Abstract
Recent research has shown that Doublecortin-like kinase 1 (DCLK1) is overexpressed in different types of cancer. It has recently been described as a cancer stem cells (CSCs) marker, is associated with carcinogenesis, and positively correlates with infiltration of multiple immune cell types in some cancers. However, studies focused on assessing DCLK1 expression in HCC are limited, and the role of DCLK1 in HCC tumor immunity remains to be determined. In this study, we used a modified model of the resistant hepatocyte (MRHM) to evaluate DCLK1 expression in HCC. Furthermore, DCLK1 expression in HCC was analyzed using TIMER 2.0, UALCAN, GEPIA, GEO, and HPA web-based tools. Correlations between DCLK1 expression and clinicopathological factors in patients were analyzed using the UALCAN web-based tool. Finally, correlations between DCLK1 and immune infiltrates were investigated using the TIMER 2.0 and TISIDB web-based tools. The results showed that DCLK1 is significantly overexpressed during progression of the HCC carcinogenic process in the MRHM. DCLK1 is overexpressed in HCC according to multiple publics web-based tools, and its overexpression is associated with cancer stage. Furthermore, DCLK1 expression was correlated with infiltration levels of multiple immune cells, immunomodulatory factors, immunoinhibitors, MHC molecules, chemokines, receptors, and immune cell-specific markers. These results suggest that DCLK1 is a potential prognostic biomarker that determines cancer progression and correlates with immune cell infiltration in HCC.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
The datasets generated and/or analyzed during the present study are available on the TIMER 2.0, TISIDB, UALCAN, GEPIA, and HPA web-based tool, and are available upon request from the corresponding author.
References
Anwanwan D, Singh SK, Singh S, Saikam V, Singh R (2020) Challenges in liver cancer and possible treatment approaches. Biochim et Biophys acta Reviews cancer 1873(1):188314–188314. https://doi.org/10.1016/j.bbcan.2019.188314
Bayik D, Lathia JD (2021) Cancer stem cell-immune cell crosstalk in tumour progression. Nat Rev Cancer 21(8):526–536. https://doi.org/10.1038/s41568-021-00366-w
Broner EC, Trujillo JA, Korzinkin M, Subbannayya T, Agrawal N, Ozerov IV, Izumchenko E (2021) Doublecortin-like kinase 1 (DCLK1) is a Novel NOTCH Pathway Signaling Regulator in Head and Neck Squamous Cell Carcinoma. Front Oncol 11:677051
Cao L, Cheng H, Jiang Q, Li H, Wu Z (2020a) APEX1 is a novel diagnostic and prognostic biomarker for hepatocellular carcinoma. Aging 12(5):4573–4591. https://doi.org/10.18632/aging.102913
Cao Z, Weygant N, Chandrakesan P, Houchen CW, Peng J, Qu D (2020b) Tuft and cancer stem cell marker DCLK1: a new target to enhance anti-tumor immunity in the tumor microenvironment. Cancers (Basel) 12(12):3801. https://doi.org/10.3390/cancers12123801
Carrasco-Torres G, Monroy-Ramírez HC, Martínez-Guerra AA, Baltiérrez-Hoyos R, Romero-Tlalolini MdlÁ, Villa-Treviño S, Vásquez-Garzón VR (2017) Quercetin reverses Rat Liver Preneoplastic lesions Induced by Chemical Carcinogenesis. Oxidative Med Cell Longev 2017:4674918. https://doi.org/10.1155/2017/4674918
Chandrashekar DS, Bashel B, Balasubramanya SAH, Creighton CJ, Ponce-Rodriguez I, Chakravarthi BVSK, Varambally S (2017) UALCAN: a portal for facilitating Tumor Subgroup Gene expression and survival analyses. Neoplasia 19(8):649–658. https://doi.org/10.1016/j.neo.2017.05.002
Chidambaranathan-Reghupaty S, Fisher PB, Sarkar D (2021) Hepatocellular carcinoma (HCC): epidemiology, etiology and molecular classification. Adv Cancer Res 149:1–61. https://doi.org/10.1016/bs.acr.2020.10.001
Ding L, Yang Y, Ge Y, Lu Q, Yan Z, Chen X, Weygant N (2021) Inhibition of DCLK1 with DCLK1-IN-1 suppresses renal cell Carcinoma Invasion and Stemness and promotes cytotoxic T-Cell-mediated Anti-tumor Immunity. Cancers (Basel) 13(22):5729. https://doi.org/10.3390/cancers13225729
Fan M, Qian N, Dai G (2017) Expression and prognostic significance of doublecortin-like kinase 1 in patients with hepatocellular carcinoma. Oncol Lett 14(6):7529–7537. https://doi.org/10.3892/ol.2017.7082
Fan M, Yan H, Gou M, Qian N, Dai G (2020) Divergent expression of DCLK1 in gastrointestinal neuroendocrine tumors and primary hepatic, gallbladder, and pancreatic neuroendocrine tumors. Int J Clin Exp Pathol 13(9):2249–2258
Fukui R, Fujimoto Y, Watanabe T, Inoue N, Miyoshi Bunahiguchit Y (2020) Association between FOXP3/CD8 lymphocyte ratios and Tumor infiltrating lymphocyte levels in different breast Cancer subtypes. Anticancer Res 40:2141–2150
Gagliardi G, Goswami M, Passera R, Bellows CF (2012) DCLK1 immunoreactivity in colorectal neoplasia. Clin Exp Gastroenterol 5:35–42. https://doi.org/10.2147/CEG.S30281
Gao T, Wang M, Xu L, Wen T, Liu J, An G (2016) DCLK1 is up-regulated and associated with metastasis and prognosis in colorectal cancer. J Cancer Res Clin Oncol 142(10):2131–2140. https://doi.org/10.1007/s00432-016-2218-0
Granito A, Muratori L, Lalanne C, Quarneti C, Ferri S, Guidi M, Muratori P (2021) Hepatocellular carcinoma in viral and autoimmune liver diseases: role of CD4 + CD25 + Foxp3 + regulatory T cells in the immune microenvironment. World J Gastroenterol 27(22):2994–3009. https://doi.org/10.3748/wjg.v27.i22.2994
Guo Y, Yang J, Ren K, Tian X, Gao H, Tian X, Kan Q (2022) The heterogeneity of immune cell infiltration landscape and its immunotherapeutic implications in hepatocellular carcinoma. Front Immunol 13:861525
Khalaf K, Hana D, Chou JT-T, Singh C, Mackiewicz A, Kaczmarek M (2021) Aspects of the Tumor Microenvironment involved in Immune Resistance and Drug Resistance. Front Immunol 12:656364–656364. https://doi.org/10.3389/fimmu.2021.656364
Lee TK, Guan XY, Ma S (2022) Cancer stem cells in hepatocellular carcinoma - from origin to clinical implications. Nat Rev Gastroenterol Hepatol 19(1):26–44. https://doi.org/10.1038/s41575-021-00508-3
Leffers N, Gooden MJM, de Jong RA, Hoogeboom B-N, ten Hoor KA, Hollema H, Nijman HW (2009) Prognostic significance of tumor-infiltrating T-lymphocytes in primary and metastatic lesions of advanced stage ovarian cancer. Cancer Immunol Immunother 58(3):449–459. https://doi.org/10.1007/s00262-008-0583-5
Li J, Wang Y, Ge J, Li W, Yin L, Zhao Z, Wang H (2018) Doublecortin-like kinase 1 (DCLK1) regulates B Cell-Specific Moloney murine leukemia virus insertion site 1 (Bmi-1) and is Associated with Metastasis and Prognosis in Pancreatic Cancer. Cell Physiol Biochem 51(1):262–277. https://doi.org/10.1159/000495228
Li T, Fu J, Zeng Z, Cohen D, Li J, Chen Q, Liu XS (2020) TIMER2.0 for analysis of tumor-infiltrating immune cells. Nucleic Acids Res 48(W1):W509–w514. https://doi.org/10.1093/nar/gkaa407
Lim H-Y, Sohn I, Deng S, Lee J, Jung SH, Mao M, Xu J, Wang K, Shi S, Joh JW, Choi YL, Park C-K (2013) Prediction of Disease-free Survival in Hepatocellular Carcinoma by Gene Expression Profiling. Ann Surg Oncol 20(12):3747–3753. https://doi.org/10.1245/s10434-013-3070-y
Liu CY, Chen KF, Chen PJ (2015) Treatment of Liver Cancer. Cold Spring Harb Perspect Med 5(9):a021535. https://doi.org/10.1101/cshperspect.a021535
Liu X, Long X, Liu W, Zhao Y, Hayashi T, Yamato M, Ikejima T (2018) Type I collagen induces mesenchymal cell differentiation into myofibroblasts through YAP-induced TGF-β1 activation. Biochimie 150:110–130. https://doi.org/10.1016/j.biochi.2018.05.005
Liu H, Wen T, Zhou Y, Fan X, Du T, Gao T, An G (2019) DCLK1 plays a metastatic-promoting role in human breast cancer cells. Biomed Res Int 2019:1061979. https://doi.org/10.1155/2019/1061979
Liu Y-C, Yeh C-T, Lin K-H (2020) Cancer Stem Cell functions in Hepatocellular Carcinoma and Comprehensive therapeutic strategies. Cells 9(6):1331. https://doi.org/10.3390/cells9061331
Llovet JM, Kelley RK, Villanueva A, Singal AG, Pikarsky E, Roayaie S, Finn RS (2021) Hepatocellular carcinoma. Nat Rev Dis Primers 7(1):6. https://doi.org/10.1038/s41572-020-00240-3
Lv Y, Song G, Wang R, Di L, Wang J (2017) Doublecortin-like kinase 1 is a novel biomarker for prognosis and regulates growth and metastasis in basal-like breast cancer. Biomed Pharmacother 88:1198–1205. https://doi.org/10.1016/j.biopha.2017.01.082
Mahmoud SM, Paish EC, Powe DG, Macmillan RD, Lee AH, Ellis IO, Green AR (2011) An evaluation of the clinical significance of FOXP3 + infiltrating cells in human breast cancer. Breast Cancer Res Treat 127(1):99–108. https://doi.org/10.1007/s10549-010-0987-8
Ogunwobi OO, Harricharran T, Huaman J, Galuza A, Odumuwagun O, Tan Y, Nguyen MT (2019) Mechanisms of hepatocellular carcinoma progression. World J Gastroenterol 25(19):2279–2293. https://doi.org/10.3748/wjg.v25.i19.2279
Panneerselvam J, Mohandoss P, Patel R, Gillan H, Li M, Kumar K, Chandrakesan P (2020) DCLK1 regulates Tumor Stemness and Cisplatin Resistance in Non-small Cell Lung Cancer via ABCD-Member-4. Mol Therapy - Oncolytics 18:24–36. https://doi.org/10.1016/j.omto.2020.05.012
Ren Z, Ma X, Duan Z, Chen X (2020) Diagnosis, therapy, and prognosis for hepatocellular carcinoma. Anal Cell Pathol 2020:8157406. https://doi.org/10.1155/2020/8157406
Ru B, Wong CN, Tong Y, Zhong JY, Zhong SSW, Wu WC, Zhang J (2019) TISIDB: an integrated repository portal for tumor-immune system interactions. Bioinformatics 35(20):4200–4202. https://doi.org/10.1093/bioinformatics/btz210
Sánchez PS, Rigual MD, Djouder N (2021) Inflammatory and non-inflammatory mechanisms Controlling Cirrhosis Development. Cancers (Basel) 13(20):5045
Shang B, Liu Y, Jiang SJ, Liu Y (2015) Prognostic value of tumor-infiltrating FoxP3 + regulatory T cells in cancers: a systematic review and meta-analysis. Sci Rep 5:15179. https://doi.org/10.1038/srep15179
Shimada S, Mogushi K, Akiyama Y, Furuyama T, Watanabe S, Ogura T, Ogawa K, Ono H, Mitsunori Y, Ban D, Kudo A, Arii S, Tanabe M, Wands JR, Tanaka S (2019) Comprehensive molecular and immunological characterization of hepatocellular carcinoma. EBioMedicine 40:457–470. https://doi.org/10.1016/j.ebiom.2018.12.058
Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F (2021) Global Cancer statistics 2020: GLOBOCAN estimates of incidence and Mortality Worldwide for 36 cancers in 185 countries. CA Cancer J Clin 71(3):209–249. https://doi.org/10.3322/caac.21660
Sureban SM, Madhoun MF, May R, Qu D, Ali N, Fazili J, Houchen CW (2015) Plasma DCLK1 is a marker of hepatocellular carcinoma (HCC): Targeting DCLK1 prevents HCC tumor xenograft growth via a microRNA-dependent mechanism. Oncotarget 6(35):37200–37215. https://doi.org/10.18632/oncotarget.5808
Tang Z, Li C, Kang B, Gao G, Li C, Zhang Z (2017) GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Res 45(W1):W98–W102. https://doi.org/10.1093/nar/gkx247
Uhlén M, Fagerberg L, Hallström BM, Lindskog C, Oksvold P, Mardinoglu A, Pontén F (2015) Proteomics. Tissue-based map of the human proteome. Science 347(6220):1260419. https://doi.org/10.1126/science.1260419
Vásquez-Garzón VR, Macias-Pérez JR, Jiménez-García MN, Villegas V, Fattel-Fazenta S, Villa-Treviño S (2013) The chemopreventive capacity of quercetin to induce programmed cell death in hepatocarcinogenesis. Toxicol Pathol 41(6):857–865. https://doi.org/10.1177/0192623312467522
Vásquez-Garzón VR, Beltrán-Ramírez O, Salcido-Neyoy ME, Cervante-Anaya N, Villa-Treviño S (2015) Analysis of gene expression profiles as a tool to uncover tumor markers of liver cancer progression in a rat model. Biomed Rep 3(2):167–172. https://doi.org/10.3892/br.2014.411
Vijai M, Baba M, Ramalingam S, Thiyagaraj A (2021) DCLK1 and its interaction partners: an effective therapeutic target for colorectal cancer. Oncol Lett 22(6):850–850. https://doi.org/10.3892/ol.2021.13111
Wang SM, Ooi LL, Hui KM (2007) Identification and validation of a novel gene signature associated with the recurrence of human hepatocellular carcinoma. Clin Cancer Res 13(21):6275–6283. https://doi.org/10.1158/1078-0432.CCR-06-2236
West NR, Kost SE, Martin SD, Milne K, Deleeuw RJ, Nelson BH, Watson PH (2013) Tumour-infiltrating FOXP3(+) lymphocytes are associated with cytotoxic immune responses and good clinical outcome in oestrogen receptor-negative breast cancer. Br J Cancer 108(1):155–162. https://doi.org/10.1038/bjc.2012.524
Weygant N, Qu D, May R, Tierney RM, Berry WL, Zhao L, Houchen CW (2014) DCLK1 is a broadly dysregulated target against epithelial-mesenchymal transition, focal adhesion, and stemness in clear cell renal carcinoma. Oncotarget 6(4):2193
Wu X, Qu D, Weygant N, Peng J, Houchen CW (2020) Cancer stem cell marker DCLK1 correlates with tumorigenic Immune infiltrates in the Colon and gastric adenocarcinoma microenvironments. Cancers (Basel) 12(2):274. https://doi.org/10.3390/cancers12020274
Yan R, Li J, Zhou Y, Yao L, Sun R, Xu Y, An G (2020) Inhibition of DCLK1 down-regulates PD-L1 expression through Hippo pathway in human pancreatic cancer. Life Sci 241:117150. https://doi.org/10.1016/j.lfs.2019.117150
Yan R, Li J, Xiao Z, Fan X, Liu H, Xu Y, Ge Y (2023) DCLK1 suppresses tumor-specific cytotoxic T lymphocyte function through recruitment of MDSCs via the CXCL1-CXCR2 Axis. Cell Mol Gastroenterol Hepatol 15(2):463–485. https://doi.org/10.1016/j.jcmgh.2022.10.013
Yang JD, Hainaut P, Gores GJ, Amadou A, Plymoth A, Roberts LR (2019) A global view of hepatocellular carcinoma: trends, risk, prevention and management. Nat Rev Gastroenterol Hepatol 16(10):589–604. https://doi.org/10.1038/s41575-019-0186-y
Funding
This research was funded by the Consejo Nacional de Ciencia y Tecnología CONACyT, grant No. CF2019-53358 to Verónica Rocío Vásquez-Garzón, and Saúl Villa-Treviño.
Author information
Authors and Affiliations
Contributions
All authors contributed to the conception and design of the study. Material preparation, data collection, and data analysis were performed by Juan Manuel Velázquez-Enríquez, Renata Cerna, Olga Beltrán-Ramírez, Carolina Piña-Vázquez, Saúl Villa-Treviño, and Verónica Rocío Vásquez-Garzón. The first draft of the manuscript was written by Juan Manuel Velázquez-Enríquez, Renata Cerna. Critical reading and correction of the manuscript was performed by Juan Manuel Velázquez-Enríquez, Renata Cerna, Olga Beltrán-Ramírez, Carolina Piña-Vázquez, Saúl Villa-Treviño, and Verónica Rocío Vásquez-Garzón. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Ethical approval
All experimental animal studies were performed with the approval of the Institutional Animal Care and Use Committee (IACUC) of CINVESTAV-IPN, protocol 0168 − 15.
Informed Consent
Not applicable.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Velázquez-Enríquez, J.M., Cerna, R., Beltrán-Ramírez, O. et al. DCLK1 is Overexpressed and Associated with Immune Cell Infiltration in Hepatocellular Carcinoma. Biochem Genet (2024). https://doi.org/10.1007/s10528-024-10667-y
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10528-024-10667-y