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Journal of Experimental & Clinical Cancer Research

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Open Access 01-12-2019 | Hepatocellular Carcinoma | Research

Musashi2 contributes to the maintenance of CD44v6+ liver cancer stem cells via notch1 signaling pathway

Authors: Xiju Wang, Ronghua Wang, Shuya Bai, Si Xiong, Yawen Li, Man Liu, Zhenxiong Zhao, Yun Wang, Yuchong Zhao, Wei Chen, Timothy R. Billiar, Bin Cheng

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2019

Abstract

Background

Liver cancer stem cells (LCSCs) contribute to hepatocellular carcinoma (HCC) development, metastasis, and drug resistance. MSI2 and Notch1 signaling are involved in the maintenance of CSCs. However, it is unknown whether MSI2 and Notch1 are involved in the maintenance of CD44v6+ LCSCs. Therefore, we investigated the clinical significance and function of MSI2 and its relationship with Notch1 signaling in the maintenance of stemness properties in CD44v6+ LCSCs.

Methods

The expression of MSI2 and CD44v6 were detected by fresh specimens and a HCC tissue microarray. The tissue microarray containing 82 HCC samples was used to analyze the correlation between CD44v6 and MSI2. CD44v6+/− cells were isolated using microbeads sorting. We explored the roles of MSI2 and Notch1 signaling in CD44v6+ LCSCs by sphere formation assay, transwell assay, clone formation assay in vitro, and xenograft tumor models in vivo. A Notch RT² PCR Array, Co-immunoprecipitation, and RNA-immunoprecipitation were used to further investigate the molecular mechanism of MSI2 in activating Notch1 signaling.

Results

Here, we found MSI2 expression was positively correlated with high CD44v6 expression in HCC tissues, and further correlated with tumor differentiation. CD44v6+ cells isolated from HCC cell lines exhibited increased self-renewal, proliferation, migration and invasion, resistance to Sorafenib and tumorigenic capacity. Both MSI2 and Notch1 signaling were elevated in sorted CD44v6+ cells than CD44v6- cells and played essential roles in the maintenance of stemness of CD44v6+ LCSCs. Mechanically, MSI2 directly bound to Lunatic fringe (LFNG) mRNA and protein, resulting in Notch1 activation.

Conclusions

Our results demonstrated that MSI2 maintained the stemness of CD44v6+ LCSCs by activating Notch1 signaling through the interaction with LFNG, which could be a potential molecular target for stem cell-targeted therapy for liver cancer.

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Forner A, Reig M, Bruix J. Hepatocellular carcinoma. Lancet. 2018;391(10127):1301-14.CrossRef

Cheng Z, Li X, Ding J. Characteristics of liver cancer stem cells and clinical correlations. Cancer Lett. 2016;379(2):230–8.PubMedCrossRef

Sell S, Leffert HL. Liver cancer stem cells. J Clin Oncol. 2008;26(17):2800–5.PubMedCrossRef

Nio K, Yamashita T, Kaneko S. The evolving concept of liver cancer stem cells. Mol Cancer. 2017;16(1):4.PubMedPubMedCentralCrossRef

Visvader JE, Lindeman GJ. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 2008;8(10):755–68.PubMedCrossRef

Todaro M, Gaggianesi M, Catalano V, Benfante A, Iovino F, Biffoni M, Apuzzo T, Sperduti I, Volpe S, Cocorullo G, Gulotta G, Dieli F, De Maria R, Stassi G. CD44v6 is a marker of constitutive and reprogrammed cancer stem cells driving colon cancer metastasis. Cell Stem Cell. 2014;14(3):342–56.PubMedCrossRef

Ni J, Cozzi PJ, Hao JL, Beretov J, Chang L, Duan W, Shigdar S, Delprado WJ, Graham PH, Bucci J, Kearsley JH, Li Y. CD44 variant 6 is associated with prostate cancer metastasis and chemo−/radioresistance. Prostate. 2014;74(6):602–17.PubMedCrossRef

Ponta H, Sherman L, Herrlich PA. CD44: from adhesion molecules to signalling regulators. Nat Rev Mol Cell Biol. 2003;4(1):33–45.PubMedCrossRef

Zhu Z, Hao X, Yan M, Yao M, Ge C, Gu J, Li J. Cancer stem/progenitor cells are highly enriched in CD133+CD44+ population in hepatocellular carcinoma. Int J Cancer. 2010;126(9):2067–78.PubMed

10.

Okabe M, Imai T, Kurusu M, Hiromi Y, Okano H. Translational repression determines a neuronal potential in Drosophila asymmetric cell division. Nature. 2001;411(6833):94–8.PubMedCrossRef

11.

Ito T, Kwon HY, Zimdahl B, Congdon KL, Blum J, Lento WE, Zhao C, Lagoo A, Gerrard G, Foroni L, Goldman J, Goh H, Kim SH, Kim DW, Chuah C, Oehler VG, Radich JP, Jordan CT, Reya T. Regulation of myeloid leukaemia by the cell-fate determinant Musashi. Nature. 2010;466(7307):765–8.PubMedPubMedCentralCrossRef

12.

de Andres-Aguayo L, Varas F, Graf T. Musashi 2 in hematopoiesis. Curr Opin Hematol. 2012;19(4):268–72.PubMedCrossRef

13.

Nishimoto Y, Okano H. New insight into cancer therapeutics: induction of differentiation by regulating the Musashi/numb/notch pathway. Cell Res. 2010;20(10):1083–5.PubMedCrossRef

14.

Kudinov AE, Deneka A, Nikonova AS, Beck TN, Ahn YH, Liu X, Martinez CF, Schultz FA, Reynolds S, Yang DH, Cai KQ, Yaghmour KM, Baker KA, Egleston BL, Nicolas E, Chikwem A, Andrianov G, Singh S, Borghaei H, Serebriiskii IG, Gibbons DL, Kurie JM, Golemis EA, Boumber Y. Musashi-2 (MSI2) supports TGF-beta signaling and inhibits claudins to promote non-small cell lung cancer (NSCLC) metastasis. Proc Natl Acad Sci U S A. 2016;113(25):6955–60.PubMedPubMedCentralCrossRef

15.

Sheng W, Dong M, Chen C, Wang Z, Li Y, Wang K, Li Y, Zhou J. Cooperation of Musashi-2, numb, MDM2, and P53 in drug resistance and malignant biology of pancreatic cancer. FASEB J. 2017;31(6):2429–38.PubMedCrossRef

16.

Sheng W, Dong M, Chen C, Li Y, Liu Q, Dong Q. Musashi2 promotes the development and progression of pancreatic cancer by down-regulating numb protein. Oncotarget. 2017;8(9):14359–73.PubMedCrossRef

17.

He L, Zhou X, Qu C, Hu L, Tang Y, Zhang Q, Liang M, Hong J. Musashi2 predicts poor prognosis and invasion in hepatocellular carcinoma by driving epithelial-mesenchymal transition. J Cell Mol Med. 2014;18(1):49–58.PubMedCrossRef

18.

Wang MH, Qin SY, Zhang SG, Li GX, Yu ZH, Wang K, Wang B, Teng MJ, Peng ZH. Musashi-2 promotes hepatitis Bvirus related hepatocellular carcinoma progression via the Wnt/beta-catenin pathway. Am J Cancer Res. 2015;5(3):1089–100.PubMedPubMedCentral

19.

Lu J, Ye X, Fan F, Xia L, Bhattacharya R, Bellister S, Tozzi F, Sceusi E, Zhou Y, Tachibana I, Maru DM, Hawke DH, Rak J, Mani SA, Zweidler-McKay P, Ellis LM. Endothelial cells promote the colorectal cancer stem cell phenotype through a soluble form of Jagged-1. Cancer Cell. 2013;23(2):171–85.PubMedPubMedCentralCrossRef

20.

Androutsellis-Theotokis A, Leker RR, Soldner F, Hoeppner DJ, Ravin R, Poser SW, Rueger MA, Bae SK, Kittappa R, McKay RD. Notch signalling regulates stem cell numbers in vitro and in vivo. Nature. 2006;442(7104):823–6.PubMedCrossRef

21.

Fan X, Khaki L, Zhu TS, Soules ME, Talsma CE, Gul N, Koh C, Zhang J, Li YM, Maciaczyk J, Nikkhah G, Dimeco F, Piccirillo S, Vescovi AL, Eberhart CG. NOTCH pathway blockade depletes CD133-positive glioblastoma cells and inhibits growth of tumor neurospheres and xenografts. Stem cells (Dayton, Ohio). 2010;28(1):5–16.

22.

Man J, Yu X, Huang H, Zhou W, Xiang C, Huang H, Miele L, Liu Z, Bebek G, Bao S, Yu JS. Hypoxic Induction of Vasorin Regulates Notch1 Turnover to Maintain Glioma Stem-like Cells. Cell Stem Cell. 2018;22(1):104–18.e6.PubMedCrossRef

23.

Siebel C, Lendahl U. Notch signaling in development, tissue homeostasis, and disease. Physiol Rev. 2017;97(4):1235–94.PubMedCrossRef

24.

Harvey BM, Haltiwanger RS. Regulation of notch function by O-glycosylation. Adv Exp Med Biol. 2018;1066:59–78.PubMedCrossRef

25.

Wang R, Li Y, Tsung A, Huang H, Du Q, Yang M, Deng M, Xiong S, Wang X, Zhang L, Geller DA, Cheng B, Billiar TR. iNOS promotes CD24(+)CD133(+) liver cancer stem cell phenotype through a TACE/ADAM17-dependent notch signaling pathway. Proc Natl Acad Sci U S A. 2018;115(43):E10127–E36.PubMedPubMedCentralCrossRef

26.

Luo J, Wang P, Wang R, Wang J, Liu M, Xiong S, Li Y, Cheng B. The notch pathway promotes the cancer stem cell characteristics of CD90+ cells in hepatocellular carcinoma. Oncotarget. 2016;7(8):9525–37.PubMedCrossRef

27.

Wang R, Sun Q, Wang P, Liu M, Xiong S, Luo J, Huang H, Du Q, Geller DA, Cheng B. Notch and Wnt/beta-catenin signaling pathway play important roles in activating liver cancer stem cells. Oncotarget. 2016;7(5):5754–68.PubMed

28.

Fang T, Lv H, Wu F, Wang C, Li T, Lv G, Tang L, Guo L, Tang S, Cao D, Wu M, Yang W, Wang H. Musashi 2 contributes to the stemness and chemoresistance of liver cancer stem cells via LIN28A activation. Cancer Lett. 2017;384:50–9.PubMedCrossRef

29.

Fox RG, Park FD, Koechlein CS, Kritzik M, Reya T. Musashi signaling in stem cells and cancer. Annu Rev Cell Dev Biol. 2015;31:249–67.PubMedCrossRef

30.

Sakakibara S, Nakamura Y, Satoh H, Okano H. Rna-binding protein Musashi2: developmentally regulated expression in neural precursor cells and subpopulations of neurons in mammalian CNS. J Neurosci. 2001;21(20):8091–107.PubMedPubMedCentralCrossRef

31.

Kang MH, Jeong KJ, Kim WY, Lee HJ, Gong G, Suh N, Gyorffy B, Kim S, Jeong SY, Mills GB, Park YY. Musashi RNA-binding protein 2 regulates estrogen receptor 1 function in breast cancer. Oncogene. 2017;36(12):1745–52.PubMedCrossRef

32.

Choi SH, Kim AR, Nam JK, Kim JM, Kim JY, Seo HR, Lee HJ, Cho J, Lee YJ. Tumour-vasculature development via endothelial-to-mesenchymal transition after radiotherapy controls CD44v6(+) cancer cell and macrophage polarization. Nat Commun. 2018;9(1):5108.PubMedPubMedCentralCrossRef

33.

Endo K, Terada T. Protein expression of CD44 (standard and variant isoforms) in hepatocellular carcinoma: relationships with tumor grade, clinicopathologic parameters, p53 expression, and patient survival. J Hepatol. 2000;32(1):78–84.PubMedCrossRef

34.

Zhou ZJ, Dai Z, Zhou SL, Fu XT, Zhao YM, Shi YH, Zhou J, Fan J. Overexpression of HnRNP A1 promotes tumor invasion through regulating CD44v6 and indicates poor prognosis for hepatocellular carcinoma. Int J Cancer. 2013;132(5):1080–9.PubMedCrossRef

35.

Ma L, Dong L, Chang P. CD44v6 engages in colorectal cancer progression. Cell Death Dis. 2019;10(1):30.PubMedPubMedCentralCrossRef

36.

Park SM, Gonen M, Vu L, Minuesa G, Tivnan P, Barlowe TS, Taggart J, Lu Y, Deering RP, Hacohen N, Figueroa ME, Paietta E, Fernandez HF, Tallman MS, Melnick A, Levine R, Leslie C, Lengner CJ, Kharas MG. Musashi2 sustains the mixed-lineage leukemia-driven stem cell regulatory program. J Clin Invest. 2015;125(3):1286–98.PubMedPubMedCentralCrossRef

37.

Taggart J, Ho TC, Amin E, Xu H, Barlowe TS, Perez AR, Durham BH, Tivnan P, Okabe R, Chow A, Vu L, Park SM, Prieto C, Famulare C, Patel M, Lengner CJ, Verma A, Roboz G, Guzman M, Klimek VM, Abdel-Wahab O, Leslie C, Nimer SD, Kharas MG. MSI2 is required for maintaining activated myelodysplastic syndrome stem cells. Nat Commun. 2016;7:10739.PubMedPubMedCentralCrossRef

38.

Kharas MG, Lengner CJ, Al-Shahrour F, Bullinger L, Ball B, Zaidi S, Morgan K, Tam W, Paktinat M, Okabe R, Gozo M, Einhorn W, Lane SW, Scholl C, Frohling S, Fleming M, Ebert BL, Gilliland DG, Jaenisch R, Daley GQ. Musashi-2 regulates normal hematopoiesis and promotes aggressive myeloid leukemia. Nat Med. 2010;16(8):903–8.PubMedPubMedCentralCrossRef

39.

Wang J, Sullenger BA, Rich JN. Notch signaling in cancer stem cells. Adv Exp Med Biol. 2012;727:174–85.PubMedCrossRef

40.

Fang S, Liu M, Li L, Zhang FF, Li Y, Yan Q, Cui YZ, Zhu YH, Yuan YF, Guan XY. Lymphoid enhancer-binding factor-1 promotes stemness and poor differentiation of hepatocellular carcinoma by directly activating the NOTCH pathway. Oncogene. 2019;38(21):4061–74.PubMedCrossRef

41.

Griner LN, Reuther GW. Aggressive myeloid leukemia formation is directed by the Musashi 2/numb pathway. Cancer Biol Ther. 2010;10(10):979–82.PubMedCrossRef

42.

Zhang S, Chung WC, Xu K. Lunatic fringe is a potent tumor suppressor in Kras-initiated pancreatic cancer. Oncogene. 2016;35(19):2485–95.PubMedCrossRef

43.

Zhang S, Chung WC, Wu G, Egan SE, Xu K. Tumor-suppressive activity of Lunatic Fringe in prostate through differential modulation of Notch receptor activation. Neoplasia (New York, NY). 2014;16(2):158–67.CrossRef

44.

Chung WC, Zhang S, Challagundla L, Zhou Y, Xu K. Lunatic Fringe and p53 Cooperatively Suppress Mesenchymal Stem-Like Breast Cancer. Neoplasia (New York, NY). 2017;19(11):885–95.CrossRef

45.

Tsukumo S, Hirose K, Maekawa Y, Kishihara K, Yasutomo K. Lunatic fringe controls T cell differentiation through modulating notch signaling. J Immunol. 2006;177(12):8365–71.PubMedCrossRef

46.

Kato TM, Kawaguchi A, Kosodo Y, Niwa H, Matsuzaki F. Lunatic fringe potentiates notch signaling in the developing brain. Mol Cell Neurosci. 2010;45(1):12–25.PubMedCrossRef

Title: Musashi2 contributes to the maintenance of CD44v6+ liver cancer stem cells via notch1 signaling pathway
Authors: Xiju Wang
Ronghua Wang
Shuya Bai
Si Xiong
Yawen Li
Man Liu
Zhenxiong Zhao
Yun Wang
Yuchong Zhao
Wei Chen
Timothy R. Billiar
Bin Cheng
Publication date: 01-12-2019
Publisher: BioMed Central
Keywords: Hepatocellular Carcinoma
Hepatocellular Carcinoma
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2019
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/s13046-019-1508-1

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