维生素E琥珀酸酯联合5-氟尿嘧啶和5′-脱氧氟脲苷促进人结肠癌细胞凋亡
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摘要
目的探讨维生素E琥珀酸酯(VES)联合5-氟尿嘧啶(5-FU)和5′-脱氧氟脲苷(5′-DFUR)对人结肠癌细胞株LS174T凋亡的影响。方法 LS174T细胞常规培养,分6组(0,20μmol/L VES,1μmol/L 5-FU,2μmol/L 5′-DFUR,20μmol/L VES+1μmol/L 5-FU,20μmol/L VES+2μmol/L 5′-DFUR),处理24 h,倒置显微镜观察细胞形态,MTS法检测细胞的增殖情况;Western blot法检测细胞bid、bax、bcl-2蛋白的表达情况;流式细胞仪技术检测细胞凋亡情况。结果 VES分别与5-FU、5′-DFUR联合能够抑制人结肠癌细胞的增殖,增强bid、bax蛋白的表达,降低bcl-2蛋白的表达,促进细胞发生凋亡。结论 VES分别与5-FU、5′-DFUR联合作用时能抑制细胞增殖,诱导细胞凋亡,其中VES+5′-DFUR联合作用效果最为显著。这可能与细胞中促凋亡蛋白bid、bax表达的增加、抗凋亡蛋白bcl-2表达的降低有关。
Objective To study the effect of vitamin E succinate(VES)combined with 5-fluorouracil(5-FU)and 5′-Deoxy-5-fluorouridine(5′-DFUR)on the apoptosis of human colon cancer cell line LS174 T in vitro. Methods LS174 T cells were routinely cultured in 6 groups(0,20 μmol/L VES,1 μmol/L 5-FU,2 μmol/L5′-DFUR,20 μmol/L VES + 1 μmol/L 5-FU,20 μmol/L VES + 2 μmol/L 5′-DFUR)for 24 h. The morphology ofcell was observed by inverted microscope,cell proliferation was detected by MTS;expression of bid,Bax,bcl-2 protein were detected by Western blot;flow cytometry technique to detect apoptosis. Results VES combined with5-FU,5′-DFUR inhibited the proliferation of human colon cancer cells,enhanced the expression of bid,Baxprotein,reduced the expression of Bcl-2 protein and promoted apoptosis. Conclusion VES combined with 5-FU,5′-DFUR can inhibit cell proliferation and induce apoptosis,and the effect of VES+5 ′-DFUR is the most signifi-cant. This may be related to the increase of the expression of Pro apoptotic protein bid,Bax and the expression ofanti apoptotic protein Bcl-2 in cells.
Objective To study the effect of vitamin E succinate(VES)combined with 5-fluorouracil(5-FU)and 5′-Deoxy-5-fluorouridine(5′-DFUR)on the apoptosis of human colon cancer cell line LS174 T in vitro. Methods LS174 T cells were routinely cultured in 6 groups(0,20 μmol/L VES,1 μmol/L 5-FU,2 μmol/L5′-DFUR,20 μmol/L VES + 1 μmol/L 5-FU,20 μmol/L VES + 2 μmol/L 5′-DFUR)for 24 h. The morphology ofcell was observed by inverted microscope,cell proliferation was detected by MTS;expression of bid,Bax,bcl-2 protein were detected by Western blot;flow cytometry technique to detect apoptosis. Results VES combined with5-FU,5′-DFUR inhibited the proliferation of human colon cancer cells,enhanced the expression of bid,Baxprotein,reduced the expression of Bcl-2 protein and promoted apoptosis. Conclusion VES combined with 5-FU,5′-DFUR can inhibit cell proliferation and induce apoptosis,and the effect of VES+5 ′-DFUR is the most signifi-cant. This may be related to the increase of the expression of Pro apoptotic protein bid,Bax and the expression ofanti apoptotic protein Bcl-2 in cells.
引文
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[2]徐本玲,周进学,袁龙,等.AKT抑制剂对结直肠癌肝转移患者肿瘤浸润T淋巴细胞生物学活性的影响[J].实用医学杂志,2018,34(3):352-356.
[3]HUANG X,ZHANG Z,JIA L,et al.Endoplasmic reticulumstress contributes to vitamin E succinate-induced apoptosis inhuman gastric cancer SGC-7901 cells[J].Cancer Lett,2010,296(1):123-131.
[4]ELAMIN YY,RAFEE S,OSMAN N,et al.Thymidine phos-phorylase in cancer;enemy or friend[J]?Cancer Microenviron,2016,9(1):33-43.
[5]张继民,刘明姬,溝井賢幸,等.单核-巨噬细胞表达的胸苷磷酸化酶增强5′-脱氧氟尿苷抗大肠癌细胞活性[J].中华医学杂志,2004,84(9):806-812.
[6]HOU L,ZHANG H,XU P,et al.Effect of vitamin E succinateon the expression of the tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)receptor in gastric cancer cells andCD4(+)T cells[J].Mol Biosyst,2015,11(11):3119-3128.
[7]SONG Y,CAI H,YIN T,et al.Paclitaxel-loaded redox-sensi-tive nanoparticles based on hyaluronic acid-vitamin E succinateconjugates for improved lung cancer treatment[J].Int J Nano-medicine,2018,13:1585-1600.
[8]YAN B,STANTIC M,ZOBALOVA R,et al.Mitochondriallytargeted vitamin E succinate efficiently kills breast tumour-initi-ating cells in a complex II-dependent manner[J].BMC Cancer,2015,15:401.
[9]YANG P,ZHAO J,HOU L,et al.Vitamin E succinate induc-es apoptosis via the PI3K/AKT signaling pathways in EC109esophageal cancer cells[J].Mol Med Rep,2016,14(2):1531-1537.
[10]许伟,范梦雪,孟婷婷,等.脱氧雪腐镰刀菌烯醇对PC12细胞凋亡的影响[J].南京农业大学学报,2016,39(5):838-844.
[11]MARSHALL K A,DANIEL S E,CAIRNS N,et al.Upregula-fion of anti-apoptotic protein Bcl-2 may be an early event in neu-todegeneration[J].Biochem Biophys Res Commun,1997,240(1):84-87.
[12]ZHANG L M,ZHAO X C,SUN W B,et al.Sevoflurane post-conditioning protects primary rat cortical neurons against oxygen-glucose deprivation/resuscitation via down-regulation in mito-chondrial apoptosis axis of Bid,Bim,Puma-Bax and Bak mediat-ed by Erk1/2[J].J Neurol Sci,2015,357(1-2):80-87.
[13]WEI M C,LINDSTEN T,MOOTHA V K,et al.BID,a mem-brane-targeted death ligand,oligomerizes BAK to release cyto-chrome C[J].Genes Dev,2000,14(16):2060-2071.