OBJECTIVES: Gastric cancer is an important cause of cancer-related mortality worldwide (1). There is increasing evidence that the existence of cancer stem cells (CSC) is responsible for tumour formation and maintenance. MATERIALS AND METHODS: The present study was designed to recognise circulating CSCs from blood samples of patients with gastric cancer, using CD133 and ABCG2 as potential markers. CD133(-) , CD133(+) ABCG2(-) and CD133(+) ABCG2(+) cells lines were analysed by flow cytometry, immunofluorescence staining, western blotting and real-time PCR. Furthermore, functional assays (clonogenic assay in vitro and tumourigenic assay in vivo) were also performed using these cell lines. RESULTS: Higher percentages of CD133(+) cells were identified in blood samples from gastric cancer patients compared to normal controls. In addition, we found by using Kaplan-Meier analysis, that numbers of CD133(+) cells correlated with poor prognosis gastric cancer patients. Finally, tumourigenic properties of CD133(+) ABCG2(+) cells were determined in vitro and in vivo. CONCLUSIONS: Our in vitro and in vivo experiments demonstrated that CD133(+) ABCG2(+) cells exhibited well-known CSC characteristics; thus when circulating they could be used as a prognostic marker for gastric cancer.
OBJECTIVES:Gastric cancer is an important cause of cancer-related mortality worldwide (1). There is increasing evidence that the existence of cancer stem cells (CSC) is responsible for tumour formation and maintenance. MATERIALS AND METHODS: The present study was designed to recognise circulating CSCs from blood samples of patients with gastric cancer, using CD133 and ABCG2 as potential markers. CD133(-) , CD133(+) ABCG2(-) and CD133(+) ABCG2(+) cells lines were analysed by flow cytometry, immunofluorescence staining, western blotting and real-time PCR. Furthermore, functional assays (clonogenic assay in vitro and tumourigenic assay in vivo) were also performed using these cell lines. RESULTS: Higher percentages of CD133(+) cells were identified in blood samples from gastric cancerpatients compared to normal controls. In addition, we found by using Kaplan-Meier analysis, that numbers of CD133(+) cells correlated with poor prognosis gastric cancerpatients. Finally, tumourigenic properties of CD133(+) ABCG2(+) cells were determined in vitro and in vivo. CONCLUSIONS: Our in vitro and in vivo experiments demonstrated that CD133(+) ABCG2(+) cells exhibited well-known CSC characteristics; thus when circulating they could be used as a prognostic marker for gastric cancer.
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