5-脂氧合酶激活蛋白抑制剂MK886对人食管癌细胞系增殖和凋亡的影响

Effects of 5-lipoxygenase activating protein inhibitor MK886 on proliferation and apoptosis of human esophageal cancer cell lines

目的5-脂氧合酶活化蛋白(FLAP)抑制剂MK886对人食管癌细胞系(KYSE-150和TE-3)增殖和凋亡的影响及作用机制。方法用2.5、5、10、20、40和80μmol/L的MK886干预体外培养的KYSE-150和TE-3细胞;xCELLigence RTCA系统实时测定细胞增殖抑制率,同时确定半数抑制浓度(IC_(50))。流式细胞测量术检测食管癌细胞的细胞周期。Western blot检测细胞凋亡和自噬相关蛋白的表达。结果人食管癌细胞系(KYSE-150和TE-3)的增殖抑制率随着MK886浓度增加而增强(P<0.05),KYSE-150组的IC_(50)浓度为29.11μmol/L,TE-3组的IC_(50)浓度为27.47μmol/L。当MK886浓度增加至25μmol/L时,食管癌细胞G0/G1期滞后增加明显(P<0.001),MK886处理浓度增加到50μmol/L时,食管癌细胞G2/M期增加明显(P<0.001和P<0.05)。MK886处理后食管癌细胞系随着浓度增加,cleaved-caspase-3和LC-3A/B-Ⅱ蛋白表达水平增强(P<0.001)。结论MK886能抑制人食管癌细胞系的增殖,引起凋亡,并影响细胞周期变化。

Objective To investigate the effects of 5-lipoxygenase activating protein(FLAP)inhibitor MK886 on the proliferation and apoptosis of human esophageal carcinoma cell lines(KYSE-150 and TE-3)and to explore the potential mechanism.Methods KYSE-150 and TE-3 cells were treated with MK886(2.5,5,10,20,40 and 80μmol/L).xCELLigence RTCA system was used to to detect the half maximal inhibitory concentration(IC_(50))of MK886.The expressions of proteins with apoptosis and autophagy were detected by Western blot and cell cycles were detected by flow cytometry.Results MK886 decreased the proliferation of human esophageal carcinoma cells(KYSE-150 and TE-3)(both P<0.05).IC_(50) of MK886 in KYSE-150 group was 29.11μmol/L and in TE-3 group of 27.47μmol/L.The proportion of G0/G1 phase was increased(both P<0.001)However,in the group treated with concentrations of MK88650μmol/L,G2/M phase was increased(P<0.05).The expression of cleaved-caspase-3 and LC-3A/B-Ⅱwere increased with MK886(both P<0.001).Conclusions MK886 inhibits proliferation of human esophageal carcinoma cell lines and induces apoptosis.