衣霉素對(duì)順鉑誘導(dǎo)人宮頸癌細(xì)胞凋亡的影響.pdf

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1、MedJChinPLA，Vo1．38，No．4，April1，2013283論著衣霉素對(duì)順鉑誘導(dǎo)人官頸癌細(xì)胞凋亡的影響徐冶，李質(zhì)馨，曹慧玲，于洋，劉師兵，王曉軍[摘要]目的探討內(nèi)質(zhì)網(wǎng)應(yīng)激在順鉑誘導(dǎo)人宮頸癌HeLa~胞凋亡中的作用。方法取人宮頸癌HeLa細(xì)胞作為研究對(duì)象，實(shí)驗(yàn)分為衣霉素(Smg／L)組、順鉑(6mg／L)組、衣霉素(5mg／L)與Jt~fl(6mg／L)聯(lián)合給藥組、陰性對(duì)照組，處理12h后采用MTT法觀察細(xì)胞生長(zhǎng)情況，Hoechst熒光染色檢測(cè)細(xì)胞核形態(tài)的變化，免疫印跡法檢測(cè)凋亡蛋白Caspase一

2、3、Caspase．4活化情況，間接免疫熒光法檢測(cè)蛋白質(zhì)二硫鍵異構(gòu)酶(PDI)、磷酸化H2AX組蛋白(．H2AX)的表達(dá)變化。結(jié)果MTT法檢測(cè)顯示衣霉素組、順鉑組、聯(lián)合給藥組和陰性對(duì)照組的細(xì)胞增殖抑制率分別為2．65％±2．7l％、19．60％4-4．34％、44．69％±7．07％、0，除衣霉素組與對(duì)照組外其余各組間兩兩比較差異均有統(tǒng)計(jì)學(xué)意義(P

3、組核碎裂細(xì)胞比例(44．s％±5．1％)顯著高于順鉑組(22．7％±3．9％，P<0．05)。免疫印跡法檢測(cè)顯示，順鉑組裂解Caspase+3、Caspase一4蛋白表達(dá)明顯高于對(duì)照組和衣霉素組，但顯著低于聯(lián)合給藥組(P

4、出現(xiàn)明顯綠色熒光，但兩組無(wú)明顯差異。結(jié)論衣霉素可增加內(nèi)質(zhì)網(wǎng)應(yīng)激水平，促進(jìn)順鉑誘導(dǎo)HeLa細(xì)胞發(fā)生凋亡。[關(guān)鍵詞]宮頸腫瘤；衣霉素；順鉑；內(nèi)質(zhì)網(wǎng)應(yīng)激；細(xì)胞凋亡[中圖分類號(hào)]R737．33[文獻(xiàn)標(biāo)識(shí)碼]A[文章編號(hào)]0577．7402(2013)04．0283．05EffectoftunicamycinoncisplatininducedapoptosisofHeLacellsXUYe，LIZhi—xin。，CAOHui-ling，YUYang，LIUShi—bing，WANGXiao—junDepartmentof

5、HistologyandEmbryology,2MedicalResearchLaboratory,JilinMedicalCollege,]ilinCity,Jilin132013,China’Correspondingautho~E—mail：Izx一62@163．cornThiswork1．~assupportedbytheNaturalScienceFoundationofJilinProvincef2Ol015240)[Abstract]ObjectiveToinvestigatetheeffectof

6、endoplasmicreticulumstress(ERstress)incisplatin—inducedapoptosisofhumancervicalcancerHeLacells．MethodsHeLacellswereusedasthestudyobjectwhichweredividedintofourgroups：TUNI(Smg／L)group，cisplatin(6mg／L)group,TuNI(5mg／L)+cisplatin(6mg／L)group,andnegativecontrolgr

7、oup(nodrugtreatment)．MTTassaywasemployedtoexaminethegrowthstatusofthecells．Hoechststainingwasusedtoobservethemorphologicalchangeinthenucleus．ImmunoblottingwasusedtodetecttheactivationofapoptoticproteinsJcaspase一3andcaspase一4．Indirectimmunofluorescencewasusedt

8、oassesstheexpressionoftheproteindisulfdeisomerase(PDI)andphosphorylatedhistoneH2AX(^y—H2AX)．ResultsMTTassayshowedthatthegrowthinhibitionrateswere2．65％±2．71％J19．60％±4．34％J44．69％±7．07％and0％