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目的进一步阐明白血病细胞对伊马替尼耐药的机制。方法经过对K562细胞系长期的鬼臼乙叉甙(Vp16)诱导和克隆筛选,建立了一株耐药细胞系K562/Vp16,利用干细胞高效能将Hoechst33342荧光染料泵出细胞的特性,采用流式细胞术从K562/Vp16细胞系中分选出一小群细胞,即边缘细胞(Side population,SP),称为K562/Vp16SP细胞,并初步探讨了其对伊马替尼耐药的机制。结果BCR/ABL和ABL蛋白在K562细胞、K562/Vp16SP细胞及K562/Vp16非SP细胞(K562/Vp16non-SP)中的表达水平差异无统计学意义;P-gp在K562细胞中不表达,在K562/Vp16SP及K562/Vp16non-SP细胞中均高表达且表达水平一致;与K562/Vp16non-SP细胞比较,K562/Vp16SP细胞对伊马替尼的耐药性更强,并且这种耐药性几乎不能被多种多药耐药逆转剂逆转;另外,体内外实验显示,K562/Vp16细胞的致瘤性几乎全部来源于K562/Vp16SP细胞。结论白血病细胞对伊马替尼具有一定的耐药性,可能与数量极少的SP细胞有直接的关系。因此,这类数量极少的SP细胞应当成为有效治疗肿瘤的靶细胞。
Objective To further elucidate the mechanism of resistance of leukemic cells to imatinib. Methods A multidrug-resistant cell line K562 / Vp16 was induced and cloned by long-term stimulation of Vp16 in K562 cell line. The characteristics of Hoechst33342 fluorescent dye were pumped out of cells with high efficiency stem cells. A small group of cells (K562 / Vp16SP cells) were sorted out from the K562 / Vp16 cell line by cytometry. The mechanism of resistance to imatinib was also discussed. Results There was no significant difference in the expression of BCR / ABL and ABL protein between K562 cells, K562 / Vp16SP cells and K562 / Vp16 non-SP cells (K562 / Vp16non-SP); P-gp was not expressed in K562 cells. K562 / Vp16SPA and K562 / Vp16non-SP cells. The K562 / Vp16SP cells were more resistant to imatinib than K562 / Vp16non-SP cells, and the drug resistance Almost can not be reversed by a variety of multidrug resistance reversal agents; In addition, in vitro and in vivo experiments show that, K562 / Vp16 cell tumorigenic almost entirely from K562 / Vp16SP cells. Conclusion Leukemic cells have some resistance to imatinib, which may be directly related to a small number of SP cells. Therefore, such a very small number of SP cells should be the target cells for the effective treatment of tumors.