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1、哈爾濱醫(yī)科大學碩士學位論文人肺腺癌Anip973/CDDP耐藥細胞系的建立及生物學特性分析姓名:張長海申請學位級別:碩士專業(yè):腫瘤學指導教師:陳公琰20070501人肺腺癌Anip973/CDDP耐藥細胞系的建立及生物學特性分析中文摘要目的建立人肺腺癌耐藥細胞模型Anip973/CDDP并檢測其生物學特性方法本研究應用人肺腺癌細胞系Anip973采用順鉑逐步增加劑量法誘導建立人肺腺癌耐藥細胞模型Anip973/CDDP細胞計數(shù)法繪制生長曲線計算其倍增時間光學顯微鏡和掃描電鏡觀察其形態(tài)流式細
2、胞儀分析其細胞周期MTT法測定細胞IC50計算其耐藥指數(shù)結(jié)果本研究經(jīng)過24個月建立了人肺腺癌耐藥細胞系Anip973/CDDP通過Anip973/CDDP生物學特性分析發(fā)現(xiàn)(1)光鏡及電子顯微鏡觀察Anip973/CDDP細胞與Anip973細胞比較細胞形態(tài)不規(guī)則呈長偽足的多角形及體積增大胞質(zhì)內(nèi)細胞器比較發(fā)達線粒體豐富(2)與親代細胞相比Anip973/CDDP細胞生長曲線的倍增時間無明顯差異(3)細胞周期分析發(fā)現(xiàn)耐藥細胞系Anip973/CDDP的G0-G1期細胞明顯減少為52.
3、18S期細胞明顯增多為28.60%而親代細胞系Anip973的上述各周期的百分比分別為66.11%和16.37(p<0.05)兩種細胞系G2-M期細胞無明顯變化提示耐藥細胞的DNA合成增加經(jīng)MTT法測定Anip973/CDDP細胞比較親代細胞的CDDP半數(shù)致死濃度IC50增大至27.26倍該耐藥細胞系同時對伊立替康吉西他濱達卡巴嗪足葉乙甙諾維本表阿霉素藥物也產(chǎn)生了交叉耐藥耐藥倍數(shù)分別為20.3319.967.105.345.274.38而對異環(huán)磷酰胺氟尿嘧啶
4、相對敏感耐藥倍數(shù)分別為1.251.09均P0.05結(jié)論Ainp973/CDDP是可靠的人肺腺癌耐藥細胞系它的建立為進一步探討順鉑致人NSCLCMDR的發(fā)生機制奠定了基礎關(guān)鍵詞肺腺癌Anip973/CDDP細胞系順鉑多藥耐藥2哈爾濱醫(yī)科大學碩士學位論文EstablishmentofaHumanLungAdenocarcinomaDrug-resistanceCellLineAnip973/CDDPandanalysisofitsbiologicalpropertiesAbstrac
5、tObjectiveToestablishahumanlungadenocarcinomamultidrug-resistantcelllineAnip973/CDDPandtostudyitsbiologiccharacteristics.MethodsThehumanlungadenocarcinomamultidrug-resistantcellline(A973/CDDP)wasestablishedbyinducinghumanlungadenocarcinomacelllineAni
6、p973withstep-wiseincreasingconcentrationsofcisplatin,whichusedtobethefirst-linechemotherapeuticdrugforlungcacer.Thecellgrowthcurveandthedoublingtimeweredeterminedbycounting.Changesincellularmorphologyandultrastructurewereobservedunderinverted-microsc
7、opeandscanningelectronmicroscoperespectively.Thecellcyclesweredeterminedbyflowcytometry.ThechemosensitivitiesofAnip973/CDDPandAnip973tocisplatin,irinotecan,gemcitabine,dacarbazine,etoposide,navelbine,pharmorubicin,isofosfamideand5-fluorouracilweretes
8、tedandIC50measuredbyMTT.ResultsThehumanlungadenocarcinomamultidrug-resistantcelllineAnip973/CDDPwasestablishedsuccessfullyfor24months.Itsbiologiccharacteristicswere:(1)ThecellmorphousofA973/CDDPcellsweremoreirregularthanA973cells,andtherewassignifica