四川医学
四川醫學
사천의학
SICHUAN MEDICAL JOURNAL
2015年
6期
799-801
,共3页
淋巴瘤%HIF-1α%HXKⅡ%凋亡
淋巴瘤%HIF-1α%HXKⅡ%凋亡
림파류%HIF-1α%HXKⅡ%조망
Lymphoma%HIF-1α%HXKⅡ%Apoptosis
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Echino-mycin( 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。
目的:研究在常氧狀態下HIF-1α對人非霍奇金淋巴瘤( non-Hodgkin lymphoma, NHL)細胞糖酵解途徑相關下遊靶基因己糖激酶-Ⅱ( hexokinase-Ⅱ,HK-Ⅱ)錶達及其對NHL凋亡的調控機製。方法以HIF-1α抑製劑Echino-mycin( EC)預處理人NHL細胞及HIF-1α反義質粒轉染細胞,檢測榦預後NHL細胞HK-Ⅱ蛋白錶達水平。進一步應用HK-Ⅱ抑製劑Clotrimazole( CLT)預處理人NHL細胞採用Annexin-Ⅴ染色方法檢測腫瘤凋亡水平的變化,採用免疫印跡分析方法檢測各細胞繫中HK-Ⅱ及凋亡相關蛋白cleaved caspase-3的錶達水平。結果抑製HIF-1α的錶達明顯抑製瞭腫瘤細胞中HK-Ⅱ蛋白錶達。 HK-Ⅱ抑製劑CLT能夠誘導NHL細胞的凋亡( P<0.05),增加cleaved caspase-3蛋白錶達。結論常氧狀態下HIF-1α通過誘導HK-Ⅱ蛋白錶達抑製NHL細胞凋亡。
목적:연구재상양상태하HIF-1α대인비곽기금림파류( non-Hodgkin lymphoma, NHL)세포당효해도경상관하유파기인기당격매-Ⅱ( hexokinase-Ⅱ,HK-Ⅱ)표체급기대NHL조망적조공궤제。방법이HIF-1α억제제Echino-mycin( EC)예처리인NHL세포급HIF-1α반의질립전염세포,검측간예후NHL세포HK-Ⅱ단백표체수평。진일보응용HK-Ⅱ억제제Clotrimazole( CLT)예처리인NHL세포채용Annexin-Ⅴ염색방법검측종류조망수평적변화,채용면역인적분석방법검측각세포계중HK-Ⅱ급조망상관단백cleaved caspase-3적표체수평。결과억제HIF-1α적표체명현억제료종류세포중HK-Ⅱ단백표체。 HK-Ⅱ억제제CLT능구유도NHL세포적조망( P<0.05),증가cleaved caspase-3단백표체。결론상양상태하HIF-1α통과유도HK-Ⅱ단백표체억제NHL세포조망。
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.