Identification of CD44+CD24+ gastric cancer stem cells

  • Chaojun Zhang
  • Chenwei Li
  • Fengtian He
  • Yujiao Cai
  • Hua Yang
Original Paper

Abstract

Objective

Purification and characterization of cancer stem cells (CSCs) can lead to the identification of targets for therapeutic interventions of cancer. With regard to gastric cancer, studies have not yet defined and characterized CSCs.

Methods

The expression of the cell surface markers CD44 and CD24 was examined in gastric cell lines AGS and gastric cancer tissues from five patients with fluorescence-activated cell sorting analysis (FACS). The tumorigenic properties, self-renewal, and differentiated progeny in the two distinct cell populations CD44+CD24+ and CD44−CD24− were identified in vivo serial transplantation and in vitro culture. Real-time RT-PCR was used to assess the expression of sonic hedgehog (SHH), patched 1 (PTCH1), and GLI3 signaling molecules in CD44+CD24+ and CD44−CD24− cells.

Results

As few as 200 CD44+CD24+ cells injected in NOD–SCID mice were able to generate tumors in 50% of mice (6 of 12), while tumors did not form in mice until at least 10,000 CD44−CD24− cells were injected, where only one of 12 mice formed a tumor, further verifying that CD44+CD24+ gastric cancer cells have the capacity to both self-renew and produce differentiated progeny. Moreover, SHH, PTCH1, and GLI3 mRNA expression increased significantly in the CD44+CD24+ subpopulation when compared with the CD44−CD24− subpopulation.

Conclusions

These studies strongly suggest that the CD44+CD24+ subpopulation of human gastric cancer cell lines, AGS, is gastric cancer stem cells.

Keywords

Cancer stem cells Gastric cancer Cell surface marker Hedgehog signal 

References

  1. Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA 100:3983–3988PubMedCrossRefGoogle Scholar
  2. Bonnet D, Dick JE (1997) Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med 3:730–737PubMedCrossRefGoogle Scholar
  3. Charafe-Jauffret E, Monville F, Ginestier C, Dontu G, Birnbaum D, Wicha MS (2008) Cancer stem cells in breast: current opinion and future challenges. Pathobiology 75:75–84PubMedCrossRefGoogle Scholar
  4. Collins AT, Berry PA, Hyde C, Stower MJ, Maitland NJ (2005) Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res 65:10946–10951Google Scholar
  5. Dontu G, Jackson KW, McNicholas E, Kawamura MJ, Abdallah WM, Wicha MS (2004) Role of Notch signaling in cell-fate determination of human mammary stem/progenitor cells. Breast Cancer Res 6:R605–R615PubMedCrossRefGoogle Scholar
  6. Fang D, Nguyen TK, Leishear K, Finko R, Kulp AN, Hotz S et al (2005) A tumorigenic subpopulation with stem cell properties in melanomas. Cancer Res 65:9328–9337PubMedCrossRefGoogle Scholar
  7. Haraguchi N, Utsunomiya T, Inoue H, Tanaka F, Mimori K, Barnard GF, Mori M (2006) Characterization of a side population of cancer cells from human gastrointestinal system. Stem Cells 24:506–513PubMedCrossRefGoogle Scholar
  8. Hatsell S, Frost AR (2007) Hedgehog signaling in mammary gland development and breast cancer. J Mammary Gland Biol Neoplasia 129:163–173CrossRefGoogle Scholar
  9. Hsu KH, Tsai HW, Shan YS, Lin PW (2007) Significance of CD44 expression in gastrointestinal stromal tumors in relation to disease progression and survival. World J Surg 31:1438–1444PubMedCrossRefGoogle Scholar
  10. Karhadkar SS, Bova GS, Abdallah N, Dhara S et al (2004) Hedgehog signalling in prostate regeneration, neoplasia and metastasis. Nature 431:707–712PubMedCrossRefGoogle Scholar
  11. Katoh Y, Katoh M (2005) Hedgehog signaling pathway and gastric cancer. Cancer Biol Therapy 4:1050–1054CrossRefGoogle Scholar
  12. Lee SY, Han HS, Lee KY, Hwang TS et al (2007) Sonic hedgehog expression in gastric cancer and gastric adenoma. Oncol Rep 17:1051–1055PubMedGoogle Scholar
  13. Li C, Heidt DG, Dalerba P, Burant CF, Zhang L, Adsay V, Wicha M, Clarke MF, Simeone DM (2007) Identification of pancreatic cancer stem cells. Cancer Res 67:1030–1037PubMedCrossRefGoogle Scholar
  14. Liu S, Dontu G, Mantle ID, Patel S, Ahn NS, Jackson KW, Suri P, Wicha MS (2006) Hedgehog signaling and Bmi-1 regulate self-renewal of normal and malignant human mammary stem cells. Cancer Res 66:6063–6071PubMedCrossRefGoogle Scholar
  15. Ma X, Chen K, Huang S, Zhang X, Adegboyega PA et al (2005) Frequent activation of the hedgehog pathway in advanced gastric adenocarcinomas. Carcinogenesis 26:1698–1705PubMedCrossRefGoogle Scholar
  16. Matsui W, Huff CA, Wang Q, Malehorn MT, Barber J, Tanhehco Y, Smith BD, Civin CI, Jones RJ (2004) Characterization of clonogenic multiple myeloma cells. Blood 103:2332–2336PubMedCrossRefGoogle Scholar
  17. O’Brien CA, Pollett A, Gallinger S, Dick JE (2007) A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445:106–110Google Scholar
  18. Parkin DM, Bray F, Ferlay J, Pisani P (2005) Global cancer statistics, 2002. CA Cancer J Clin 55:74–108PubMedCrossRefGoogle Scholar
  19. Peacock CD, Wang Q, Gesell GS et al (2007) Hedgehog signaling maintains a tumor stem cell compartment in multiple myeloma. Proc Natl Acad Sci USA 104:4048–4053PubMedCrossRefGoogle Scholar
  20. Phillips TM, McBride WH, Pajonk F (2006) The response of CD24(-/low)/CD44+ breast cancer-initiating cells to radiation. J Natl Cancer Inst 98:1777–1785PubMedCrossRefGoogle Scholar
  21. Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P et al (2007) Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci USA 104:973–978PubMedCrossRefGoogle Scholar
  22. Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, Henkelman RM, Cusimano MD, Dirks PB (2004) Identification of human brain tumour initiating cells. Nature 432:396–401PubMedCrossRefGoogle Scholar
  23. Takaishi S, Okumura T, Tu S, Wang SS, Shibata W, Vigneshwaran R, Gordon SA, Shimada Y, Wang TC (2009) Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells 27:1006–1020PubMedCrossRefGoogle Scholar
  24. Watkins DN, Berman DM, Burkholder SG, Wang B, Beachy PA, Baylin SB (2003) Hedgehog signalling within airway epithelial progenitors and in small-cell lung cancer. Nature 422:313–317PubMedCrossRefGoogle Scholar
  25. Wu K, Nie Y, Guo C, Chen Y, Ding J, Fan D (2009) Molecular basis of therapeutic approaches to gastric cancer. J Gastroenterol Hepatol 24:37–41PubMedCrossRefGoogle Scholar
  26. Yang ZF, Ho DW, Ng MN, Lau CK, Yu WC, Ngai P et al (2008) Significance of CD90+ cancer stem cells in human liver cancer. Cancer Cell 13:153–166PubMedCrossRefGoogle Scholar
  27. Yu F, Yao H, Zhu P, Zhang X, Pan Q, Gong C, Huang Y, Hu X, Su F, Lieberman J, Song E (2007) let-7 regulates self renewal and tumorigenicity of breast cancer cells. Cell 131:1109–1123PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Chaojun Zhang
    • 1
  • Chenwei Li
    • 2
  • Fengtian He
    • 3
  • Yujiao Cai
    • 1
  • Hua Yang
    • 1
    • 2
  1. 1.Department of General Surgery, Xinqiao HospitalThird Military Medical UniversityChongqingChina
  2. 2.Department of SurgeryUniversity of Michigan Medical CenterAnn ArborUSA
  3. 3.Department of Biochemistry and Molecular BiologyThird Military Medical UniversityChongqingChina

Personalised recommendations