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1、不同孕期小鼠羊水對(duì)小鼠肝癌細(xì)胞H22增殖和凋亡的影響作者：丁雅明,方廖瓊,周蘭,張弘,白晉單位：(重慶醫(yī)科大學(xué)生物醫(yī)學(xué)工程系,重慶醫(yī)科大學(xué)醫(yī)學(xué)超聲工程研究所,重慶400016) 摘 要:目的 觀察不同孕期小鼠羊水對(duì)小鼠肝癌細(xì)胞H22增殖和凋亡的影響。方法 用孕11、13、15d和17d的KM小鼠羊水-RPMI1640培養(yǎng)基培養(yǎng)H22細(xì)胞(培養(yǎng)液中羊水終濃度為10%),RPMI1640培養(yǎng)基培養(yǎng)的H22細(xì)胞作為對(duì)照組,采用MTT法檢測(cè)細(xì)胞的增殖能力;AnnexinV-FITC法檢測(cè)細(xì)胞凋亡率,透射電鏡觀察細(xì)胞形態(tài);S-P免疫熒光染色法經(jīng)激光共聚焦顯微鏡分析H22細(xì)胞中增殖細(xì)胞核抗原(PC
2、NA)和p53的表達(dá)。結(jié)果 羊水能明顯抑制H22細(xì)胞的體外增殖和PCNA的表達(dá),隨孕期的延長(zhǎng)而增高(P<0•05);并能促進(jìn)p53的表達(dá)以及誘導(dǎo)H22細(xì)胞凋亡,隨孕期的延長(zhǎng)而降低(P<0•05)。同時(shí),PCNA和p53的表達(dá)強(qiáng)度具有明顯的負(fù)相關(guān)性(r=-0.808,P<0•01)。結(jié)論 孕11、13、15、17d的小鼠羊水能抑制H22細(xì)胞增殖,誘導(dǎo)其凋亡,可能與PCNA和p53的表達(dá)有關(guān),并且孕期越早、干預(yù)時(shí)間越長(zhǎng),效果越顯著。 關(guān)鍵詞:H22細(xì)胞;羊水;凋亡;增殖細(xì)胞核抗原;p53 中圖法分類號(hào):R73-354;R73-36;R735
3、.7文獻(xiàn)標(biāo)識(shí)碼:AEffectsofmouseamnioticfluidcollectedondifferentgestationaldaysonprolifera-tionandapoptosisofH22cellsinvitroDINGYa-ming,FANGLiao-qiong,ZHOULan,ZHANGHong,BAIJin,WANGZhi-iaoDepartmentofBiomedicalEngi-neering,InstituteofUltrasoundEngineeringnMedicine,ChongqingMedicalUniversity,Chongqing40001
4、6,China) Abstract:Objective ToinvestigatetheeffectsofmouseamnioticfluidontheproliferationandapoptosisofmousehepatomacellineH22.Methods TreatedwiththemixtureofRPMI1640mediumand10%mouseamnioticfluidcollectedrespectivelyongestationalday11,13,15and17orRPMI1640mediumascontrol,theproliferationofH22cel
5、lswasdetectedwithMTTandtheapoptosisofH22cellswithAnnexinV-FITC;H22cellswereobservedmorphologicallyunderelectronmicroscope;Theexpressionsofproliferatingcellnuclearantigen(PCNA)andp53inH22cellswereinvestigatedwithimmunofluorescencestaining(S-Pmethod)andlasercon-focalmicroscopy.Results Amnioticfluid
6、significantlyinhibitedthecellproliferationaswellasthePCNAex-ressions,increasedthep53expressionsandinducedcellapoptosis.Theamnioticfluidcollectedongestationalday11wasthemosteffective,whilethatofgestationalday17wastheleasteffective(P<0•05),andtheireffectswerebothinatime-dependentmanner.Ther
7、ewasanegativecorrelationbetweenPCNAandp53(r=-0.808,P<0•01).Conclusion Amnioticfluidcollectedongestationalday11,13,15and17caninhibittheproliferationofH22cellsandinducetheirapoptosis.Theearlierpregnancyandthelong
