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背景与目的:近几年研究发现,CRKL(CT10regulatorofkinaselike)基因编码的蛋白在慢性粒细胞白血病(chronicmyeloidleukemia,CML)发病中起重要作用。本实验研究CRKL基因反义寡核苷酸对慢性粒细胞白血病细胞系K562细胞凋亡的影响。方法:以脂质体为载体,用CRKL基因反义寡核苷酸转染K562细胞,通过活细胞计数、透射电镜、流式细胞术、DNALadder测定观察K562细胞形态学、细胞周期、基因表达等的变化。结果:在CRKL基因反义寡核苷酸作用下,K562细胞的生长明显出现抑制;流式细胞仪检测见二倍体峰前出现凋亡峰;在电镜下可见处于凋亡各期的K562细胞;提取其基因组DNA进行琼脂糖凝胶电泳,可见明显的细胞凋亡梯状条带。而对照组K562细胞未观察到上述变化。结论:CRKL基因是Ph染色体阳性CML发病的重要因素。
BACKGROUND & AIM: In recent years, it has been found that the protein encoded by CRKL (CT10 regulator ofkinaselike) plays an important role in the pathogenesis of chronic myeloid leukemia (CML). This study was designed to investigate the effects of CRKL antisense oligonucleotide on apoptosis of chronic myeloid leukemia cell line K562. METHODS: K562 cells were transfected with CRKL antisense oligonucleotides using liposome as carrier. The morphology, cell cycle and gene expression of K562 cells were observed by viable cell counting, transmission electron microscopy, flow cytometry and DNALadder assay The change. Results: Under the action of CRKL antisense oligonucleotide, the growth of K562 cells was obviously inhibited. The apoptosis peak appeared before the peak of diploid by flow cytometry. The apoptosis of K562 cells The genomic DNA was extracted and subjected to agarose gel electrophoresis. A clear ladder of apoptotic cells was observed. The control group K562 cells did not observe the above changes. Conclusion: The CRKL gene is an important factor in the pathogenesis of Ph chromosome positive CML.