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 ...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 ABCGZ 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.展开更多
Purpose:To study the retinoblastoma cell culture and to establish a new retinoblastoma cellline.Methods:22 retinoblastomes were cultured by using the method of single cellsus-pension.Characteristics of the cultured ce...Purpose:To study the retinoblastoma cell culture and to establish a new retinoblastoma cellline.Methods:22 retinoblastomes were cultured by using the method of single cellsus-pension.Characteristics of the cultured cells were studied in the following pro-grams:tumor cell morphology in vitro,electron microscopic,growth curve,cloning in soft agar,immunohistochemistry,karyotype and tumorigenicity.Results.22 retinoblastoas were cultured successfully in ivtro,only a cotinued cell line HXO-Rb44was established(more than3years).The characteristics of this cell line are that it grew as a suspension of round cell in graps like clusters in vitro,its population doubling time was 44hours,and it could be cloned in softa-gar.Histopathologic and ulatastructured pictures showed the characteristics of Rb.HXO-Rb44cell was positive to NSE and negative to GFAP in immunohis-tochemical staining.A subcutaneous injection of HXO-Rb44cells produced a retinoblastoma in BALB/C athumic nude mice.Conclusions:HXO-Rb44 has the characteristice of retinoblastoma and is a new retinoblastoma cell line.It is a useflu material for study this tumor both in basic and clinical fields.展开更多
AIM: To observe the effects of salvianolic add B (SalB) on in vitro growth inhibition and apoptosis induction of retinoblastoma HXO-RB44 cells. METHODS: The effects of SalB on the HXO-RB44 cells proliferation in vitro...AIM: To observe the effects of salvianolic add B (SalB) on in vitro growth inhibition and apoptosis induction of retinoblastoma HXO-RB44 cells. METHODS: The effects of SalB on the HXO-RB44 cells proliferation in vitro were observed by MTT colorimetric method. The morphological changes of apoptosis before and after the treatment of SalB were observed by Hoechst 33258 fluorescent staining method. Apoptosis rate and cell cycle changes of HXO-RB44 cells were detected by flow cytometer at 48 hours after treated by SalB. The expression changes of Caspase-3 protein in HXO-RB44 cells were detected by Western Blot. RESULTS: SalB significantly inhibited the growth of HXO-RB44 cells, while the inhibition was in a concentration-and time-dependent manner. The results of fluorescent staining method indicated that HXO-RB44 cells showed significant phenomenon of apoptosis including karyorrhexis, fragmentation and the formation of apoptotic bodies, etc. after 24, 48 and 72 hours co-culturing of SalB and HXO-RB44 cells. The results of flow cytometer showed that the apoptosis rate and the proportion of cells in S phase were gradually increased at 48 hours and 72 hours after treated by different concentrations of SalB. Western Blot strip showed that the expression of Caspase-3 protein in HXO-RB44 cells was gradually increased with the increase of the concentration of SalB. CONCLUSION: SalB can significantly affect on HXO-RB44 cells growth inhibition and apoptosis induction which may be achieved through the up-regulation of Caspase-3 expression and the induction of cell cycle arrest.展开更多
基金Supported by National Natural Science Foundation of China(No.81072108No:31050110429)+4 种基金China Postdoctoral Science Foundation(No.20090450167No.201003676)PH.D Programs Foundation of Ministry of Education of China(No. 20090201120068)Scientific and Technological Cooperation Projects of Shaanxi Province,China(2010K14-02)The Fundamental Research Funds for the Central Universities
文摘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 ABCGZ 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.
文摘Purpose:To study the retinoblastoma cell culture and to establish a new retinoblastoma cellline.Methods:22 retinoblastomes were cultured by using the method of single cellsus-pension.Characteristics of the cultured cells were studied in the following pro-grams:tumor cell morphology in vitro,electron microscopic,growth curve,cloning in soft agar,immunohistochemistry,karyotype and tumorigenicity.Results.22 retinoblastoas were cultured successfully in ivtro,only a cotinued cell line HXO-Rb44was established(more than3years).The characteristics of this cell line are that it grew as a suspension of round cell in graps like clusters in vitro,its population doubling time was 44hours,and it could be cloned in softa-gar.Histopathologic and ulatastructured pictures showed the characteristics of Rb.HXO-Rb44cell was positive to NSE and negative to GFAP in immunohis-tochemical staining.A subcutaneous injection of HXO-Rb44cells produced a retinoblastoma in BALB/C athumic nude mice.Conclusions:HXO-Rb44 has the characteristice of retinoblastoma and is a new retinoblastoma cell line.It is a useflu material for study this tumor both in basic and clinical fields.
基金National "Eleventh Five-year Plan" Science and Technology Support Project (No. 2006BAI06 A15-3)
文摘AIM: To observe the effects of salvianolic add B (SalB) on in vitro growth inhibition and apoptosis induction of retinoblastoma HXO-RB44 cells. METHODS: The effects of SalB on the HXO-RB44 cells proliferation in vitro were observed by MTT colorimetric method. The morphological changes of apoptosis before and after the treatment of SalB were observed by Hoechst 33258 fluorescent staining method. Apoptosis rate and cell cycle changes of HXO-RB44 cells were detected by flow cytometer at 48 hours after treated by SalB. The expression changes of Caspase-3 protein in HXO-RB44 cells were detected by Western Blot. RESULTS: SalB significantly inhibited the growth of HXO-RB44 cells, while the inhibition was in a concentration-and time-dependent manner. The results of fluorescent staining method indicated that HXO-RB44 cells showed significant phenomenon of apoptosis including karyorrhexis, fragmentation and the formation of apoptotic bodies, etc. after 24, 48 and 72 hours co-culturing of SalB and HXO-RB44 cells. The results of flow cytometer showed that the apoptosis rate and the proportion of cells in S phase were gradually increased at 48 hours and 72 hours after treated by different concentrations of SalB. Western Blot strip showed that the expression of Caspase-3 protein in HXO-RB44 cells was gradually increased with the increase of the concentration of SalB. CONCLUSION: SalB can significantly affect on HXO-RB44 cells growth inhibition and apoptosis induction which may be achieved through the up-regulation of Caspase-3 expression and the induction of cell cycle arrest.
