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机构地区:[1]中国医科大学附属第一医院放疗科,辽宁沈阳110001
出 处:《四川医学》2015年第6期799-801,共3页Sichuan Medical Journal
基 金:辽宁省自然科学基金(编号:2013021055)
摘 要:目的研究在常氧状态下HIF-1α对人非霍奇金淋巴瘤(non-Hodgkin lymphoma,NHL)细胞糖酵解途径相关下游靶基因己糖激酶-Ⅱ(hexokinase-Ⅱ,HK-Ⅱ)表达及其对NHL凋亡的调控机制。方法以HIF-1α抑制剂Echinomycin(EC)预处理人NHL细胞及HIF-1α反义质粒转染细胞,检测干预后NHL细胞HK-Ⅱ蛋白表达水平。进一步应用HK-Ⅱ抑制剂Clotrimazole(CLT)预处理人NHL细胞采用Annexin-Ⅴ染色方法检测肿瘤凋亡水平的变化,采用免疫印迹分析方法检测各细胞系中HK-Ⅱ及凋亡相关蛋白cleaved caspase-3的表达水平。结果抑制HIF-1α的表达明显抑制了肿瘤细胞中HK-Ⅱ蛋白表达。HK-Ⅱ抑制剂CLT能够诱导NHL细胞的凋亡(P<0.05),增加cleaved caspase-3蛋白表达。结论常氧状态下HIF-1α通过诱导HK-Ⅱ蛋白表达抑制NHL细胞凋亡。Objective To investigate the role of HIF-1αexpression in control of apoptosis of human non-Hodgkin lympho-ma ( NHL) cells in normoxic condition, as well as to explore the mechanism by inducing hexokinase-Ⅱ( HK-Ⅱ) expression. Methods Three human non-Hodgkin lymphoma cell lines were pre-treated by echinomycin( EC) , a HIF-1αantagonist and pSu-per HIF-1α/siRNA was transfected in NHL cells followed by Western blot for determination of expression level of HK-Ⅱprotein. Then NHL cells were pre-treated by Clotrimazole( CLT) , a HK-Ⅱ antagonist, apoptosis levels were determined with Annexin-Ⅴdying method. Expression levels of apoptotic related protein cleaved caspase-3 were evaluated by Western blot. Results Inhibi-tion of HIF-1α significantly reduced the expression level of HK-Ⅱ.At the same time, CLT significantly induced apoptosis of hu-man NHL cells in a dose dependent manner (P〈0. 05) and reduced the expression level of cleaved caspase-3. Conclusion In-hibition of HIF-1αcould induce cell apoptosis in human NHL cell in normoxic condition through down-regulation of HK-Ⅱexpres-sion.
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