Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
권호 표지
DOI QR코드

DOI QR Code

Establishment of a Pancreatic Cancer Stem Cell Model Using the SW1990 Human Pancreatic Cancer Cell Line in Nude Mice

  • Pan, Yan (Department of Integrative Oncology, Fudan University Shanghai Cancer Center) ;
  • Gao, Song (Department of Integrative Oncology, Fudan University Shanghai Cancer Center) ;
  • Hua, Yong-Qiang (Department of Integrative Oncology, Fudan University Shanghai Cancer Center) ;
  • Liu, Lu-Ming (Department of Integrative Oncology, Fudan University Shanghai Cancer Center)
  • Published : 2015.02.25
Download PDF
⟨ Previous Next ⟩

Abstract

Aim: To establish a pancreatic cancer stem cell model using human pancreatic cancer cells in nude mice to provide a platform for pancreatic cancer stem cell research. Materials and Methods: To establish pancreatic cancer xenografts using human pancreatic cancer cell line SW1990, nude mice were randomly divided into control and gemcitabine groups. When the tumor grew to a volume of $125mm^3$, they treated with gemcitabine at a dose of 50mg/kg by intraperitoneal injection of 0.2ml in the gemcitabine group, while the mice in control group were treated with the same volume of normal saline. Gemcitabine was given 2 times a week for 3 times. When the model was established, the proliferation of pancreatic cancer stem cells was observed by clone formation assay, and the protein and/or mRNA expression of pancreatic stem cell surface markers including CD24, CD44, CD133, ALDH, transcription factors containing Oct-4, Sox-2, Nanog and Gli, the key nuclear transcription factor in Sonic Hedgehog signaling pathway was detected by Western blot and/or RT-PCR to verify the reliability of this model. Results: This model is feasible and safe. During the establishment, no mice died and the weight of nude mice maintained above 16.5g. The clone forming ability in gemcitabine group was stronger than that of the control group (p<0.01). In gemcitabine group, the protein expression of pancreatic cancer stem cell surface markers including CD44, and ALDH was up-regulated, the protein and mRNA expression of nuclear transcription factor including Oct-4, Sox-2 and Nanog was also significantly increased (P<0.01). In addition, the protein expression of key nuclear transcription factor in Sonic Hedgehog signaling pathway, Gli-1, was significantly enhanced (p<0.01). Conclusions: The pancreatic cancer stem cell model was successfully established using human pancreatic cancer cell line SW1990 in nude mice. Gemcitabine could enrich pancreatic cancer stem cells, simultaneously accompanied by the activation of Sonic Hedgehog signaling pathway.

Keywords

References

  1. Asuthkar S, Stepanova V, Lebedeva T, et al (2013). Multifunctional roles of urokinase plasminogen activator (uPA) in cancer stemness and chemoresistance of pancreatic cancer. Mol Biol Cell, 24, 2620-32. https://doi.org/10.1091/mbc.E12-04-0306
  2. Clarke MF, Dick JE, Dirks PB, et al (2006). Cancer stem cells--perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res, 66, 9339-44. https://doi.org/10.1158/0008-5472.CAN-06-3126
  3. Canyilmaz E, Serdar L, Uslu GH, et al (2013). Evaluation of prognostic factors and survival results in pancreatic carcinomas in Turkey. Asian Pac J Cancer Prev, 14, 6573-8. https://doi.org/10.7314/APJCP.2013.14.11.6573
  4. Dalerba P, Cho RW, Clarke MF (2007). Cancer stem cells: models and concepts. Annu Rev Med, 58, 267-84. https://doi.org/10.1146/annurev.med.58.062105.204854
  5. Gach PC, Attayek PJ, Herrera G, et al (2013). Isolation and in vitro culture of rare cancer stem cells from patient-derived xenografts of pancreatic ductal adeno-carcinoma. Anal Chem, 85, 7271-78. https://doi.org/10.1021/ac401165s
  6. Hermann PC, Huber SL, Herrler T, et al (2007). Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell, 1, 313-23. https://doi.org/10.1016/j.stem.2007.06.002
  7. Jimeno A, Feldmann G, Suarez-Gauthier A et al (2009). A direct pancreatic cancer xenograft model as a platform for cancer stem cell therapeutic development. Mol Cancer Ther, 8, 310-4. https://doi.org/10.1158/1535-7163.MCT-08-0924
  8. Kim HS, Yoo SY, Kim KT, et al (2012). Expression of the stem cell markers CD133 and nestin in pancreatic ductal adenocarcinoma and clinical relevance. Int J Clin Exp Pathol, 5, 754-61.
  9. Li C, Heidt DG, Dalerba P et al (2007). Identification of pancreatic cancer stem cellsJ. Cancer Res, 67, 1030-7. https://doi.org/10.1158/0008-5472.CAN-06-2030
  10. Lu Y, Zhu H, Shan H, et al (2013). Knockdown of Oct4 and Nanog expression inhibits the stemness of pancreatic cancer cells. Cancer Lett, 340, 113-23. https://doi.org/10.1016/j.canlet.2013.07.009
  11. Ohara Y, Oda T, Sugano M, et al (2013). Histological and prognostic importance of CD44 (+)/CD24 (+)/Ep CAM (+) expression in clinical pancreatic cancer. Cancer Sci, 104, 1127-34. https://doi.org/10.1111/cas.12198
  12. Pan Y, Hua YQ, Liu LM, et al (2012). Isolation and identification of pancreatic cancer stem cells from BxPC-3 cell line cultured in serum-free medium. China Cancer, 21, 441-5.
  13. Rasheed ZA, Matsui W (2012). Biological and clinical relevance of stem cells in pancreatic adenocarcinoma. Gastroenterol Hepatol, 27, 15-8. https://doi.org/10.1111/j.1440-1746.2011.07015.x
  14. Rasheed ZA, Yang J, Wang Q, et al (2010). Prognostic significance of tumorigenic cells with mesenchymal features in pancreatic adenocarcinoma. Natl Cancer Inst, 102, 340-51. https://doi.org/10.1093/jnci/djp535
  15. Trumpp A, Wiestler OD (2008). Mechanisms of Disease: cancer stem cells--targeting the evil twin. Nat Clin Pract Oncol, 5, 337-47.
  16. Van den Broeck A, Gremeaux L, Topal B, et al (2012). Human pancreatic adenocarcinoma contains a side population resistant to gemcitabine. BMC Cancer, 12, 354. https://doi.org/10.1186/1471-2407-12-354
  17. Wang X, Liu Q, Hou B, et al (2013). Concomitant targeting of multiple key transcription factors effectively disrupts cancer stem cells enriched in side population of human pancreatic cancer cells. PLoS One, 8, 73942. https://doi.org/10.1371/journal.pone.0073942
  18. Yin T, Wei H, Gou S, et al (2011). Cancer stem-like cells enriched in panc-1 spheres possess increased migration ability and resistance to gemcitabine. Int J Mol Sci, 12, 1595-604. https://doi.org/10.3390/ijms12031595
  19. Zhou BB, Zhang H, Damelin M, et al (2009). Tumour-initiating cells: challenges and opportunities for anticancer drug discovery. Nat Rev Drug Discov, 8, 806-23. https://doi.org/10.1038/nrd2137