中华放射肿瘤学杂志
中華放射腫瘤學雜誌
중화방사종류학잡지
CHINESE JOURNAL OF RADIATION ONCOLOGY
2013年
2期
163-166
,共4页
高敏%张俊红%周云峰%周福祥%方绳权
高敏%張俊紅%週雲峰%週福祥%方繩權
고민%장준홍%주운봉%주복상%방승권
细胞系,肺肿瘤,人%X线照射%侵袭和转移%机制
細胞繫,肺腫瘤,人%X線照射%侵襲和轉移%機製
세포계,폐종류,인%X선조사%침습화전이%궤제
Cell line,lung neoplasms,human%X-ray irradiation%Invasion and metastasis%Mechanisms
目的 探讨x线照射后存活的人肺腺癌(A549)细胞侵袭和转移能力变化及机制.方法 用克隆形成实验检测X线照射前后放射敏感性变化,用MTT黏附实验、Transwell侵袭和迁移实验检测侵袭、转移能力.用SYBR Green荧光定量RT-PCR及蛋白印记法检测E-Cadherin、Vimentin、TGF-β1、MMP-2、MMP-9的mRNA及蛋白表达变化来探讨机制.结果 克隆形成实验结果显示X线照射后存活细胞对放射更加耐受(D0值比为0.94),其黏附、侵袭、迁移能力较照射前增加1.46、1.40、1.45倍.照射后细胞间隙增大,有多而长的伪足形成.照射后Vimentin、TGF-β1、MMP-2、MMP-9的mRNA表达均上调(为对照组的1.37、2.37、1.80、1.50倍),其蛋白表达也相应上调(为对照组的1.60、1.80、1.10、1.20倍),而E-Cadherin的mRNA及蛋白表达则下调(为对照组的59.4％、74.6％).结论 X线照射后A549细胞侵袭和转移能力较照射前明显增强,其机制可能与放射诱导TGF-β1表达进而促进上皮细胞间质化改变有关,还可能与放射诱导MMP-2、MMP-9表达增强有关.
目的 探討x線照射後存活的人肺腺癌(A549)細胞侵襲和轉移能力變化及機製.方法 用剋隆形成實驗檢測X線照射前後放射敏感性變化,用MTT黏附實驗、Transwell侵襲和遷移實驗檢測侵襲、轉移能力.用SYBR Green熒光定量RT-PCR及蛋白印記法檢測E-Cadherin、Vimentin、TGF-β1、MMP-2、MMP-9的mRNA及蛋白錶達變化來探討機製.結果 剋隆形成實驗結果顯示X線照射後存活細胞對放射更加耐受(D0值比為0.94),其黏附、侵襲、遷移能力較照射前增加1.46、1.40、1.45倍.照射後細胞間隙增大,有多而長的偽足形成.照射後Vimentin、TGF-β1、MMP-2、MMP-9的mRNA錶達均上調(為對照組的1.37、2.37、1.80、1.50倍),其蛋白錶達也相應上調(為對照組的1.60、1.80、1.10、1.20倍),而E-Cadherin的mRNA及蛋白錶達則下調(為對照組的59.4％、74.6％).結論 X線照射後A549細胞侵襲和轉移能力較照射前明顯增彊,其機製可能與放射誘導TGF-β1錶達進而促進上皮細胞間質化改變有關,還可能與放射誘導MMP-2、MMP-9錶達增彊有關.
목적 탐토x선조사후존활적인폐선암(A549)세포침습화전이능력변화급궤제.방법 용극륭형성실험검측X선조사전후방사민감성변화,용MTT점부실험、Transwell침습화천이실험검측침습、전이능력.용SYBR Green형광정량RT-PCR급단백인기법검측E-Cadherin、Vimentin、TGF-β1、MMP-2、MMP-9적mRNA급단백표체변화래탐토궤제.결과 극륭형성실험결과현시X선조사후존활세포대방사경가내수(D0치비위0.94),기점부、침습、천이능력교조사전증가1.46、1.40、1.45배.조사후세포간극증대,유다이장적위족형성.조사후Vimentin、TGF-β1、MMP-2、MMP-9적mRNA표체균상조(위대조조적1.37、2.37、1.80、1.50배),기단백표체야상응상조(위대조조적1.60、1.80、1.10、1.20배),이E-Cadherin적mRNA급단백표체칙하조(위대조조적59.4％、74.6％).결론 X선조사후A549세포침습화전이능력교조사전명현증강,기궤제가능여방사유도TGF-β1표체진이촉진상피세포간질화개변유관,환가능여방사유도MMP-2、MMP-9표체증강유관.
Objective To investigate the changes in the abilities of invasion and metastasis in surviving human lung adenocarcinoma (A549) cells after X-ray irradiation and the related mechanism.Methods The change in radiosensitivity after X-ray irradiation was determined by colony formation assay.The abilities of invasion and metastasis were evaluated by MTF adhesion assay and Transwell invasion and migration assay in vitro.The mRNA and protein expression of E-cadherin,vimentin,tumor growth factor (TGF)-β1,matrix metalloproteinase (MMP)-2,and MMP-9 was measured by real-time RT-PCR using SYBR Green fluorescence and Western blot.Results The colony formation assay showed that the surviving A549 cells after X-ray irradiation were more resistant to irradiation (ratio of D0 values =0.94).Their abilities of adhesion,invasion,and migration were significantly increased by 1.46 times,1.40 times,and 1.45 times,respectively.In addition,the surviving cells showed enlarged intercellular spaces and had many long pseudopodi.Compared with those in the control group,the mRNA expression levels of vimentin,TGF-β1,MMP-2,and MMP-9 of surviving cells were increased by 1.37 times,2.37 times,1.80 times,and 1.50 times,respectively,the protein expression levels of the above substances were increased by 1.60 times,1.80 times,1.10 times,and 1.20 times,respectively,and the mRNA and protein expression levels of E-cadherin were decreased by 59.4％ and 74.6％.Conclusions The surviving A549 cells after X-ray irradiation have significantly increased abilities of invasion and metastasis.This may be due to radiationinduced TGF-β1 expression increase that in turn promotes epithelial-mesenchymal transition and also due to radiation-induced MMP-2 and MMP-9 expression increase.
