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AIM:To ascertain whether side population(SP) cells in HXO-Rb44 retinoblastoma cell line have cancer stem cell-like property in vitro and in vivo.METHODS:We analyzed and sorted SP from HXO-Rb44 retinoblastoma cell line by Hoechst 33342 staining on flow cytometry.SP and NSP cells were determined their ability of proliferation and self-renewal by SP reanalysis,soft agar assay and tumor sphere assay in vitro.Clone formation was detected by seeding HXO-Rb44 and HXO-Rb44-RFP cells into soft agar.The expression of ABCG2,MDRI,Bmi-1 and Oct-4 was determined by RT-PCR between SP and non-SP(NSP) cells.Moreover,they were injected into nude mice to determine their tumorigency in vivo.RESULTS:SP from HXO-Rb44 retinoblastoma cell line could grow clonally in soft agar assays and form tumor spheres from single cells in conditioned media.The expressions of ABCG2,MDRI,Bmi-1 and Oct-4 were significantly higher in SP than NSP cells.As few as SP cells resulted in tumor formation in 6 of 12 injected sites,however,the injection of NSP cells failed to form new tumor.CONCLUSION:SP cells isolated by Hoechst 33342 from the HXO-Rb44 retinoblastoma cell line had property of high tumorigency in vivo and in vitro.Therefore,SP might be a target while developing retinoblastoma therapies.
AIM: To ascertain whether side population (SP) cells in HXO-Rb44 retinoblastoma cell line have cancer stem cell-like property in vitro and in vivo. METHODS: We analyzed and sorted SP from HXO-Rb44 retinoblastoma cell line by Hoechst 33342 staining on flow cytometry. SP and NSP cells were determined their ability of proliferation and self-renewal by SP reanalysis, soft agar assay and tumor sphere assay in vitro. Clone formation was detected by seeding HXO-Rb44 and HXO-Rb44-RFP cells into soft agar The expression of ABCG2, MDRI, Bmi-1 and Oct-4 was determined by RT-PCR between SP and non-SP (NSP) cells. Moreover, they were injected into nude mice to determine their tumorigency in vivo. from HXO-Rb44 retinoblastoma cell line could grow clonally in soft agar assays and form tumor spheres from single cells in conditioned medium. These expressions of ABCG2, MDRI, Bmi-1 and Oct-4 were significantly higher in SP than NSP cells. as SP cells resulted in tumor formation in 6 of 12 injected sites, ho wever, the injection of NSP cells failed to form new tumor. CONCLUSION: SP cells isolated by Hoechst 33342 from the HXO-Rb44 retinoblastoma cell line had property of high tumorigency in vivo and in vitro.Therefore, SP might be a target while developing retinoblastoma therapies.