LMP1通过Survivin抑制^(60)Co诱导鼻咽癌细胞凋亡

Latent membrane protein 1 inhibits apoptosis induced by ^(60)Co irradiation via survivin triggering signal-pathway

目的 研究EB病毒编码的潜伏膜蛋白 1(LMP1)介导凋亡抑制蛋白 (Survivin)表达对辐射效应的影响。方法 利用LMP1可调控表达鼻咽癌细胞系 (Tet on LMP1HNE2 )诱导LMP1表达 ,同时 60 Co照射 5Gy ,采用形态学观察、流式细胞术和半胱氨酸蛋白酶 3(Caspase 3)活性检测等方法 ,分析LMP1表达对60 Co诱导鼻咽癌细胞凋亡的影响。用反义Survivin寡核苷酸阻断Survivin表达 ,观察Sur vivin表达阻断对60 Co辐射效应的影响。结果 LMP1表达抑制60 Co诱导鼻咽癌细胞凋亡 ,LMP1表达60 Co照射组形态学和流式细胞术检测凋亡率 (32 .7%± 2 .1% ,6 .3% )明显低于LMP1表达阴性组 (6 6 .0 %± 3.0 % ,2 9.6 % ) (P <0 .0 5 ) ;转染Survivin反义寡核酸后细胞凋亡率 (5 9.3%± 3.2 % ,3.0 % )明显高于对照组 (2 6 .0 %± 2 .6 % ,8.6 % ) (P <0 .0 5 ) ;同样转染Survivin反义寡核酸组Caspase 3活性 (3.78nmol/ 10 6)高于对照组 (2 .79nmol/ 10 6)。结论 提示LMP1通过介导Survivin表达而抑制60 Co照射诱导凋亡 ,Survivin反义核酸协同辐射诱导肿瘤细胞凋亡 ,Survivin作为增敏放射治疗靶 ,具有潜在的临床意义。

Objective Toinvestigate the anti apoptosismechanismofEBvirusencodenlatentmembrane protein1(LMP1)via the survivinsignaltransductionpathway after irradiationinduction .Methods Tet on LMP1HNE2cells ,asamodel,weredetected withmorphologicalassay ,flowcytometryandCaspase 3assayafter 60 CoirradiationwithLMP1inducedbydoxycycline.Theapoptosisintheanti sensesurvivintransfectedcellswas tested .Results Theresultsshowedthat,withLMP1expression ,theapoptosisratesfrommorphologicalassay andflowcytometrywere 32 .7%± 2 .1%and 6 .3% ,whichshowedthattheywerealllowerthanthatwithout LMP1expression (6 6 .0 %± 3.0 %and 2 9.6 % ) .Whenanti senseofsurvivinwasinduced ,theapoptosisrates were 5 9.0 %± 3.2 %and 3.0 %respectively ,andcaspase 3activitywas 3.78nmol/10 6cells,whichwerehigher thanthatofthecontrol (2 6 .0 %± 2 .6 % ,8.6 %and 2 .79nmol/10 6) .Survivinrestrainedthecellapoptosisin ducedbyirradiation ,butanti sense of survivin couldreleasethisinhibitionofcellapoptosistrigg eredbyLMP1ex pression.Conclusion LMP1inhibitstheirradiation inducedcella poptosisviatriggeringsurvivinexpression . Survivinmaybetargetedinsomecertaintherapy .