VEGF 反義寡核苷酸抑制膽囊癌生長增殖和凋亡的體外實(shí)驗(yàn)研究.doc

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1、VEGF反義寡核苷酸抑制膽囊癌生長增殖和凋亡的體外實(shí)驗(yàn)研究作者：龐作良,李海軍,毛拉艾沙·買買提【摘要】目的體外研究Oligofectamine介導(dǎo)的VEGF反義寡核苷酸轉(zhuǎn)染對膽囊癌GBCSD細(xì)胞生長、增殖和凋亡的影響。方法運(yùn)用Oligofectamine介導(dǎo)VEGF反義寡核苷酸(Antisenseoligodeoxynucleotides，ASODN)和錯義寡核苷酸(ScrambledOligodeoxynucleotide，SODN)轉(zhuǎn)染人膽囊癌GBCSD細(xì)胞，MTT法測定轉(zhuǎn)染后各組細(xì)胞的生長曲線及抑制率，流式細(xì)胞術(shù)檢測轉(zhuǎn)染后不同

2、時(shí)間各組細(xì)胞凋亡情況。結(jié)果SODN、SODN+Oligofectamine對GBCSD細(xì)胞生長無顯著影響(P＞0.05)，而ASODN及ASODN+Oligofectamine則能顯著抑制細(xì)胞生長(P<0.05)，且ASODN+Oligofectamine對細(xì)胞生長增殖抑制作用較ASODN組更為強(qiáng)(P<0.05)。流式細(xì)胞術(shù)檢測結(jié)果發(fā)現(xiàn)ASODN+Oligofectamine能促進(jìn)GBCSD細(xì)胞凋亡(P<0.05)。結(jié)論VEGFASODN轉(zhuǎn)染能抑制膽囊癌GBCSD細(xì)胞生長和增殖，Oligofectamine介導(dǎo)能明

3、顯增強(qiáng)VEGFASODN的抑制作用；Oligofectamine介導(dǎo)VEGF反義寡核苷酸轉(zhuǎn)染能促進(jìn)膽囊癌GBCSD細(xì)胞的凋亡?！娟P(guān)鍵詞】膽囊癌細(xì)胞株GBCSD；VEGF；反義寡核苷酸；Oligifectamine；凋亡　　ExperimentofOligofectamineMediatedVEGFASODNTransfectiononGBCSDCellGrowth,ProliferationandApoptosisinVitro　　Abstract：ObjectiveToinvestigatetheeffectofoligofect

4、aminemediatedVEGFASODNtransfectiononGBCSDcellgrowth,proliferationandapoptosisinvitro.MethodsGBCSDcellsweretransfectedwithVEGFASODNandSODNmediatedbyoligofectamine.ThechangeoftransfectedGBCSDcellsgrowthandproliferationactivitybyMTT,andapoptosisbyflowcytometrywereassessed

5、.ResultsThetransfectedGBCSDcellsgrowthandproliferationactivitytestedingroupsofSODNandSODN+oligofectaminehadnodifferenceascomparedwithcontrolgroup(P>0.05),butasforthecellsofASODNandASODN+oligofectamine,thegrowthandproliferationactivityweresignificantlyinhibited(P<0.

6、05),moreover,theeffectofASODN+oligofectaminewasmoresignificantthanthatofASODN(P<0.05).TheresultsofflowcytometryrevealedASODN+oligofectaminecouldacceleratetransfectedGBCSDcellsapoptosis(P<0.05).ConclusionVEGFASODNcouldinhibitGBCSDcellsgrowth,proliferationandacceler

7、ateapoptosis.TheoligofectaminecouldstrengthentheeffectofASODN.6　　Keywords：GBCSDcell;VEGF;ASODN;Oligifectamine;Apoptosis　　0引言　　VEGF通過與其特異性受體(VEGFR)結(jié)合，刺激血管內(nèi)皮細(xì)胞增殖和遷移，促進(jìn)新生血管生成，在腫瘤的生長和轉(zhuǎn)移中起重要作用。抑制VEGFR信號轉(zhuǎn)導(dǎo)通路的任何一個(gè)環(huán)節(jié)，就可能防止腫瘤的進(jìn)展與轉(zhuǎn)移。ASODN因有專一序列的寡聚脫氧核苷酸，故能特異阻斷靶基因的復(fù)制、轉(zhuǎn)錄和翻譯，是目前迅速發(fā)展的一

8、種新型的基因治療藥物[1]。Oligofectamine是一種專門用于寡核苷酸轉(zhuǎn)染的陽離子脂質(zhì)體，具有較高轉(zhuǎn)染率。本實(shí)驗(yàn)體外運(yùn)用Oligofectamine介導(dǎo)的VEGFASODN轉(zhuǎn)染人膽囊癌