中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
Chinese Journal of Pathophysiology
2015年
11期
1928-1932
,共5页
陈谨%周敏然%孙婷%秦雪梅%陈忠敏%陈春燕%于媛
陳謹%週敏然%孫婷%秦雪梅%陳忠敏%陳春燕%于媛
진근%주민연%손정%진설매%진충민%진춘연%우원
三尖杉酯碱%叉头框蛋白M1%K562细胞%药物敏感性
三尖杉酯堿%扠頭框蛋白M1%K562細胞%藥物敏感性
삼첨삼지감%차두광단백M1%K562세포%약물민감성
Homoharringtonine%Forkhead box protein M1%K562 cells%Drug sensitivity
目的：探讨抑制白血病K562细胞叉头框蛋白M1（FoxM1）是否增强细胞对高三尖杉酯碱（HHT）的敏感性。方法：HHT以不同浓度（0、0．015、0．030和0．045μmol／L）和最低起效浓度不同时间（0．015μmol／L，0、24、48和72 h）作用于K562细胞，real－time PCR和Western blot检测FoxM1 mRNA和蛋白表达；以0．015μmol／L HHT作用K562细胞后转染FoxM1 siRNA，观察沉默K562细胞FoxM1后细胞对HHT的敏感性、细胞增殖和凋亡效应以及FoxM1相关靶分子c－Myc和Sp1表达状况。结果：随着HHT浓度增加和时间延长FoxM1表达逐渐降低，说明HHT抑制K562细胞FoxM1表达；HHT处理K562细胞后转染FoxM1 siRNA，细胞生长和克隆形成显著下降，细胞凋亡增加，因此抑制FoxM1可增加K562细胞对HHT的敏感性；FoxM1 siRNA组c－Myc和Sp1表达显著降低，表明FoxM1可正性调控c－Myc和Sp1表达。结论：HHT可以抑制白血病K562细胞FoxM1表达，干扰FoxM1可增强细胞对HHT的敏感性。
目的：探討抑製白血病K562細胞扠頭框蛋白M1（FoxM1）是否增彊細胞對高三尖杉酯堿（HHT）的敏感性。方法：HHT以不同濃度（0、0．015、0．030和0．045μmol／L）和最低起效濃度不同時間（0．015μmol／L，0、24、48和72 h）作用于K562細胞，real－time PCR和Western blot檢測FoxM1 mRNA和蛋白錶達；以0．015μmol／L HHT作用K562細胞後轉染FoxM1 siRNA，觀察沉默K562細胞FoxM1後細胞對HHT的敏感性、細胞增殖和凋亡效應以及FoxM1相關靶分子c－Myc和Sp1錶達狀況。結果：隨著HHT濃度增加和時間延長FoxM1錶達逐漸降低，說明HHT抑製K562細胞FoxM1錶達；HHT處理K562細胞後轉染FoxM1 siRNA，細胞生長和剋隆形成顯著下降，細胞凋亡增加，因此抑製FoxM1可增加K562細胞對HHT的敏感性；FoxM1 siRNA組c－Myc和Sp1錶達顯著降低，錶明FoxM1可正性調控c－Myc和Sp1錶達。結論：HHT可以抑製白血病K562細胞FoxM1錶達，榦擾FoxM1可增彊細胞對HHT的敏感性。
목적：탐토억제백혈병K562세포차두광단백M1（FoxM1）시부증강세포대고삼첨삼지감（HHT）적민감성。방법：HHT이불동농도（0、0．015、0．030화0．045μmol／L）화최저기효농도불동시간（0．015μmol／L，0、24、48화72 h）작용우K562세포，real－time PCR화Western blot검측FoxM1 mRNA화단백표체；이0．015μmol／L HHT작용K562세포후전염FoxM1 siRNA，관찰침묵K562세포FoxM1후세포대HHT적민감성、세포증식화조망효응이급FoxM1상관파분자c－Myc화Sp1표체상황。결과：수착HHT농도증가화시간연장FoxM1표체축점강저，설명HHT억제K562세포FoxM1표체；HHT처리K562세포후전염FoxM1 siRNA，세포생장화극륭형성현저하강，세포조망증가，인차억제FoxM1가증가K562세포대HHT적민감성；FoxM1 siRNA조c－Myc화Sp1표체현저강저，표명FoxM1가정성조공c－Myc화Sp1표체。결론：HHT가이억제백혈병K562세포FoxM1표체，간우FoxM1가증강세포대HHT적민감성。
AIM:To study whether inhibition of forkhead box protein M1(FoxM1) sensitizes leukemia K562 cells to homoharringtonine ( HHT ) .METHODS: K562 cells were incubated with HHT at different concentrations ( 0μmol/L, 0.015 μmol/L, 0.030μmol/L and 0.045μmol/L) for different time (0 h, 24 h, 48 h and 72 h).The mRNA and protein levels of FoxM1 were detected by real-time PCR and Western blot.FoxM1 siRNA was transfected into K562 cells with 0.015μmol/L HHT after 6 h.After 72 h incubation, the cell proliferation was detected by cell counting and soft agar assay, and the proportion of apoptotic K562 cells was determined by flow cytometry.The expression of c-Myc and Sp1 were detected by real-time PCR and Western blot.RESULTS:FoxM1 expression was reduced time-dependently and dose-dependently, suggesting that HHT mediated the downregulation of FoxM1 in K562 cells.In K562 cells, treatment with FoxM1 siRNA and HHT inhibited the cell proliferation and promoted the apoptosis significantly.Therefore, inhibition of FoxM1 sensitized leukemia K562 cells to HHT.The expression of c-Myc and Sp1 was positively regulated by FoxM1. CONCLUSION:HHT inhibits Forkhead box protein M1 expression in K562 cells.Inhibition of FoxM1 sensitizes K562 cells to HHT.
