資源描述：
《人肝癌細(xì)胞總rna電轉(zhuǎn)染樹(shù)突狀細(xì)胞的體外腫瘤殺傷實(shí)驗(yàn)》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。
1、人肝癌細(xì)胞總RNA電轉(zhuǎn)染樹(shù)突狀細(xì)胞的體外腫瘤殺傷實(shí)驗(yàn)【摘要】目的：探討人肝癌細(xì)胞總RNA電轉(zhuǎn)染人樹(shù)突狀細(xì)胞(DendriticCells,DCs)后誘導(dǎo)特異性效應(yīng)T細(xì)胞及其效應(yīng)T細(xì)胞抗肝癌的特異性。方法：通過(guò)密度梯度離心法分離人外周血單核細(xì)胞，采用細(xì)胞因子體外培養(yǎng)誘導(dǎo)其成為未成熟樹(shù)突狀細(xì)胞（ImmatureDendriticCell,imDC）和成熟樹(shù)突狀細(xì)胞（MatureDendriticCell,mDC）。培養(yǎng)肝癌細(xì)胞，Trizol一步法提取人肝癌細(xì)胞總RNA。通過(guò)電轉(zhuǎn)染法將人肝癌細(xì)胞總RNA導(dǎo)入imDC內(nèi)；
2、觀測(cè)電轉(zhuǎn)染的轉(zhuǎn)染率；采用免疫細(xì)胞化學(xué)檢測(cè)電轉(zhuǎn)染前后的imDC形態(tài)及其表面標(biāo)志；通過(guò)混合淋巴細(xì)胞實(shí)驗(yàn)，獲取mDC激活的特異性效應(yīng)T細(xì)胞。用MTT法測(cè)定效應(yīng)T細(xì)胞的抗肝癌特異性。結(jié)果：電轉(zhuǎn)染方法可將人肝癌細(xì)胞總RNA導(dǎo)入imDC。電轉(zhuǎn)染前后imDC分子表達(dá)無(wú)顯著差異，mDC的分子表達(dá)與imDC相比發(fā)生了變化。轉(zhuǎn)染了人肝癌細(xì)胞總RNA的mDC可特異的激活T細(xì)胞為效應(yīng)T細(xì)胞。效應(yīng)T細(xì)胞可特異性的殺傷人肝癌細(xì)胞，而對(duì)不相關(guān)的MG-63細(xì)胞無(wú)特異性的殺傷作用。結(jié)論：電轉(zhuǎn)染不影響imDC的形態(tài)和原有的蛋白表達(dá)；轉(zhuǎn)染了人肝癌細(xì)胞
3、總RNA的mDC可特異的激活T細(xì)胞為效應(yīng)T細(xì)胞；效應(yīng)T細(xì)胞具有抗人肝癌細(xì)胞的特異性?！娟P(guān)鍵詞】樹(shù)突狀細(xì)胞肝癌細(xì)胞總RNA轉(zhuǎn)染人AbstractObjective:Toinvestiagewhetherthe13electransfectionofhumandendriticcells(DCs)withtotalRNAofhepatocarcinomacellscouldinducethespecificeffectiveTcells,thenanalysisthespecificythateffectiveTce
4、llskilltargethepatocarcinomacells.Methods:MonocytesobtainedformhumanperipheralbloodbydensityguadientcentrifugationareinducedintoimmatureDCsandmatureDCsinthepresenceofcytokinesinvitro.ThetotalRNAofhepatocarcinomacellsareextractedbyTrizolandareeletransfectedint
5、omonocyte-derivedimmatureDCs.Thesuccessrateofelectransfectionismeasured.WeuseimmunocellchemicalstainingtoobservethechangesofimDCs’morphologyandmoleculeexpressionbetweenbeforeandafterelectransfectionofhumandendriticcells(DCs)withtotalRNAofhepatocarcinomacells;
6、weharvesteffectiveTcellsbymixedlymphologyreaction,inwhichgraduallymatureDCsactivateTcells,thenweexaminethespecificitythateffectiveTcellskilltargethepatocarcinomacells.Results:ElectransfectioncanmakeRNAintoimDCsuccessfully.ThereisnosignificantdifferenceinimDCm
7、oleculeexpressionaftertheelectransfection.ThereisdifferentmoleculeexpressionbetweenimDCandmDC.DCswithtotalRNAofhepatocarcinomacellscanactivatespecificallyTcellsintoeffectiveTcellswhichcankillhepatocarcinoma13cellsspecificallywhilenotMG-63cells.Conclusion:Elet
8、ransfectiondoesn’tchangethemorphologyandmoleculeexpression;DCswithtotalRNAofhepatocarcinomacellscanmakeactivationofTcellsandspecificitythattheeffectiveTcellskillhepatocarcinomacellsisexam
