阿糖胞苷通过抑制HDAC6促进白血病细胞K562凋亡的机制研究  

Study on the Mechanism of Cytarabine Promoting Apoptosis of Leukemia Cells K562 by Inhibiting HDAC6

作　　者：罗昊[1] 陈玉[2] 谭丽[3] 孟赞[1] 陈晓露[2] Luo Hao;Chen Yu;Tan Li;Meng Zan;Chen Xiaolu(Department of Histology and Embryology, Leshan Vocational ＆ Technical college, Leshan, 614000;Department of Pathology, Leshan Vocationaland ＆ Technical College, Leshan, 614000;Department of Surgery, Leshan Vocational ＆ Technical college, Leshan, 614000)

机构地区：[1]乐山职业技术学院人体解剖学与组织胚胎学教研室,乐山614000 [2]乐山职业技术学院病理学教研室,乐山614000 [3]乐山职业技术学院外科教研室,乐山614000

出　　处：《基因组学与应用生物学》2018年第7期3137-3142,共6页Genomics and Applied Biology

基　　金：乐山职业技术学院资助

摘　　要：为了探讨阿糖胞苷（Ara-C）通过组蛋白乙酰化酶6（HDAC6）影响人红白血病K562细胞株凋亡作用及可能的机制。采用CCK-8法检测不同浓度的Ara-C作用24h后检测细胞活力;流式细胞术（FCM）检测细胞凋亡率;Hoechest染色观察细胞核染色质的形态;RT-PCR检测HDAC1-6基因的表达变化;Western blotting检测HDAC6、p38和p-p38的蛋白表达。CCK-8检测显示不同浓度的Ara-C能抑制K562细胞的活力,并呈浓度依赖性;FCM检测显示Ara-C能增加细胞的凋亡率;Hoechest染色发现Ara-C组细胞呈凋亡形态学改变;RT-PCR检测显示Ara-C能降低HDAC1、HDAC2和HDAC6的表达;FCM和Hoechest染色发现HDAC抑制能增强Ara-C诱导K562细胞凋亡;Western blotting检测发现Ara-C能降低HDAC6,增加磷酸化p38和激活型Caspase-3表达。可见Ara-C能够通过抑制HDAC6激活p38,诱导K562细胞发生凋亡。In order to investigate the effect of Ara-C on proliferation and apoptosis of human erthroleukemia K562 cell line through Histone Deacetylases 6 （HDAC6） pathway as well as the relevant mechanism. The test groups were incubated with Ara-C at various concentrations for 24 h were determined for proliferative activity by CCK-8; The changes of cell cycle apoptosis were detected by Flow cytometry （FCM）; The cell morphological changes of apoptosis were observed by Hoechst staining; The expressions of HDA C1-6 gene were examined by RT-PCR; The expression of HDAC6, p38 and p-p38 proteins were detected by Western blotting. Ara-C could inhibit the proliferation of K562 cells significantly with dose dependent manners; FCM indicated that Ara-C induced apoptosis; Hoechest staining showed that K562 cells had typical apoptotic morphological changes by treated Ara-C; RT-PCR results show that Ara-C decreased the expression of HDAC 1, HDAC2 and HDAC6; Flow cytometry and Hoechest staining showed that HDAC inhibitors enhanced Ara-C induced apoptosis in K562 cells; Western blot results showed the expression of HDAC6 was reduced, p-p38 and activated Caspase-3 was significantly enhanced. Conclusion： Ara-C significantly inhibited the proliferation and induced apoptosis through inhibiting HDAC6 activated p-p38 in K562 cells.

关 键 词：阿糖胞苷 白血病 蛋白乙酰化酶6 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]