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ID:54601557
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页数:5页
时间:2020-05-03
《Livin shRNA真核表达载体构建及其对HepG2细胞化疗敏感性的影响-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、5I卷第l2期LLI东大学学报(医学版)201312J1]VO1.51NO.12JOURNALOFSHANDONGUNIVERSITY(HEALTHSCIENCES)Dec.2Ol3文章编号:1671—7554(2013)12—0020—05DOI:10.6040/j.issn.1671-7554.0.2013.011LivinshRNA真核表达载体构建及其对HepG2细胞化疗敏感性的影响徐威,常宏,翟云鹏(山东大学附属省立医院普外科,济南250021)摘要:目的构建针对Livin基因的短发夹RNA(shRNA)真核表达载体,探讨其对肝肿瘤HepG2细胞化疗敏感性的影响。
2、方法设计并构建针对Livin基因的shRNA真核表达载体pSD11一u6/Neo/GFP/Livin,脂质体法转染肝肿瘤HepG2细胞。荧光定量PCR、Westernblotting检测细胞LivinmRNA和蛋白的相对表达水平。顺铂(2.0mg/L)处理后,MTT法检测细胞生长抑制率。结果测序分析证实针对Livin基因的shRNA真核表达载体构建成功,转染肝肿瘤HepG2细胞后可降低LivinmRNA和蛋白的相对表达量(P<0.05),顺铂对转染Livin表达载体的HepG2细胞抑制率明显高于未转染细胞(P<0.05)。结论成功构建LivinshRNA真核表达载体,且有
3、效抑制HepG2细胞中Livin基因的表达,增强肝肿瘤细胞对化疗药物的敏感性。关键词:Livin基因;RNA干扰;肝细胞肝肿瘤中图分类号:R735.7文献标志码:AConstructionofLivinshRNAeukaryoticexpressionvectoranditsinfluenceonchemotherapeuticsensitivityofHepG2cellsXUWei,CHANGHong,ZHAIYun·peng(DepartmentofGeneralSurgery,ShandongProvincialHospitalAfiliatedtoShandong
4、University,Jinan250021,China)Abstract:ObjectiveToconstructashorthairpinRNA(shRNA)eukaryoticexpressionvectortargetingtotheLivingene,andexploreitsinfluenceonchemotherapeuticsensitivityofHepG2cells.MethodsTheLivinshRNAeukaryoticexpressionvectorspSDI1一U6/Neo/GFP/livinweredesigned,constructeda
5、ndtransfectedstablyintoHepG2cells.TheLivinmRNAandproteinrelativeexpressionweredetectedbyfluorescencequantitativePCRandWesternblottingMTT.wasusedtoevaluatetheinhibitionrateofHepG2cellsafterexposuretocisplatin(2.0mg/L).ResultsTheDNAse—quencingconfirmedthattheLivinshRNAeukaryoticexpressionve
6、ctorsweresuccessfullyconstructed.TransfectionofthevectorscouldsignificantlydecreasetheLivinmRNAandproteinrelativeexpressionofHepG2cells(P<0.05).TheinhibitionrateofcisplatinintransfectedcellswereobviouslyhigherthanthoseintheuntransfectedcellsfP<0.05).ConclusionTheshRNAeukaryoticexpressionv
7、ectorstargetingtotheLivingeneareconstructedsuccessfully.TransfectionofthevectorsCanefectivelydecreasetheexpressionofLivingeneinHepG2cells,andincreasethesensi—tivityofHepG2cellstochemotherapy.Keywords:Livingene;RNAinterference;HehapocellularneoplasmLivin是凋亡抑制蛋白(inhib
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