Side population cells from HXO-Rb44 retinoblastoma cell line have cancer-initiating property.
Side population cells from HXO-Rb44 retinoblastoma cell line have cancer-initiating property.
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DOI：
10.3980/j.issn.2222-3959.2011.05.01
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发表时间：
2011
影响因子：
1.4
通讯作者：
J. She;Peng-Ge Zhang;X. Che;Xuan Wang;Zi-Ming Wang
中科院分区：
文献类型：
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作者：
J. She;Peng-Ge Zhang;X. Che;Xuan Wang;Zi-Ming Wang
目的 确定HXO-Rb44视网膜母细胞瘤细胞系中的侧群（SP）细胞在体外和体内是否具有癌症干细胞样特性。方法 我们通过流式细胞仪上的 Hoechst 33342 染色对来自 HXO-Rb44 视网膜母细胞瘤细胞系的 SP 进行分析和分选。通过SP再分析、软琼脂实验和体外肿瘤球实验测定SP和NSP细胞的增殖和自我更新能力。通过将 HXO-Rb44 和 HXO-Rb44 -RFP 细胞接种到软琼脂中来检测克隆形成。通过 RT-PCR 测定 SP 和非 SP (NSP) 细胞之间 ABCG2、MDRI、Bmi-1 和 Oct-4 的表达。此外，将它们注射到裸鼠体内以确定其体内肿瘤性。结果 来自 HXO-Rb44 视网膜母细胞瘤细胞系的 SP 可以在软琼脂测定中克隆生长，并在条件培养基中从单细胞形成肿瘤球。 SP细胞中ABCG2、MDRI、Bmi-1和Oct-4的表达显着高于NSP细胞。 12个注射部位中，只有少数SP细胞导致6个部位形成肿瘤，然而，注射NSP细胞却未能形成新的肿瘤。结论 Hoechst 33342从HXO-Rb44视网膜母细胞瘤细胞系中分离得到的SP细胞在体内外均具有高致瘤性。因此，SP可能是开发视网膜母细胞瘤疗法的一个靶点。
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.