CD133+CXCR4+ colon cancer cells exhibit metastatic potential and predict poor prognosis of patients

BMC Medicine

Li-xin Wei²  Li Zhang³  Hou-jia Liu³  Yao-cheng Rui¹  En-da Yu³  Xiao-dong Xu³  Xue Zhao²  Yang Yang²  Rong Li²  Yang Wang³  Hao Wang³  Tie-jun Li¹  Shuang-fen Tao²  Ying-ying Jing²  Zhi-peng Han²  Shan-shan Zhang² 

[1] Department of Pharmacology, Second Military Medical University, 325 Guohe Road, Shanghai 200433, China;Tumor Immunology and Gene Therapy Center, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, 225 Changhai Road, Shanghai 200438, China;Changhai Hospital, Second Military Medical University, 168 Changhai Road, Shanghai 200433, China

关键词: liver metastasis;    epithelial-mesenchymal transition;    CXCR4;    cancer stem cell;    colorectal cancer;   

Others  :  1125903
DOI  :  10.1186/1741-7015-10-85

 received in 2011-10-31, accepted in 2012-08-07,  发布年份 2012

【 摘 要 】

Background

Colorectal cancer (CRC), which frequently metastasizes to the liver, is one of the three leading causes of cancer-related deaths worldwide. Growing evidence suggests that a subset of cells exists among cancer stem cells. This distinct subpopulation is thought to contribute to liver metastasis; however, it has not been fully explored in CRC yet.

Methods

Flow cytometry analysis was performed to detect distinct subsets with CD133 and CXCR4 markers in human primary and metastatic CRC tissues. The 'stemness' and metastatic capacities of different subpopulations derived from the colon cancer cell line HCT116 were compared in vitro and in vivo. The roles of epithelial-mesenchymal transition (EMT) and stromal-cell derived factor-1 (SDF-1) in the metastatic process were also investigated. A survival curve was used to explore the correlation between the content of CD133⁺CXCR4⁺ cancer cells and patient survival.

Results

In human specimens, the content of CD133⁺CXCR4⁺ cells was higher in liver metastases than in primary colorectal tumors. Clonogenic and tumorigenic cells were restricted to CD133⁺ cells in the HCT116 cell line, with CXCR4 expression having no impact on the 'stemness' properties. We found that CD133⁺CXCR4⁺ cancer cells had a high metastatic capacity in vitro and in vivo. Compared with CD133⁺CXCR4^- cells, CD133⁺CXCR4⁺ cancer cells experienced EMT, which contributed partly to their metastatic phenotype. We then determined that SDF-1/CXCL12 treatment could further induce EMT in CD133⁺CXCR4⁺ cancer cells and enhance their invasive behavior, while this could not be observed in CD133⁺CXCR4^- cancer cells. Blocking SDF-1/CXCR4 interaction with a CXCR4 antagonist, AMD3100 (1,10-[1,4-phenylenebis(methylene)]bis-1,4,8,11 -tetraazacyclotetradecane octahydrochloride), inhibited metastatic tumor growth in a mouse hepatic metastasis model. Finally, a high percentage of CD133⁺CXCR4⁺ cells in human primary CRC was associated with a reduced two-year survival rate.

Conclusions

Strategies targeting the SDF-1/CXCR4 interaction may have important clinical applications in the suppression of colon cancer metastasis. Further investigations on how high expression of CXCR4 and EMT occur in this identified cancer stem cell subset are warranted to provide insights into our understanding of tumor biology.

【 授权许可】

   
2012 Zhang et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Ries LAG, Eisner MP, Kosary CL: SEER Cancer Statistics Review. Bethesda, MD: National Cancer Institute; 2002.
• [2]Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ: Cancer statistics. CA Cancer J Clin 2008, 58:71-96.
• [3]Clarke MF, Dick JE, Dirks PB, Eaves CJ, Jamieson CHM, Jones DL, Visvader J, Weissman IL, Wahl GM: Cancer stem cells--perspectives on current status and future directions: AACR Workshop on Cancer Stem Cells. Cancer Res 2006, 66:9339-9344.
• [4]Tannishtha R, Sean JM, Michael FC, Irving LW: Stem cells, cancer, and cancer stem cells. Nature 2001, 414:105-111.
• [5]Tang C, Ang BT, Pervaiz S: Cancer stem cell: target for anti-cancer therapy. FASEB J 2007, 21:3777-85.
• [6]Bonnet D, Dick JE: Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med 1997, 3:730-737.
• [7]Al-Hajj M, Wicha MS, Benito-Hernandez A: Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA 2003, 100:3983-3988.
• [8]Collins AT, Berry PA, Hyde C: Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res 2005, 65:10946-10951.
• [9]Prince ME, Sivanandan R, Kaczorowski A: Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci USA 2007, 104:973-978.
• [10]Singh SK, Hawkins C, Clarke ID: Identification of human brain tumour initiating cells. Nature 2004, 432:396-401.
• [11]O'Brien CA, Pollett A, Gallinger S: A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 2007, 445:106-110.
• [12]Ricci-Vitiani L, Lombardi DG, Pilozzi E: Identification and expansion of human colon cancer-initiating cells. Nature 2007, 445:111-115.
• [13]Brabletz T, Jung A, Spaderna S: Opinion: migrating cancer stem cells -- an integrated concept of malignant tumour progression. Nat Rev Cancer 2005, (5:):744-749.
• [14]Stetler-Stevenson WG, Kleiner DE Jr: Molecular biology of cancer: invasion and metastasis. In Cancer: Principles and Practice of Oncology. Edited by DeVita VT Jr, Hellman S, Rosenberg SA. Philadelphia, PA: Lippincott Williams &Wilkins; 2001:123-136.
• [15]Kucia M, Reca R, Miekus K: Trafficking of normal stem cells and metastasis of cancer stem cells involve similar mechanisms: pivotal role of the SDF-1-CXCR4 axis. Stem Cells 2005, 23:879-894.
• [16]Muller A, Homey B, Soto H: Involvement of chemokine receptors in breast cancer metastasis. Nature 2001, 410:50-56.
• [17]Saur D, Seidler B, Schneider G: CXCR4 expression increases liver and lung metastasis in a mouse model of pancreatic cancer. Gastroenterology 2005, 129:1237-1250.
• [18]Zeelenberg IS, Ruuls-Van Stalle L, Roos E: The chemokine receptor CXCR4 is required for outgrowth of colon carcinoma micrometastases. Cancer Res 2003, 63:3833-3839.
• [19]Kim J, Takeuchi H, Lam ST: Chemokine receptor CXCR4 expression in colorectal cancer patients increases the risk for recurrence and for poor survival. J Clin Oncol 2005, 23:2744-2753.
• [20]Schimanski CC, Schwald S, Simiantonaki N: Effect of chemokine receptors CXCR4 and CCR7 on the metastatic behavior of human colorectal cancer. Clin Cancer Res 2005, 11:1743-1750.
• [21]Hermann PC, Huber SL, Herrler T: Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 2007, 1:313-323.
• [22]Gavert N, Ben-Ze'ev A: Epithelial-mesenchymal transition and the invasive potential of tumors. Trends Mol Med 2008, 14:199-209.
• [23]Thiery JP, Sleeman JP: Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol 2006, 7:131-142.
• [24]Gilles C, Thompson EW: The epithelial to mesenchymal transition and metastatic progression in carcinoma. Breast J 1996, 2:83-96.
• [25]Thiery JP: Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer 2002, 2:442-454.
• [26]Onoue T, Uchida D, Begum NM: Epithelial-mesenchymal transition induced by the stromal cell-derived factor-1/CXCR4 system in oral squamous cell carcinoma cells. Int J Oncol 2006, 29:1133-1138.
• [27]Na DC, Lee JE, Yoo JE: Invasion and EMT-associated genes are up-regulated in B viral hepatocellular carcinoma with high expression of CD133-human and cell culture study. Exp Mol Pathol 2011, 90:66-73.
• [28]Mani SA, Guo W, Liao MJ: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 2008, 133:704-715.
• [29]Morel AP, Lievre M, Thomas C: Generation of breast cancer stem cells through epithelial-mesenchymal transition. PLoS One 2008, 3:e2888.
• [30]Lo JF, Yu CC, Chiou SH: The epithelial-mesenchymal transition mediator S100A4 maintains cancer initiating cells in head and neck cancers. Cancer Res 2011, 71:1912-1923.
• [31]Li G, Liu C, Yuan J: CD133+ single cell-derived progenies of colorectal cancer cell line SW480 with different invasive and metastatic potential. Clin Exp Metastasis 2010, 27:517-527.
• [32]Hiraoka K, Kimura T, Logg CR: Therapeutic efficacy of replication-competent retrovirus vector-mediated suicide gene therapy in a multifocal colorectal cancer metastasis model. Cancer Res 2007, 67:5345-5353.
• [33]Ludbrook J, Dudley H: Why permutation tests are superior to t and F tests in biomedical research. Am Stat 1998, 52:127-132.
• [34]Hesterberg T, Moore DS, Monaghan S, Clipson A, Epstein R: Bootstrap methods and permutation tests. In Introduction to the Practice of Statistics. Volume 14. 5th edition. Edited by Moore DS, MacCabe GP. New York: WH Freeman & Company; 2005::1-70.
• [35]Marozzi M: A bi-aspect nonparametric test for the two-sample location problem. Comput Stat Data Anal 2004, 44:639-648.
• [36]Dunn OJ, Clark VA: Basic Statistics: A Primer for the Biomedical Sciences. 4th edition. (Wiley Series in Probability and Statistics). Hoboken, NJ: John Wiley & Sons; 2009.
• [37]He J, Jin ZC, Yu DH: Statistical reporting in Chinese biomedical journals. Lancet 2009, 373:2091-2093.
• [38]Thiery JP: Epithelial-mesenchymal transition in tumour progression. Nat Rev Cancer 2002, 2:442-454.
• [39]LaBarge MA, Bissell MJ: Is CD133 a marker of metastatic colon cancer stem cells? J Clin Invest 2008, 118:2021-2024.
• [40]Zhi-Li Y, Qi Z, Jun Y: Upregulated CD133 expression in tumorigenesis of colon cancer cells. World J Gastroenterol 2011, 17:932-937.
• [41]Pang R, Law WL, Chu AC: A subpopulation of CD26+ cancer stem cells with metastatic capacity in human colorectal cancer. Cell Stem Cell 2010, 6:603-615.
• [42]Ruoslahti E: How cancer spreads. Sci Am 1996, 275:72-77.
• [43]Elkin M, Vlodavsky I: Tail vein assay of cancer metastasis. Curr Protoc Cell Biol 2001, Chapter 19:Unit 19.2.
• [44]Johnson JL, Pillai S, Pernazza D, Sebti SM, Lawrence NJ, Chellappan SP: Regulation of matrix metalloproteinase genes by E2F transcription factors: Rb-Raf-1 interaction as a novel target for metastatic disease. Cancer Res 2012, 72:516-526.
• [45]Yang CH, Yue J, Pfeffer SR, Handorf CR, Pfeffer LM: MicroRNA miR-21 regulates the metastatic behavior of B16 melanoma cells. J Biol Chem 2011, 286:39172-39178.
• [46]Batlle E, Sancho E, Franci C: The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol 2000, 2:84-89.
• [47]Gavert N, Ben-Ze'ev A: Epithelial-mesenchymal transition and the invasive potential of tumors. Trends Mol Med 2008, 14:199-209.
• [48]Bertran E, Caja L, Navarro E: Role of CXCR4/SDF-1α in the migratory phenotype of hepatoma cells that have undergone epithelial-mesenchymal transition in response to the transforming growth factor-β. Cell Signal 2009, 21:1595-1606.
• [49]Taki M, Higashikawa K, Yoneda S: Up-regulation of stromal cell-derived factor-1α and its receptor CXCR4 expression accompanied with epithelial-mesenchymal transition in human oral squamous cell carcinoma. Oncol Rep 2008, 19:993-998.
• [50]Hirschmann-Jax C, Foster AE, Wulf GG: A distinct "side population" of cells with high drug efflux capacity in human tumor cells. Proc Natl Acad Sci USA 2004, 101:14228-14233.
• [51]Zhou S, Schuetz JD, Bunting KD: The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med 2001, 7:1028-1034.
• [52]Liu G, Yuan X, Zeng Z: Analysis of gene expression and chemoresistance of CD133+ cancer stem cells in glioblastoma. Mol Cancer 2006, 5:67. BioMed Central Full Text
• [53]Muller A, Homey B, Soto H, Ge N, Catron D, Buchanan ME, McClanahan T, Murphy E, Yuan W, Wagner SN, Barrera JL, Mohar A, Verastegui E, Zlotnik A: Involvement of chemokine receptors in breast cancer metastasis. Nature 2001, 410:50-56.
• [54]Saur D, Seidler B, Schneider G: CXCR4 expression increases liver and lung metastasis in a mouse model of pancreatic cancer. Gastroenterology 2005, 129:1237-1250.