《過(guò)表達(dá)轉(zhuǎn)錄因子21抑制SMMC-7721肝癌細(xì)胞的增殖和遷移并促進(jìn)其凋亡》.pdf

《《過(guò)表達(dá)轉(zhuǎn)錄因子21抑制SMMC-7721肝癌細(xì)胞的增殖和遷移并促進(jìn)其凋亡》.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、884細(xì)胞與分子免疫學(xué)雜志(ChinJCellMolImmuno1)2015，31(7·論著·文章編號(hào)：1007—8738(2015)07—0884—05過(guò)表達(dá)轉(zhuǎn)錄因子21抑制SMMC-7721肝癌細(xì)胞的增殖和遷移并促進(jìn)其凋亡譚婧宇，張國(guó)慶，劉銳，周密，李中堂，吳忠均(重慶醫(yī)科大學(xué)附屬第一醫(yī)院肝膽外科，重慶400016)[摘要]目的探討轉(zhuǎn)錄因子21(TCF21)基因?qū)MMC-7721肝癌細(xì)胞增殖、遷移和凋亡的影響及機(jī)制。方法采用脂質(zhì)體將pcDNA3．1(+)TCF21及pcDNA3．1(+)質(zhì)

2、粒穩(wěn)定轉(zhuǎn)染入SMMC-7721細(xì)胞，分別為TCF21過(guò)表達(dá)組、空載體組、未處理組。采用反轉(zhuǎn)錄PCR檢測(cè)轉(zhuǎn)染前后TCF21mRNA水平、Westernblot法檢測(cè)TCF21蛋白水平；MTT法檢測(cè)細(xì)胞的增殖能力，劃痕實(shí)驗(yàn)檢測(cè)轉(zhuǎn)染后細(xì)胞的遷移能力；異硫氰酸熒光素標(biāo)記的膜聯(lián)素V／碘化丙啶(annexinV—FITC／PI)雙標(biāo)記結(jié)合流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡率，Westernblot法檢測(cè)轉(zhuǎn)染后KISSI、P53和基質(zhì)金屬蛋白酶9(MMP-9)的蛋白水平。結(jié)果TCF21基因轉(zhuǎn)染SMMC-7721細(xì)胞后，細(xì)

3、胞內(nèi)TCF21mRNA和蛋白水平均明顯增加。與對(duì)照細(xì)胞相比，過(guò)表達(dá)TCF21的肝癌細(xì)胞增殖能力減弱、癌細(xì)胞遷移能力降低、細(xì)胞凋亡增加。Westernblot結(jié)果顯示上調(diào)TCF21表達(dá)，可增加KISSI、P53水平并降低MMP-9的水平。結(jié)論過(guò)表達(dá)TCF21可抑制SMMC-7721肝癌細(xì)胞的增殖、遷移并促進(jìn)其凋亡。[關(guān)鍵詞]轉(zhuǎn)錄因子21(TCF21)；肝細(xì)胞肝癌；增殖；遷移；凋亡[中圖分類號(hào)]R735．7，Q279，R392-33[文獻(xiàn)標(biāo)志碼]AOver-expressionoftranscrip

4、tionfactor21inhibitstheproliferationandmigrationandpromotesapoptosisofSMMC-7721ceBsTANJingyu，ZHANGGuoqing，LIURui，ZHOUMi，LIZhongtang，WUZhongjunDepartmentofHepatobiliarySurgery，F(xiàn)irstAfiliatedHospital，ChongqingMedicalUniversity，Chongqing400016，ChinalAbs

5、tract]ObiectiveToexploretheimpactoftranscriptionfactor21(TCF21)geneonproliferation，migrationandapoptosisinSMMC-T／21hepatocelularcarcinomacellineandtherelatedmechanism．MethodsUsingLipofectamine~2000we，stablytransfectedpcDNA3．1(+)TCF21andpcDNA3．1(+)pla

6、smidsintoSMMC-7721cellsofTCF21over-expressiongroupandemptyvectorgroup，respectively．Theuntreatedcellsweresetasblankcontrolgroup．TheexpressionofTCF21mRNAandproteinwereinvestigatedbyreversetranscriptionPCRandWesternblotting．Celproliferationabilitywasdet

7、ectedbyM1_rassay．Woundhealingassaywasusedtoobservecellmigrationability．AnnexinV-FITC／Pldoublelabelingcombinedwithflowcytometrywasappliedtodeterminecellapoptosisrate．WesternblotingwasperformedtoexaminetheexpressionsofKISS1。P53andmatrixmetalloproteinas

8、e-9(MMP-9)．ResultsTCF21wasover-expressedinTCF'21-transfectedSMMC-7721cells．Comparedwiththecontrolgroups，theTCF2Iover-expressiongroupshowedrelativelyweakenedproliferationabilityandsignificantlyinhibitedmigrationabilityaswellastheincreasedapoptosis．Wes