Establishment of a pancreatic cancer stem cell model using the SW1990 human pancreatic cancer cell line in nude mice.

Yan Pan,Song Gao,Yong-Qiang Hua,Lu-Ming Liu

doi:10.7314/apjcp.2015.16.2.437

Abstract

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. 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 125 mm3, they treated with gemcitabine at a dose of 50 mg/kg by intraperitoneal injection of 0.2 ml 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. This model is feasible and safe. During the establishment, no mice died and the weight of nude mice maintained above 16.5 g. 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). 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.

Highlights

Pancreatic Cancer Stem Cell (PCSC) represents the tumor’s subpopulation with the capacity to drive its growth, invasion and metastasis, and severely affects the clinical prognosis (Clarke et al, 2006; Dalerba et al, 2007)
The results indicated that the protein expression of pancreatic cancer stem cell surface markers increased in different degree. the protein expression of CD44, The identification of Human SW1990 PCSC
aldehyde dehydrogenase (ALDH) is significantly up-regulated (t test, p=0.000), After the gemcitabine induction, we identified the while Membrane CD24 and CD133 did not significantly enrichment condition of PCSC in gemcitabine group by increased between groups and had no statistical variation detecting the followings: 1) Human PCSC proliferation; compared with the control group (p=0.064, p=0.053). (

Summary

Introduction

Pancreatic Cancer Stem Cell (PCSC) represents the tumor’s subpopulation with the capacity to drive its growth, invasion and metastasis, and severely affects the clinical prognosis (Clarke et al, 2006; Dalerba et al, 2007). PCSC was discovered for the first time in 2007 (Li et al, 2007). Emerging evidence has suggested that PCSC serve as a reservoir for chemoresistence, recurrence and tumor metastasis. It would benefit for the treatment and the patient survive through PCSC -directed therapies (Trumpp et al, 2008; Zhou et al, 2009). The key step is to establish a scientific and feasible animal model for the study on mechanism of PCSC as well as the screening of effective medicines. The acquisition of PCSC and the foundation of animal model haven’t had an acknowledged solution

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