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《Chongqing Medicine》 2017-13
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Experimental study on influence of cytarabine on K562 cells proliferation and apoptosis by autophagy pathway

Luo Hao;Meng Zan;Liu Zehong;Chen Xiaolu;Department of Human Anatomy,Histology and Embryology,Leshan Vocational and Technical College;Department of Pathology,Three Gorges Medical College;Department of Pathology,Leshan Vocational and Technical College;  
Objective To investigate the effect of cytarabine(Ara-C)on proliferation and apoptosis of human erythroleukemia K562 cell line through autophagy pathway and its possible mechanism.Methods The cellular proliferation inhibiting rate after different concentrations of Ara-C acting for 24,48 hwas detected by CCK-8;the cell cycle and apoptosis were detected by flow cytometry(FCM);the chromatin morphological changes in nucleus were observed by Hoechst staining;the cell acidic autophagy vesicles were detected by acridine orange staining;the expression changes of p38 and p-p38 proteins were detected by Western blot.The expressions of autophagy apoptosis related gene and protein were examined by RT-PCR and immunofluorescence.Results The CCK-8results found that different concentrations of Ara-C could inhibit the proliferation of K562 cells with dose-and time-dependent manners.FCM detecting indicated that Ara-C could increase apoptosis and could arrest the cell cycle at S phase;Hoechest staining showed that K562 cells had typical apoptotic morphological changes after Ara-C treating;the Acridine orange staining revealed that Ara-C caused the inclease of the green fluorescene in cells of the Ara-C group,and the cells appeared a great number of acidic autophagy vesicles;RT-PCR results showed that Ara-C up-regulated the expression of autophagy key genes Beclin-1,LC3 Aand LC3B;Western blot results showed that Ara-C increased the expression of phosphorylated p-p38.Immunofluorescence results showed the expression of LC3 Bwas significantly enhanced.Conclusion Ara-C can activate p-p38 mediated K562cells to generate autophagy,then inhibit the cell proliferation and promotes apoptosis.
【CateGory Index】: R96
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