Top

Published in:

Open Access 01-12-2016 | Research article

Fusion with stem cell makes the hepatocellular carcinoma cells similar to liver tumor-initiating cells

Authors: Ran Wang, Shuxun Chen, Changxian Li, Kevin Tak Pan Ng, Chi-wing Kong, Jinping Cheng, Shuk Han Cheng, Ronald A. Li, Chung Mau Lo, Kwan Man, Dong Sun

Published in: BMC Cancer | Issue 1/2016

Abstract

Background

Cell fusion is a fast and highly efficient technique for cells to acquire new properties. The fusion of somatic cells with stem cells can reprogram somatic cells to a pluripotent state. Our research on the fusion of stem cells and cancer cells demonstrates that the fused cells can exhibit stemness and cancer cell-like characteristics. Thus, tumor-initiating cell-like cells are generated.

Methods

We employed laser-induced single-cell fusion technique to fuse the hepatocellular carcinoma cells and human embryonic stem cells (hESC). Real-time RT-PCR, flow cytometry and in vivo tumorigenicity assay were adopted to identify the gene expression difference.

Results

We successfully produced a fused cell line that coalesces the gene expression information of hepatocellular carcinoma cells and stem cells. Experimental results showed that the fused cells expressed cancer and stemness markers as well as exhibited increased resistance to drug treatment and enhanced tumorigenesis.

Conclusions

Fusion with stem cells transforms liver cancer cells into tumor initiating-like cells. Results indicate that fusion between cancer cell and stem cell may generate tumor initiating-like cells.

Ficz G, Reik W. Reprogramming by cell fusion: boosted by Tets. Mol Cell. 2013;49(6):1017–8. doi:10.1016/j.molcel.2013.03.014.CrossRefPubMed

Ferrand J, Noel D, Lehours P, Prochazkova-Carlotti M, Chambonnier L, Menard A, et al. Human bone marrow-derived stem cells acquire epithelial characteristics through fusion with gastrointestinal epithelial cells. PLoS One. 2011;6(5):e19569. doi:10.1371/journal.pone.0019569.PubMedCentralCrossRefPubMed

Piccolo FM, Bagci H, Brown KE, Landeira D, Soza-Ried J, Feytout A, et al. Different roles for Tet1 and Tet2 proteins in reprogramming-mediated erasure of imprints induced by EGC fusion. Mol Cell. 2013;49(6):1023–33. doi:10.1016/j.molcel.2013.01.032.PubMedCentralCrossRefPubMed

Yamanaka S, Blau HM. Nuclear reprogramming to a pluripotent state by three approaches. Nature. 2010;465(7299):704–12. doi:10.1038/Nature09229.PubMedCentralCrossRefPubMed

Forbes SJ, Newsome PN. New horizons for stem cell therapy in liver disease. J Hepatol. 2012;56(2):496–9. doi:10.1016/j.jhep.2011.06.022.CrossRefPubMed

Sakaida I, Terai S, Yamamoto N, Aoyama K, Ishikawa T, Nishina H, et al. Transplantation of bone marrow cells reduces CCl4-induced liver fibrosis in mice. Hepatology. 2004;40(6):1304–11. doi:10.1002/hep.20452.CrossRefPubMed

Furst G, Schulte am Esch J, Poll LW, Hosch SB, Fritz LB, Klein M, et al. Portal vein embolization and autologous CD133+ bone marrow stem cells for liver regeneration: initial experience. Radiology. 2007;243(1):171–9. doi:10.1148/radiol.2431060625.CrossRefPubMed

Newsome PN, Johannessen I, Boyle S, Dalakas E, McAulay KA, Samuel K, et al. Human cord blood-derived cells can differentiate into hepatocytes in the mouse liver with no evidence of cellular fusion. Gastroenterology. 2003;124(7):1891–900. doi:10.1016/S0016-5085(03)00401-3.CrossRefPubMed

Visvader JE, Lindeman GJ. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 2008;8(10):755–68. doi:10.1038/nrc2499.CrossRefPubMed

10.

Dang H, Ding W, Emerson D, Rountree CB. Snail1 induces epithelial-to-mesenchymal transition and tumor initiating stem cell characteristics. BMC Cancer. 2011;11:396. doi:10.1186/1471-2407-11-396.PubMedCentralCrossRefPubMed

11.

Yamashita T, Wang XW. Cancer stem cells in the development of liver cancer. J Clin Invest. 2013;123(5):1911–8. doi:10.1172/JCI66024.PubMedCentralCrossRefPubMed

12.

Yamashita T, Kaneko S. Orchestration of hepatocellular carcinoma development by diverse liver cancer stem cells. J Gastroenterol. 2014;49(7):1105–10. doi:10.1007/s00535-014-0951-1.CrossRefPubMed

13.

Zhang F, Chen XP, Zhang W, Dong HH, Xiang S, Zhang WG, et al. Combined hepatocellular cholangiocarcinoma originating from hepatic progenitor cells: immunohistochemical and double-fluorescence immunostaining evidence. Histopathology. 2008;52(2):224–32. doi:10.1111/j.1365-2559.2007.02929.x.CrossRefPubMed

14.

Chen SX, Cheng JP, Kong CW, Wang XL, Cheng SH, Li RA, et al. Laser-induced fusion of human embryonic stem cells with optical tweezers. Appl Phys Lett. 2013;103(3):033701. doi:10.1063/1.4813822.CrossRef

15.

Wagner W, Ansorge A, Wirkner U, Eckstein V, Schwager C, Blake J, et al. Molecular evidence for stem cell function of the slow-dividing fraction among human hematopoietic progenitor cells by genome-wide analysis. Blood. 2004;104(3):675–86. doi:10.1182/blood-2003-10-3423.CrossRefPubMed

16.

Mima K, Okabe H, Ishimoto T, Hayashi H, Nakagawa S, Kuroki H, et al. CD44s regulates the TGF-beta-mediated mesenchymal phenotype and is associated with poor prognosis in patients with hepatocellular carcinoma. Cancer Res. 2012;72(13):3414–23. doi:10.1158/0008-5472.CAN-12-0299.CrossRefPubMed

17.

Wang XW, Seed B. A PCR primer bank for quantitative gene expression analysis. Nucleic Acids Res. 2003;31(24):e154. doi:10.1093/nar/gng154.PubMedCentralCrossRefPubMed

18.

Yamashita T, Wang XW. Cancer stem cells in the development of liver cancer. J Clin Invest. 2013;123(5):1911–8. doi:10.1172/JCI66024.PubMedCentralCrossRefPubMed

19.

Yao J, Zhang T, Ren J, Yu M, Wu G. Effect of CD133/prominin-1 antisense oligodeoxynucleotide on in vitro growth characteristics of Huh-7 human hepatocarcinoma cells and U251 human glioma cells. Oncol Rep. 2009;22(4):781–7.PubMed

20.

Patrawala L, Calhoun-Davis T, Schneider-Broussard R, Tang DG. Hierarchical organization of prostate cancer cells in xenograft tumors: The CD44(+)alpha 2 beta 1(+) cell population is enriched in tumor-initiating cells. Cancer Res. 2007;67(14):6796–805. doi:10.1158/0008-5472.Can-07-0490.CrossRefPubMed

21.

Yamashita T, Ji J, Budhu A, Forgues M, Yang W, Wang HY, et al. EpCAM-positive hepatocellular carcinoma cells are tumor-initiating cells with stem/progenitor cell features. Gastroenterology. 2009;136(3):1012–24. doi:10.1053/j.gastro.2008.12.004.PubMedCentralCrossRefPubMed

22.

Ji JF, Yamashita T, Budhu A, Forgues M, Jia HL, Li CL, et al. Identification of MicroRNA-181 by genome-wide screening as a critical player in EpCAM-positive hepatic cancer stem cells. Hepatology. 2009;50(2):472–80. doi:10.1002/Hep.22989.PubMedCentralCrossRefPubMed

23.

Gostner JM, Fong D, Wrulich OA, Lehne F, Zitt M, Hermann M, et al. Effects of EpCAM overexpression on human breast cancer cell lines. BMC Cancer. 2011;11:45. doi:10.1186/1471-2407-11-45.PubMedCentralCrossRefPubMed

24.

Dean M, Fojo T, Bates S. Tumour stem cells and drug resistance. Nat Rev Cancer. 2005;5(4):275–84. doi:10.1038/nrc1590.CrossRefPubMed

25.

Houthuijzen JM, Daenen LG, Roodhart JM, Voest EE. The role of mesenchymal stem cells in anti-cancer drug resistance and tumour progression. Br J Cancer. 2012;106(12):1901–6. doi:10.1038/bjc.2012.201.PubMedCentralCrossRefPubMed

26.

Abdullah LN, Chow EK. Mechanisms of chemoresistance in cancer stem cells. Clin Translat Med. 2013;2(1):3. doi:10.1186/2001-1326-2-3.CrossRef

27.

Enyedy IJ, Ling Y, Nacro K, Tomita Y, Wu XH, Cao YY, et al. Discovery of small-molecule inhibitors of bcl-2 through structure-based computer screening. J Med Chem. 2001;44(25):4313–24. doi:10.1021/Jm010016f.CrossRefPubMed

28.

van Oosterwijk JG, Herpers B, Meijer D, Briaire-de Bruijn IH, Cleton-Jansen AM, Gelderblom H, et al. Restoration of chemosensitivity for doxorubicin and cisplatin in chondrosarcoma in vitro: BCL-2 family members cause chemoresistance. Ann Oncol. 2012;23(6):1617. doi:10.1093/annonc/mdr512.CrossRefPubMed

29.

Siddique HR, Parray A, Tarapore RS, Wang L, Mukhtar H, Karnes RJ, et al. BMI1 polycomb group protein acts as a master switch for growth and death of tumor cells: regulates TCF4-transcriptional factor-induced BCL2 signaling. PLoS One. 2013;8(5):e60664. doi:10.1371/journal.pone.0060664.PubMedCentralCrossRefPubMed

30.

Ji J, Wang XW. Clinical implications of cancer stem cell biology in hepatocellular carcinoma. Semin Oncol. 2012;39(4):461–72. doi:10.1053/j.seminoncol.2012.05.011.PubMedCentralCrossRefPubMed

31.

Kobayashi I, Matsuo K, Ishibashi Y, Kanda S, Sakai H. The proliferative activity in dysplasia and carcinoma in situ of the uterine cervix analyzed by proliferating cell nuclear antigen immunostaining and silver-binding argyrophilic nucleolar organizer region staining. Hum Pathol. 1994;25(2):198–202.CrossRefPubMed

32.

Inwald EC, Klinkhammer-Schalke M, Hofstadter F, Zeman F, Koller M, Gerstenhauer M, et al. Ki-67 is a prognostic parameter in breast cancer patients: results of a large population-based cohort of a cancer registry. Breast Cancer Res Tr. 2013;139(2):539–52. doi:10.1007/s10549-013-2560-8.CrossRef

33.

Luporsi E, Andre F, Spyratos F, Martin PM, Jacquemier J, Penault-Llorca F, et al. Ki-67: level of evidence and methodological considerations for its role in the clinical management of breast cancer: analytical and critical review. Breast Cancer Res Tr. 2012;132(3):895–915. doi:10.1007/s10549-011-1837-z.CrossRef

34.

Lluis F, Cosma MP. Cell-fusion-mediated somatic-cell reprogramming: a mechanism for tissue regeneration. J Cell Physiol. 2010;223(1):6–13. doi:10.1002/Jcp.22003.PubMed

35.

Silva J, Chambers I, Pollard S, Smith A. Nanog promotes transfer of pluripotency after cell fusion. Nature. 2006;441(7096):997–1001. doi:10.1038/nature04914.CrossRefPubMed

36.

Wong CC, Gaspar-Maia A, Ramalho-Santos M, Reijo Pera RA. High-efficiency stem cell fusion-mediated assay reveals Sall4 as an enhancer of reprogramming. Plos One. 2008;3(4):e1955. doi:10.1371/journal.pone.0001955.PubMedCentralCrossRefPubMed

37.

Cusulin C, Monni E, Ahlenius H, Wood J, Brune JC, Lindvall O, et al. Embryonic stem cell-derived neural stem cells fuse with microglia and mature neurons. Stem Cells. 2012;30(12):2657–71. doi:10.1002/Stem.1227.CrossRefPubMed

Title: Fusion with stem cell makes the hepatocellular carcinoma cells similar to liver tumor-initiating cells
Authors: Ran Wang
Shuxun Chen
Changxian Li
Kevin Tak Pan Ng
Chi-wing Kong
Jinping Cheng
Shuk Han Cheng
Ronald A. Li
Chung Mau Lo
Kwan Man
Dong Sun
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: BMC Cancer / Issue 1/2016
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-016-2094-7

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2016

Sentinel lymph node biopsy after neoadjuvant chemotherapy for breast cancer: retrospective comparative evaluation of clinically axillary lymph node positive and negative patients, including those with axillary lymph node metastases confirmed by fine needle aspiration

The validity of pediatric cancer diagnoses in a population-based general cancer registry in Ontario, Canada

Interplay of N-Cadherin and matrix metalloproteinase 9 enhances human nasopharyngeal carcinoma cell invasion

Nanoroughened adhesion-based capture of circulating tumor cells with heterogeneous expression and metastatic characteristics

Chemotherapy and radiation therapy elicits tumor specific T cell responses in a breast cancer patient

Pleural fluid osteopontin, vascular endothelial growth factor, and urokinase-type plasminogen activator levels as predictors of pleurodesis outcome and prognosticators in patients with malignant pleural effusion: a prospective cohort study

Keynote webinar | Spotlight on antibody–drug conjugates in cancer