PKC-ε、cIAP在CD44介导的髓系白血病细胞转移和增殖中的作用及可能机制  

Study on the metastasis and proliferation of myelocytic leukemic cells mediated by PKC-ε regulating cIAP and CD44

作　　者：毕作木 崔春萍 孙桂珍 徐冬[1] 周其锋[1] 

机构地区：[1]潍坊医学院附属淄博市第一医院血液科,山东淄博255200 [2]中国人民解放军军事医学科学院放射与辐射医学研究所血液学研究室,北京100039 [3]淄博市沂源县人民医院,山东淄博255200

出　　处：《中国生化药物杂志》2014年第6期1-5,10,共6页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金青年科学基金

摘　　要：目的 探讨PKC-ε在调节cIAP、CD44及P-gp介导的髓系白血病细胞转移、增殖及耐药中的作用及可能机制.方法 采用MTT法检测白血病细胞亲本株及耐药株K562、K562R、HL60和HL60R的耐药特性;Western blot和免疫组化检测各细胞株中PKC-ε表达水平;免疫荧光和流式细胞术检测各细胞株CD44表达;软琼脂克隆形成实验检测细胞株增殖能力;免疫荧光检测PKC-ε与P-gp、cIAP和CD44共定位.结果 髓系白血病耐药株较亲本细胞株高表达PKC-ε,采用化疗药物处理可上调亲本株PKC-ε的表达.通过添加PKC激动剂或抑制剂证实:髓系白血病细胞的转移及耐药与PKC-ε的高表达有关,增殖与PKC-ε的低表达有关.细胞及转移灶中,cIAP及P-gp的表达与PKC-ε的表达变化一致,CD44的表达与PKC-ε的表达变化相反.免疫荧光结果显示PKC-ε与P-gp、cIAP及CD44分别共定位于髓系白血病细胞膜上.结论 PKC-ε通过调节P-gp和cIAP介导白血病细胞转移,同时参与调节CD44介导的白血病细胞增殖.Objective To explore the mechanism of PKC-ε though cIAP,CD44 and P-gp on myeloid leukemia cell transfer,proliferation and drug resistance. Methods The resistant characteristics of leukemia cell parents lines and drug-resistant strains K562,K562 R,HL60 and HL60 R were determined by MTT method; PKC-ε expression levels in each strains were detected by Western blot and immunohistochemistry; CD44 expression in each strains were detected by immunofluorescence and flow cytomery; the proliferation ability of cell lines were detected by soft agarose assay; co-location of PKC-ε and P-gp,cIAP and CD44 were studied by immunofluorescence. Results The experimental results shoued that the expression of PKC-ε in the myeloid leukemia drug-resistant strains were higher than the parent cell lines,and chemotherapy drug treatment could increase the PKC-ε expression of parent strains. PKC agonist and inhibition test confirmed that the correlation between myeloid leukemia cell transfer and drug resistance and PKC-εexpression was positive,and the correlation between myeloid leukemia cell proliferation and PKC-ε expression was negative. Cells and metastases,cIAP and P-gp expression and the expression of PKC-ε was positively correlated,and the expression of CD44 was negatively correlated to the expression of PKC-ε. Immunofluorescence results showed that PKC-ε and P-gp, cIAP and CD44 were co-located in myeloid leukemia cell membranes.Conclusion PKC-ε mediate leukemic cell transfer though regulating P-gp and cIAP,while regulate the proliferation of leukemic cell mediated by CD44.

关 键 词：PKC-ε cIAP 髓系白血病细胞 增殖 转移 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]