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1、臨床檢驗(yàn)雜志2013年12月第31卷第12期ChinJClinLabSci,Dec.2013,Vo1.31,No.12·9l5··基礎(chǔ)實(shí)驗(yàn)診斷研究·文章編號(hào):1001-764X(2013)12-915-04CIK增強(qiáng)順鉑對(duì)肺腺癌細(xì)胞系A(chǔ)549的殺傷作用李丹,李種,張克新,王智剛,徐斌,鄧海峰,陸明洋,李敏,周怡,孫青,劉檢,鄭曉,蔣敬庭(蘇州大學(xué)附屬第三醫(yī)院a.呼吸內(nèi)科,b.腫瘤生物診療中心,江蘇常州213000)摘要:目的研究細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞(CIK)聯(lián)合順鉑對(duì)人肺腺癌細(xì)胞系A(chǔ)549的殺傷作用。方法在體外將健康者外周血單個(gè)核細(xì)胞(PBMC)用細(xì)胞因子誘導(dǎo)形成具有
2、殺傷活性的CIK細(xì)胞。CIK、順鉑及CIK聯(lián)合順鉑分別作用A549細(xì)胞24、48h,MT/"法檢測(cè)細(xì)胞吸光度(A)值并計(jì)算各組細(xì)胞殺傷率。完全培養(yǎng)液培養(yǎng)CIK48h后,取上清液,按1:l稀釋后與A549細(xì)胞共培養(yǎng)24h,用流式細(xì)胞儀檢測(cè)A549細(xì)胞凋亡率。結(jié)果CIK、順鉑對(duì)A549的殺傷活性呈時(shí)間、劑量依賴性(P<0.05)。5g/mL順鉑聯(lián)合效靶比為10:1、20:1的CIK共同作用A549細(xì)胞24h,A549的殺傷率(%)分別為53.80±6.81,61.17±3.82,與單用順鉑及CIK相比,其對(duì)A549的殺傷率顯著升高(P<0.05)。CIK上清液可誘導(dǎo)A549
3、細(xì)胞凋亡。結(jié)論CIK可增強(qiáng)順鉑對(duì)A549細(xì)胞系的殺傷作用。關(guān)鍵詞:肺腫瘤;肺腺癌細(xì)胞株;細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞;順鉑中圖分類號(hào):R734.2文獻(xiàn)標(biāo)志碼:ACIKmayenhancethelethalefectofcisplatinonthelungadenocarcinomacelllineA549LIDan。,LIChong“,ZHANGlie—xin,WANGZhi—gang。,XUBin,DENGHai-feng,LUMing—yn,LIMin,ZHOU,SUNQ,L,uJian,ZHENGXiao,JIANGJing—ting(a.DepartmentofRes
4、piratoryMedicine,b.TumorBiologicalDiagnos~andTreat—mentCenter,theThirdAffiliatedHospitalofSoochoowUniversity,Changzhou213000,Jiangsu,China)Abstract:ObjectiveToinvestigatethelethaleffectofcytokine—inducedkillercells(CIK)combinedwithcisplatinonthehumanlungadenocarcinomacelllineA549.Methods
5、Theperipheralbloodmononuclearcells(PBMC)fromhealthypersonswereinducedtobetheCIKcellswitheytotoxicitybycytokinesinvitro.Then,A549cellswereincubatedwithCIK,cisplatinandCIKcombinedwithcisplatinfor24and48hours,respectively.ThekillratiosofA549ceilsweredetectedbytheMTTassay.AfterCIKcellswerein
6、cuba—tedwithcompleteculturemediumfor48h,thesupernatantwascollectedanddiluted1:1withmedium,andthenincubatedwithA549cellsfor24h.Next,theapoptosisrateofA549cellswasdetectedbyflowcytometry.ResultsBothCIKandcisplatinkilledA549cellsinadose—andtime—dependentmanner(P<0.05).AfterA549cellswereincu
7、batedwith5mLofcisplatincombinedwithCIKwith10:1and20:1ofeffector—targetratiosfor24h,thekillratiosofA549cellswere(53.80±6.81)%and(61.17±3.82)%,respectively,whichweresignificantlyhigherthanthatofcisplatinorCIKalone(P<0.05).ThesupernatantofCIKcellscouldinducetheapoptosisofA54