資源描述：
《shrna干擾沉默mcl-1基因?qū)α馨土黾毎翟鲋车挠绊憽酚蓵T上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在工程資料-天天文庫。
1、shRNA干擾沉默Mcl-1基因?qū)α馨土黾毎翟鲋车挠绊懺拦庑?，張明智1，張旭東1，許偉朋2，陳新峰1，陳昱丞1，曹玲1，楊黎3，盛譽喬3（450052鄭州，鄭州大學第一附屬醫(yī)院：腫瘤科1，骨科2，河南省高等學校臨床醫(yī)學重點學科開放實驗室3）［摘要］目的研究慢病毒介導的髓細胞白血病基因（Mcl-1）短發(fā)夾狀RNA（shRNA）干擾對淋巴瘤細胞系SNK-6增殖和凋亡的影響。方法設(shè)計以Mcl-1為靶點的短發(fā)夾狀RNA（shRNA）并構(gòu)建攜帶此shRNA的慢病毒載體，轉(zhuǎn)染淋巴瘤細胞系SNK-6，熒光定量PCR(qPCR)及Westernblot
2、方法檢測病毒感染前后Mcl-1mRNA及蛋白表達變化，四甲基偶氮唑鹽法（MTT法）和流式細胞儀分析細胞增殖和凋亡變化的情況。結(jié)果成功構(gòu)建Mcl-1-shRNA慢病毒表達載體，并可有效感染淋巴瘤細胞株SNK-6，感染后SNK-6細胞Mcl-1基因的mRNA水平及蛋白水平表達明顯下調(diào),，MTT法檢測顯示慢病毒介導的Mcl-1基因shRNA干擾與對照病毒組相比可明顯抑制SNK-6細胞增殖[（31.6±3.3）%vs（5.8±2.7）%，P<0.01]，流式細胞儀測定感染后細胞凋亡明顯高于對照病毒組[（28.9±2.1）%vs(5.3±1.5)%,
3、P<0.01]。結(jié)論慢病毒介導的Mcl-1基因shRNA干擾技術(shù)可特異性阻斷SNK-6細胞Mcl-1基因的表達，抑制細胞的增殖并促進細胞的凋亡。［關(guān)鍵詞］淋巴瘤；髓細胞白血病基因-1；短發(fā)夾RNA；凋亡；慢病毒載體[中圖法分類號][文獻標志碼]ATheeffectsofshRNAinterferencetargetingMcl-1onproliferationoflymphomacelllineYueGuangxing1，ZhangMingzhi1，ZhangXudong1，XuWeipeng2，ChenXinfeng1，ChenYuche
4、ng1，CaoLing1，YangLi3，ShengYuqiao3(Departmentof1oncology,2orthopedics,3InstituteofClinicalMedicine,theFirstAffiliatedHospitalofZhengzhouUniversity，ZhengzhouHenan450052,China)［Abstract］ObjectiveToexploretheeffectsoflentivirus-mediatedshRNAinterference(RNAi)targetingtheMyeloi
5、dleukemia-1（Mcl-1）onproliferationandapoptosisoflymphomacelllineSNK-6.MethodsShorthairpinRNA(shRNA)targetingMcl-1wasdesignedandthelentiviralvectorcarryingtheshRNAwasconducted.AfterinfecttionoflentiviralvectorinSNK-6cells,RealtimePCRandwesternblotwereusedtoanalyzetheexpressi
6、onofMcl-1mRNAandprotein.MTTassayandflowcytometrywereemployedtodetectthestatusofproliferationandapoptosis.ResultsTheshRNAtargetingMcl-1wassuccessfullyinsertedintothelentiviralvector,whichcaneffectivelyinfectthelymphomacelllineSNK-6.AfterinfectingtheSNK-6cells,theexpressiono
7、fMcl-1mRNAandproteinwasobviouslydecreasedandMTTassaysuggestedthattheproliferationofSNK-6cellswasinhabitedbyshRNAinterferencetargetingMcl-1againstcontrol-shRNAgroup(31.6%±3.3%vs5.8%±2.7%,P<0.001).FlowcytometryshowedthattheapoptosisrateofMcl-1-shRNA-infectedcellswassignifica
8、ntlyhigherthanthecontrolgroup(28.9%±2.1%vs5.3%±1.5%,P<0.001).ConclusionLentivirus-mediate
