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《地塞米松對(duì)順鉑誘導(dǎo)人肺腺癌細(xì)胞SPC—A1凋亡的抑制作用及分子機(jī)制-論文.pdf》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、第3l卷第3期生物醫(yī)學(xué)工程學(xué)雜志V0I．3lNo．32014年6月JournalofBiomedicalEngineeringJune2014地塞米松對(duì)順鉑誘導(dǎo)人肺腺癌細(xì)胞SPC—A1凋亡的抑制作用及分子機(jī)制曹菲張智慧1(成都市西區(qū)醫(yī)院腫瘤科，成都610036)2(四川省腫瘤醫(yī)院腫瘤內(nèi)科，成都610041)摘要：研究地塞米松(DEX)對(duì)順鉑(cDDP)引起的人肺腺癌細(xì)胞SPC—A1凋亡的抑制作用及可能的分子機(jī)制。將不同濃度的DEX分別加入人肺腺癌細(xì)胞SPC—Al體外誘導(dǎo)培養(yǎng)24h后，再用不同濃度CDDP處理
2、SPC—A1細(xì)胞48h。MTT法檢測(cè)細(xì)胞存活率；RT—PCR方法檢測(cè)1tmol／IDEX誘導(dǎo)培養(yǎng)不同時(shí)間后SPC—A1細(xì)胞內(nèi)血清／糖皮質(zhì)激素一誘導(dǎo)激酶(SGK一1)和有絲分裂原蛋白激酶(MKP一1)的表達(dá)；用生物素標(biāo)記的抗糖皮質(zhì)激素受體(GR)抗體對(duì)SPC—A1細(xì)胞行免疫組化染色來(lái)檢測(cè)GR的表達(dá)。MTT測(cè)定結(jié)果顯示，SPC—A1細(xì)胞經(jīng)DEX誘導(dǎo)后，對(duì)CDDP所致的凋亡有抵抗作用并與DEX濃度呈劑量依賴關(guān)系。RT—PCR檢測(cè)到DEX誘導(dǎo)培養(yǎng)能提高SGK—l在SPC—A1細(xì)胞中的表達(dá)，表達(dá)量隨時(shí)間延長(zhǎng)而增加；但
3、未檢測(cè)到MKP一1的表達(dá)。免疫組化檢測(cè)顯示SPC—A1細(xì)胞經(jīng)DEX誘導(dǎo)后GR上調(diào)，細(xì)胞內(nèi)GR表達(dá)的陽(yáng)性細(xì)胞數(shù)明顯高于對(duì)照組。結(jié)果表明DEX對(duì)CDDP引起SPC—A1細(xì)胞的凋亡有抑制作用?？赡艿姆肿訖C(jī)制是通過(guò)上調(diào)細(xì)胞內(nèi)GR表達(dá)，進(jìn)而上調(diào)其通路下游抗凋亡蛋白SGK一1的表達(dá)，導(dǎo)致SPC—Al細(xì)胞產(chǎn)生抗凋亡作用。關(guān)鍵詞：地塞米松；糖皮質(zhì)激素受體；肺癌；化療；抗凋亡中圖分類號(hào)R73—36文獻(xiàn)標(biāo)志碼ADOI：10．7507／1OO卜55l5．20140122TheInhibitoryEffectsofDexameth
4、asoneonCisplatinInducedApoptosisofHumanLungAdenocarcinomaCellSPC—A1andItsMolecularMechanismCAoFeiZHANGZhihui1(Departmento_，Ontology，ChengduWesternHospital，Chengdu610036，China)2(Department0，MedicalOmology，SichuanProvincialTumorHospital，Chengdu610041，China)A
5、bstract：Theaimofthisstudyistoinvestigatetheapoptoticinhibitionanditsmolecularmechanismofdexametha—sone(DEX)actingoncisplatin(CDDP)一inducedapoptosisofhumanlungadenocarcinomacellSPC—A1．SPC—A1ceilswerepre—culturedinvitrofor24hourswithDEXindifferentconcentrati
6、onsandthenCDDPwasaddedindifferentconcentrationsforculturingforfurther48hours。ThesurvivalratesofthecellsweredeterminedbyMTT．Theex—pressionofserum／glucocorticoid—inducedkinase(SGK一1)andmitogen—activatedproteinkinasephosphatase一1(MKP一1)inSPC—A1ceilsa{terbeing
7、culturedby1~mol／IDEXatdifferenttimewasdetectedbysemi—quantitativeRT～PCRtechnology．Theexpressionofglucocorticoidreceptor(GR)inSPC—A1cellswasmeasuredbyimmunohisto—chemistry(IHC)withbiotin—labeledanti—GR．TheresultsofMTTshowedthatSPC—AlcellshadresistancetoCDDP
8、inducedapoptosiswithpre—culturedDEXandtheresistanceintensitypresentedDEXconcentration-dependent．TheexpressingquantityofSGK一1inSPC—A1cellsstimulatedbyDEXcouldbeelevatedandincreasedwithintentionoftime．buttheexp
