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ID:32867951
大小:2.08 MB
页数:40页
时间:2019-02-16
《培美曲塞对肺腺癌a549细胞中skp2蛋白表达的影响及意义》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、南昌大学硕士学位论文培美曲塞对肺腺癌A549细胞中SKP2蛋白表达的影响及意义姓名:万琴申请学位级别:硕士专业:肿瘤学指导教师:王顺金201205摘要目的:探讨培美曲塞(PMX)对肺腺癌A549细胞中SKP2(S期激酶相关蛋白2)蛋白表达的影响及其意义。方法:本实验根据PMX浓度设一个空白对照组和四个实验组分别为:01ttM(A组)、0.019M(B组)、0.1p.M(C组)、1I.tM(D组)、101aM(E组)。采用MTT法检测不同浓度PMX处理24h、48h、72h后各组A549细胞的增殖抑制;利用流式细胞周期技术
2、分析不同浓度PMX处理48h后各组A549细胞周期分布;通过Westernblot法检测不同浓度PMX处理48h后各组A549细胞中SKP2蛋白表达。结果:PMX能明显抑制A549细胞增殖,以109M、72h作用后细胞抑制率(78.61±3.66)%最高(尸均<0.01);与A组相比,C、D、E组S期细胞显著增多(P3、相对表达水平显著增加(尸均<0.01),且在一定的浓度范围内,随着PMX浓度增加其蛋白表达量也增加。结论:PMX能明显抑制A549细胞增殖且将其阻滞于S期,PMX处理后的A549细胞SKP2蛋白表达上调,这可能影响PMX对肺腺癌的化疗疗效。关键词:培美曲塞:A549细胞:SKP2AbstractABSTRACTobjective:Toexploretheinfluenceofs-phasel【inaseassociatedprotein2(SKP2)expressionanditssignificanceinlungad4、enocarcinomaA549cellsafterthetreatmentofPemetrexed(PMX).Methods:AccordingtotheconcentrationofPMX,wesetonecontrolgroupandfourexperimentalgroups:OlaM(groupA),0.01uM(groupB),O.1laM(groupC),1uM【groupD),10pM(groupE).ThegrowthofA549cellshandledbydifferentconcentrationP5、MXfor24h,48h,72h,wastestedbyMTTassay;TheeffectofdifferentconcentrationPMXhandlingfor48honthecellcyclephasedistributionofA549cellswasanalyzedbyflowcytometry;TheexpressionofSKP2proteinofA549cellshandledbydifferentconcentrationPMXfor48hwasdetectedbyWesternblot.Resul6、ts:MTTassayshowedthatPMXcouldsignificantlyinhibitthegrowthofA549cells,andthehighestinhibitionratioWas(78.61±3.66)%attheconcentrationofIOIaMwhenA549cellswereculturedfor72h∽7、ofs-phasecellswasincreasing、析ththeconcertrationofPMX.ThedifferenceofSKP2proteinexpressionamongeachgroupwassignificantly(P<0.01),andcomparedwithcontrolgroupSKP2proteinexpressionsignificantlyincreasedinexperimentalgroups(P<0.01).Thatincreasingwasobviouswiththeincre8、asingofPMXincertainconcentrationrange.Conclusion:PMXcouldinhibitthegrowthofA549cellsandarrestedthemins—phase.TheexpressionofSKP2proteininPMX—treatedA549cellsWa
3、相对表达水平显著增加(尸均<0.01),且在一定的浓度范围内,随着PMX浓度增加其蛋白表达量也增加。结论:PMX能明显抑制A549细胞增殖且将其阻滞于S期,PMX处理后的A549细胞SKP2蛋白表达上调,这可能影响PMX对肺腺癌的化疗疗效。关键词:培美曲塞:A549细胞:SKP2AbstractABSTRACTobjective:Toexploretheinfluenceofs-phasel【inaseassociatedprotein2(SKP2)expressionanditssignificanceinlungad
4、enocarcinomaA549cellsafterthetreatmentofPemetrexed(PMX).Methods:AccordingtotheconcentrationofPMX,wesetonecontrolgroupandfourexperimentalgroups:OlaM(groupA),0.01uM(groupB),O.1laM(groupC),1uM【groupD),10pM(groupE).ThegrowthofA549cellshandledbydifferentconcentrationP
5、MXfor24h,48h,72h,wastestedbyMTTassay;TheeffectofdifferentconcentrationPMXhandlingfor48honthecellcyclephasedistributionofA549cellswasanalyzedbyflowcytometry;TheexpressionofSKP2proteinofA549cellshandledbydifferentconcentrationPMXfor48hwasdetectedbyWesternblot.Resul
6、ts:MTTassayshowedthatPMXcouldsignificantlyinhibitthegrowthofA549cells,andthehighestinhibitionratioWas(78.61±3.66)%attheconcentrationofIOIaMwhenA549cellswereculturedfor72h∽7、ofs-phasecellswasincreasing、析ththeconcertrationofPMX.ThedifferenceofSKP2proteinexpressionamongeachgroupwassignificantly(P<0.01),andcomparedwithcontrolgroupSKP2proteinexpressionsignificantlyincreasedinexperimentalgroups(P<0.01).Thatincreasingwasobviouswiththeincre8、asingofPMXincertainconcentrationrange.Conclusion:PMXcouldinhibitthegrowthofA549cellsandarrestedthemins—phase.TheexpressionofSKP2proteininPMX—treatedA549cellsWa
7、ofs-phasecellswasincreasing、析ththeconcertrationofPMX.ThedifferenceofSKP2proteinexpressionamongeachgroupwassignificantly(P<0.01),andcomparedwithcontrolgroupSKP2proteinexpressionsignificantlyincreasedinexperimentalgroups(P<0.01).Thatincreasingwasobviouswiththeincre
8、asingofPMXincertainconcentrationrange.Conclusion:PMXcouldinhibitthegrowthofA549cellsandarrestedthemins—phase.TheexpressionofSKP2proteininPMX—treatedA549cellsWa
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