title: Establishment of pancreatic cancer stem cells by flow cytometry and their biological characteristics.
abstract: To investigate the method of separating human pancreatic cancer stem cells by Hoechst 33342 labeled flow cytometry and to analyze the biological properties of pancreatic cancer stem cells. The human pancreatic cancer cell line PC-3 was divided into SP and non-SP cells by flow cytometry. The number of two cell clone spheres and nude mice tumor formation rates were compared by cultivating in serum-free medium; The expression of CD133, Nestin mRNA and protein was analyzed by real-time fluorescence quantitative PCR and Western blot; The expression of two cell drug resistance genes (MDR1, ABCG2, ABCA2 and MRP1) was analyzed by real time fluorescent quantitative PCR. The number of the cloned spheres in SP cells in serum-free medium was significantly higher than that of non-SP cells (P<0.05). The incidence of SP cells in the tumor of immunodeficiency nude mice was significantly higher than that of non-SP cells, and the difference was statistically significant (P<0.05). Real-time fluorescence quantitative PCR analysis showed that the expression of CD133 and Nestin mRNA in SP cells was significantly higher than those of non-SP cells, and the expression of CD133 and Nestin protein in SP cells was also significantly higher than those of non-SP cells (P<0.05). In conclusion, SP side population pancreatic cancer cells by Hoechst 33342 separation have the stem cell characteristics, higher tumor formation rate and higher drug resistance, which may be related to chemotherapy resistance.
authors.affiliation: Department of Endocrinology, Huaihe Hospital of Henan University Kaifeng 475000, Henan, P. R. China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Endocrinology, Huaihe Hospital of Henan University Kaifeng 475000, Henan, P. R. China.
authors.forename: Dongyue
authors.initials: D
authors.lastname: Su
authors.affiliation: Department of Endocrinology, Huaihe Hospital of Henan University Kaifeng 475000, Henan, P. R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xue
authors.affiliation: Department of Endocrinology, Huaihe Hospital of Henan University Kaifeng 475000, Henan, P. R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Endocrinology, Huaihe Hospital of Henan University Kaifeng 475000, Henan, P. R. China.
authors.forename: Wuyan
authors.initials: W
authors.lastname: Pang
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D064231:Nestin / Q000235:genetics / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D058985:Side-Population Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D006023:Glycoproteins; C577484:NES protein, human; D064231:Nestin; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse
keywords: Pancreatic cancer stem cells; SP cells; flow cytometry; non-SP cells