裸鼠高成瘤性多药耐药白血病细胞系的建立及其生物学特性研究

目的:建立裸鼠高成瘤性多药耐药白血病细胞系并研究其生物学特性.方法:采用逐步递增长春新碱(VCR)剂量的方法培养高成瘤性人白血病细胞系K562-n,采用细胞培养技术、透射电镜观察、流式细胞术、RT-PCR、免疫组化、染色体核型分析及体内接种等方法研究其生物学特性.结果:建立1株裸鼠高成瘤性人白血病多药耐药细胞系K562-n/VCR.与K562-n细胞比较,K562-n/VCR细胞对VCR耐药为其297.38倍,对蒽环类、鬼臼类等多种化疗药物具有交叉耐药性,bcr-abl融合基因仍为阳性,裸鼠体内成瘤性不变,但成瘤潜伏期缩短,成瘤体积明显增大.结论:裸鼠高成瘤性多药耐药白血病细胞系K562-n/VCR具有其独特的生物学特性.

Establishment and characterization of a multi-drug resistant leukemia cell line with high tumorigenicity innude mice

Objective:To establish a multi-drug resistant leukemia cell line (K562-n/VCR) with high tumorigenicity in nude mice and to study its biological features. Methods : K562-n cells were induced into multi-drug resistant cell line K562-n/ VCR by a progressively increasing doses of VCR. The biological features of K562-n/VCR,including cell growth,tumorigenicity in nude mice,colony formation percentage in soft agar, ultrastructure under electron microscopy,cell cycle distribution and the intracellular accumulation of DNR by flow cytometer ,were compared with those of K562-n. The cytogenetics analysis of different strains was also done to observe the chromosome karotype. MTT method was used to determine the cytotoxity of some anticancer drugs. Expression of p-gp was detected by immunohistochemical method. RT-PCR was applied for detection of mdr-1 gene expression. Results: K562-n/VCR cells were found to be cross-resistant to VCR,DNR,EPI,HHT,NVT,VP16 and VM26. Compared with K562-n cells,the resistance of K562-n/VCR to VCR was 297. 38 folds. There was no significantly difference of tumorigenicity in nude mice between K562-n/VCR and K562-n cell line. But the latent period of the tumor was much shorter and volume of mass was bigger in nude mice inoculated K562-n/VCR cells. The bcr-abl gene was still positive in K562-n/VCR cell lines. Conclusion: A multi-drug resistant leukemia cell line K562-n/VCR, with high tumorigenicity and u-nique biological feature,had been established.