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目的通过克隆形成及致瘤能力研究肺腺癌细胞系A549肿瘤始发细胞的比例。方法采用有限稀释法,检测细胞单克隆形成的数量及大小;收集来自于1个细胞形成的克隆细胞,种于BALB/C裸鼠皮下,观察其致瘤能力;采用CD133磁珠分选A549细胞,测定CD133+、CD133-细胞的克隆形成能力,测定Hoechst33342染色对非SP细胞的增殖能力和单克隆形成能力的影响。结果45%以上的A549细胞能够形成大于100个细胞的大克隆,其传代后又能形成新的克隆,来源于1个大克隆的细胞种植到BALB/C裸鼠身上,10d左右能形成肉眼可见的肿瘤;CD133+、CD133-细胞克隆形成能力相同;Hoechst33342染色能够明显干预细胞的克隆形成及增殖能力。结论45%以上的A549细胞是肿瘤始发细胞。
OBJECTIVE: To study the proportion of tumor-initiating cells in lung adenocarcinoma cell line A549 by clonogenicity and tumorigenicity. Methods The number and size of monoclonal antibodies were detected by limiting dilution method. Clonal cells from one cell line were collected and subcultured in BALB / C nude mice to observe their tumorigenicity. CD133 magnetic beads were used to sort A549 cells , The clonogenic capacity of CD133 + and CD133- cells was measured and the effect of Hoechst33342 staining on the proliferation and monoclonal formation ability of non-SP cells was determined. Results More than 45% of A549 cells could form large clones with more than 100 cells. After passage, new clones could be formed. One large clonal cell was implanted into BALB / C nude mice and could be seen by naked eye about 10 days Of the tumor; CD133 +, CD133-cell clone formation ability of the same; Hoechst33342 staining can significantly interfere with cell cloning formation and proliferation. Conclusions More than 45% of A549 cells are tumor-initiating cells.