肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2014年
6期
366-372
,共7页
王涛%马梁明%牛燕燕%任瑞瑞
王濤%馬樑明%牛燕燕%任瑞瑞
왕도%마량명%우연연%임서서
肿瘤%多药耐药%三氧化二砷%谷胱甘肽
腫瘤%多藥耐藥%三氧化二砷%穀胱甘肽
종류%다약내약%삼양화이신%곡광감태
Neoplasms%Multidrug resistance%Arsenic trioxide%Glutathione
目的 研究应用丁硫氨酸亚砜胺(BSO)降低谷胱甘肽(GSH)含量后,三氧化二砷(As2O3)对K562/ADM细胞的诱导凋亡效应,及其对多药耐药相关蛋白1(MRP1)的抑制作用.方法 As2O3组(0.5、2.0、5.0 μmol/L)单独及联合100 μmol/L BSO作用于K562/ADM细胞,应用四甲基偶氮唑蓝(MTT)比色法检测K562/ADM细胞的增殖活性;Annexin V/PI标记法观察K562/ADM细胞的凋亡效应;分光光度法检测K562/ADM细胞内GSH含量变化;流式细胞术(FCM)检测的MRP1蛋白水平表达变化;反转录-聚合酶链反应(RT-PCR)方法检测MRP1的编码基因mRNA的表达变化.结果 降低GSH含量后,临床剂量As2O3(0.5、2.0 μmol/L)联合BSO 24 h内即可抑制K562/ADM细胞的增殖活性,诱导K562/ADM细胞发生凋亡,72 h单用As2O3(0.5、2.0 μmol/L)凋亡率为(8.32±2.11)％、(16.75±3.56)％,GSH含量降低后,两剂量组细胞的凋亡率分别为(82.15±9.28)％和(92.72±9.41)％.72 h后,临床剂量As2O3联合BSO抑制MRP1的荧光强度分别为8.20±0.92和10.40±2.33,明显低于单用高剂量As2O3组的荧光强度(21.30±3.09);两药联合对于MRP1 mRNA的作用效果也明显强于单用高剂量As2O3组.结论 GSH的含量变化与As2O3的作用效果密切相关,As2O3联合BSO可有效诱导K562/ADM细胞发生凋亡,可有效抑制MRP1及其编码基因mRNA的表达.
目的 研究應用丁硫氨痠亞砜胺(BSO)降低穀胱甘肽(GSH)含量後,三氧化二砷(As2O3)對K562/ADM細胞的誘導凋亡效應,及其對多藥耐藥相關蛋白1(MRP1)的抑製作用.方法 As2O3組(0.5、2.0、5.0 μmol/L)單獨及聯閤100 μmol/L BSO作用于K562/ADM細胞,應用四甲基偶氮唑藍(MTT)比色法檢測K562/ADM細胞的增殖活性;Annexin V/PI標記法觀察K562/ADM細胞的凋亡效應;分光光度法檢測K562/ADM細胞內GSH含量變化;流式細胞術(FCM)檢測的MRP1蛋白水平錶達變化;反轉錄-聚閤酶鏈反應(RT-PCR)方法檢測MRP1的編碼基因mRNA的錶達變化.結果 降低GSH含量後,臨床劑量As2O3(0.5、2.0 μmol/L)聯閤BSO 24 h內即可抑製K562/ADM細胞的增殖活性,誘導K562/ADM細胞髮生凋亡,72 h單用As2O3(0.5、2.0 μmol/L)凋亡率為(8.32±2.11)％、(16.75±3.56)％,GSH含量降低後,兩劑量組細胞的凋亡率分彆為(82.15±9.28)％和(92.72±9.41)％.72 h後,臨床劑量As2O3聯閤BSO抑製MRP1的熒光彊度分彆為8.20±0.92和10.40±2.33,明顯低于單用高劑量As2O3組的熒光彊度(21.30±3.09);兩藥聯閤對于MRP1 mRNA的作用效果也明顯彊于單用高劑量As2O3組.結論 GSH的含量變化與As2O3的作用效果密切相關,As2O3聯閤BSO可有效誘導K562/ADM細胞髮生凋亡,可有效抑製MRP1及其編碼基因mRNA的錶達.
목적 연구응용정류안산아풍알(BSO)강저곡광감태(GSH)함량후,삼양화이신(As2O3)대K562/ADM세포적유도조망효응,급기대다약내약상관단백1(MRP1)적억제작용.방법 As2O3조(0.5、2.0、5.0 μmol/L)단독급연합100 μmol/L BSO작용우K562/ADM세포,응용사갑기우담서람(MTT)비색법검측K562/ADM세포적증식활성;Annexin V/PI표기법관찰K562/ADM세포적조망효응;분광광도법검측K562/ADM세포내GSH함량변화;류식세포술(FCM)검측적MRP1단백수평표체변화;반전록-취합매련반응(RT-PCR)방법검측MRP1적편마기인mRNA적표체변화.결과 강저GSH함량후,림상제량As2O3(0.5、2.0 μmol/L)연합BSO 24 h내즉가억제K562/ADM세포적증식활성,유도K562/ADM세포발생조망,72 h단용As2O3(0.5、2.0 μmol/L)조망솔위(8.32±2.11)％、(16.75±3.56)％,GSH함량강저후,량제량조세포적조망솔분별위(82.15±9.28)％화(92.72±9.41)％.72 h후,림상제량As2O3연합BSO억제MRP1적형광강도분별위8.20±0.92화10.40±2.33,명현저우단용고제량As2O3조적형광강도(21.30±3.09);량약연합대우MRP1 mRNA적작용효과야명현강우단용고제량As2O3조.결론 GSH적함량변화여As2O3적작용효과밀절상관,As2O3연합BSO가유효유도K562/ADM세포발생조망,가유효억제MRP1급기편마기인mRNA적표체.
Objective To investigate the apoptosis-inducing effect,inhibiting multidrug resistanceassociated protein 1 (MRP-1) and mRNA expression of arsenic trioxide (As2O3) and buthionine sulfoximine (BSO) in multidrug-resistant cell K562/ADM.To compare the effect of As2O3 and the combined group.To determine the effect of intracellular glutathione (GSH) content on the arsenic effect.Methods The arsenic group (0.5 μmol/L,2.0 μmol/L,5.0 μmol/L) solo or combined BSO (100 μmol/L) applied in multidrugresistant cell K562/ADM.The cell proliferating activity was assessed with MTT assay.The cell apoptosis effect was detected by Annexin-V and propidium iodide (PI) staining.Intracellular GSH contents were measured using GSH Assay Kit by spectrophotometry.MRP1 expression was determined by flow cytometry.MRP1 mRNA expression were directed by semi-quantitative RT-PCR.Results With the GSH contents were degraded by the combination of clinic dose arsenic group (0.5 μmol/L,2 μmol/L) and BSO (100 μmol/L),the K562/ADM cell proliferating activity was obviously inhibited and the cell apoptosis-inducing effect was increased in 24 hours.In 72 hours,the rate of apoptosis with arsenic (0.5 μmol/L,2 μmol/L) were (8.32± 2.11) ％,(16.75±3.56) ％.After the GSH contents were degraded,the rates of apoptosis in the combination group (clinic dose arsenic group) were (82.15±9.28) ％ and (92.72±9.41) ％.The fluorescence intensity of MRP1 in 72 hours of the combination group (clinic dose arsenic group) was 8.20±0.92 and 10.40±2.33.The MRP1 attenuated effect of the combination group (clinic dose arsenic group) was obviously stronger than that of the high dose arsenic group (21.30±3.09).Conclusions The intracellular GSH contents closely correlate with the arsenic effect.The cell apoptosis-inducing effect of the combination of clinic dose arsenic and BSO on K562/ADM cell is obvious increased.The combination of clinic dose arsenic and BSO obviously inhibit MRP1 expression and MRP1 mRNA expression in K562/ADM cell.
