces lenvatinib resistance in hepatocellular carcinoma. Int J Biol Sci 18(11):4357–4371. https://doi.org/10.7150/ijbs.69969
Lu Y, Shen H, Huang W, He S, Chen J, Zhang D et al (2021) Genome-scale CRISPR-Cas9 knockout screening in hepatocellular carcinoma with lenvatinib resistance. Cell Death Discov 7(1):359. https://doi.org/10.1038/s41420-021-00747-y
He X, Hikiba Y, Suzuki Y, Nakamori Y, Kanemaru Y, Sugimori M et al (2022) EGFR inhibition reverses resistance to lenvatinib in hepatocellular carcinoma cells. Sci Rep 12(1):8007. https://doi.org/10.1038/s41598-022-12076-w
Huang M, Long J, Yao Z, Zhao Y, Zhao Y, Liao J et al (2023) METTL1-mediated m7G tRNA modification promotes lenvatinib resistance in hepatocellular carcinoma. Cancer Res 83(1):89–102. https://doi.org/10.1158/0008-5472.CAN-22-0963
Duan A, Li H, Yu W, Zhang Y, Yin L (2022) Long noncoding RNA XIST promotes resistance to lenvatinib in hepatocellular carcinoma cells via epigenetic inhibition of NOD2. J Oncol 2022:4537343. https://doi.org/10.1155/2022/4537343
Ozvegy-Laczka C, Cserepes J, Elkind NB, Sarkadi B (2005) Tyrosine kinase inhibitor resistance in cancer: role of ABC multidrug transporters. Drug Resist Updat 8(1–2):15–26. https://doi.org/10.1016/j.drup.2005.02.002
Loh CY, Chai JY, Tang TF, Wong WF, Sethi G, Shanmugam MK et al (2019) The E-cadherin and N-cadherin switch in epithelial-to-mesenchymal transition: Signaling, therapeutic implications, and challenges. Cells 8(10). https://doi.org/10.3390/cells8101118
Emami KH, Nguyen C, Ma H, Kim DH, Jeong KW, Eguchi M et al (2004) A small molecule inhibitor of beta-catenin/CREB-binding protein transcription [corrected]. Proc Natl Acad Sci U S A 101(34):12682–12687. https://doi.org/10.1073/pnas.0404875101
Henderson WR, Jr., Chi E Y, Ye X, Nguyen C, Tien Y T, Zhou B, et al (2010) Inhibition of Wnt/beta-catenin/CREB binding protein (CBP) signaling reverses pulmonary fibrosis. Proc Natl Acad Sci U S A 107(32):14309–14314. https://doi.org/10.1073/pnas.1001520107
Chen CL, Tsai YS, Huang YH, Liang YJ, Sun YY, Su CW et al (2018) Lymphoid enhancer factor 1 contributes to hepatocellular carcinoma progression through transcriptional regulation of epithelial-mesenchymal transition regulators and stemness genes. Hepatol Commun 2(11):1392–1407. https://doi.org/10.1002/hep4.1229
Kobayashi W, Ozawa M (2013) The transcription factor LEF-1 induces an epithelial-mesenchymal transition in MDCK cells independent of beta-catenin. Biochem Biophys Res Commun 442(1–2):133–138. https://doi.org/10.1016/j.bbrc.2013.11.031
Basu S, Cheriyamundath S, Ben-Ze'ev A (2018) Cell-cell adhesion: linking Wnt/beta-catenin signaling with partial EMT and stemness traits in tumorigenesis. F1000Res 7. https://doi.org/10.12688/f1000research.15782.1
Shibue T, Weinberg RA (2017) EMT, CSCs, and drug resistance: the mechanistic link and clinical implications. Nat Rev Clin Oncol 14(10):611–629. https://doi.org/10.1038/nrclinonc.2017.44
Dart A (2023) EMT in chemoresistance. Nat Rev Cancer 23(6):349. https://doi.org/10.1038/s41568-023-00581-7
Clevers H (2006) Wnt/beta-catenin signaling in development and disease. Cell 127(3):469–480. https://doi.org/10.1016/j.cell.2006.10.018
Lambertini E, Franceschetti T, Torreggiani E, Penolazzi L, Pastore A, Pelucchi S et al (2010) SLUG: a new target of lymphoid enhancer factor-1 in human osteoblasts. BMC Mol Biol 11:13. https://doi.org/10.1186/1471-2199-11-13
Santiago L, Daniels G, Wang D, Deng FM, Lee P (2017) Wnt signaling pathway protein LEF1 in cancer, as a biomarker for prognosis and a target for treatment. Am J Cancer Res 7(6):1389–1406
Funding
Our research was supported by the National Natural Science Foundation of China (NO. 81972664) and Natural Science Foundation of Jiangsu Province (No. BK20231419).
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Xinxiu Li carried out the in vitro experiments and drafted the manuscript. Hongmeng Su, Luyu Zhao, and Jinghan Sun implemented bioinformatics analysis. Shihui Shu and Wenqing Tang performed cell culture and revised the manuscript. Hong Fan designed the project. All authors read and approved the final manuscript.
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Li, X., Su, H., Tang, W. et al. Targeting LEF1-mediated epithelial-mesenchymal transition reverses lenvatinib resistance in hepatocellular carcinoma. Invest New Drugs 42, 185–195 (2024). https://doi.org/10.1007/s10637-024-01426-2
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DOI: https://doi.org/10.1007/s10637-024-01426-2