安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
ACTA UNIVERSITY MEDICINALIS ANHUI
2015年
4期
427-431
,共5页
苏娇娇%冯慧%姚玮艳%陈熹%张辰宇%王亚雷
囌嬌嬌%馮慧%姚瑋豔%陳熹%張辰宇%王亞雷
소교교%풍혜%요위염%진희%장신우%왕아뢰
胰腺癌%癌相关成纤维细胞%miRNAs%迁移
胰腺癌%癌相關成纖維細胞%miRNAs%遷移
이선암%암상관성섬유세포%miRNAs%천이
pancreatic cancer%cancer-associated fibroblasts%miRNAs%transwell
目的：观察胰腺癌细胞系BXPC-3、SW1990能否促进其基质中正常成纤维细胞( NFs)活化成癌相关成纤维细胞( CAFs)及其活化的可能机制,以及活化后的CAFs对BXPC-3、SW1990细胞迁移能力的影响。方法采用差异贴壁法分选出野生型小鼠C57胰腺组织中的NFs,后运用非接触式共培养方法将BXPC-3、SW1990与NFs共培养,共培养后采用免疫荧光方法检测共培养前后NFs中CAFs标志性蛋白α-平滑肌肌动蛋白(α-SMA)、成纤维细胞活化蛋白( FAP)的表达变化,以确定其是否活化,并且运用qRT-PCR法检测目前较公认的胰腺癌中升高的12种 miRNAs 在共培养前后的NFs中含量变化；采用 transwell 法观察活化前后的 NFs 对BXPC-3、SW1990迁移能力的影响。结果免疫荧光检测表明,与BXPC-3、SW1990共培养后, NFs中α-SMA、FAP表达均显著升高；同时,在所选取的12种miRNAs中,与BXPC-3、SW1990共培养后,NFs中miR-155含量均有明显升高。 BX-PC-3、SW1990在CAFs基质环境中迁移能力大大提高。结论胰腺癌细胞能够促进其周围NFs活化成CAFs,其活化可能与胰腺癌中某些特定的 miRNA分泌进入到 NFs中有关,活化后的CAFs又可以反过来促进胰腺癌细胞的迁移。
目的：觀察胰腺癌細胞繫BXPC-3、SW1990能否促進其基質中正常成纖維細胞( NFs)活化成癌相關成纖維細胞( CAFs)及其活化的可能機製,以及活化後的CAFs對BXPC-3、SW1990細胞遷移能力的影響。方法採用差異貼壁法分選齣野生型小鼠C57胰腺組織中的NFs,後運用非接觸式共培養方法將BXPC-3、SW1990與NFs共培養,共培養後採用免疫熒光方法檢測共培養前後NFs中CAFs標誌性蛋白α-平滑肌肌動蛋白(α-SMA)、成纖維細胞活化蛋白( FAP)的錶達變化,以確定其是否活化,併且運用qRT-PCR法檢測目前較公認的胰腺癌中升高的12種 miRNAs 在共培養前後的NFs中含量變化；採用 transwell 法觀察活化前後的 NFs 對BXPC-3、SW1990遷移能力的影響。結果免疫熒光檢測錶明,與BXPC-3、SW1990共培養後, NFs中α-SMA、FAP錶達均顯著升高；同時,在所選取的12種miRNAs中,與BXPC-3、SW1990共培養後,NFs中miR-155含量均有明顯升高。 BX-PC-3、SW1990在CAFs基質環境中遷移能力大大提高。結論胰腺癌細胞能夠促進其週圍NFs活化成CAFs,其活化可能與胰腺癌中某些特定的 miRNA分泌進入到 NFs中有關,活化後的CAFs又可以反過來促進胰腺癌細胞的遷移。
목적：관찰이선암세포계BXPC-3、SW1990능부촉진기기질중정상성섬유세포( NFs)활화성암상관성섬유세포( CAFs)급기활화적가능궤제,이급활화후적CAFs대BXPC-3、SW1990세포천이능력적영향。방법채용차이첩벽법분선출야생형소서C57이선조직중적NFs,후운용비접촉식공배양방법장BXPC-3、SW1990여NFs공배양,공배양후채용면역형광방법검측공배양전후NFs중CAFs표지성단백α-평활기기동단백(α-SMA)、성섬유세포활화단백( FAP)적표체변화,이학정기시부활화,병차운용qRT-PCR법검측목전교공인적이선암중승고적12충 miRNAs 재공배양전후적NFs중함량변화；채용 transwell 법관찰활화전후적 NFs 대BXPC-3、SW1990천이능력적영향。결과면역형광검측표명,여BXPC-3、SW1990공배양후, NFs중α-SMA、FAP표체균현저승고；동시,재소선취적12충miRNAs중,여BXPC-3、SW1990공배양후,NFs중miR-155함량균유명현승고。 BX-PC-3、SW1990재CAFs기질배경중천이능력대대제고。결론이선암세포능구촉진기주위NFs활화성CAFs,기활화가능여이선암중모사특정적 miRNA분비진입도 NFs중유관,활화후적CAFs우가이반과래촉진이선암세포적천이。
Objective To investigate whether the pancreatic cancer cells BXPC-3,SW1990 can activate normal fi-broblasts( NFs) into cancer-associated fibroblasts( CAFs) and the possible mechanism and whether CAFs can moti-vate the migration of the cancer cells. Methods NFs were isolated from mouse pancreas tissues of wild C57,then NFs were cultured with BXPC-3 or SW1990 cells by indirect co-cultures, and detected the content of the specific proteins α-SMA, FAP in fibroblasts by immunofluorescence assay, then detected the content of the 12 different kinds of miRNAs in fibroblasts which were regarded as overexpressed in pancreatic cancer by qRT-PCR, and esti-mated the effect of the different fibroblasts( NFs,CAFs) on cancer cell migration by transwell. Results After coc-ultered with BXPC-3 or SW1990 cells, the α-SMA and FAP level in fibroblasts increased dramatically. Among the selected 12 kinds of miRNAs, after cocultered with BXPC-3 or SW1990 cells,the content of miR-155 in fibroblasts raised significantly. The migration of BXPC-3 ,SW1990 cells increased dramatically in the environment of CAFs compared with the environment of NFs. Conclusion BXPC-3 or SW1990 cells can activate NFs into CAFs, and miRNAs may play an important role in the process,then CAFs can reversely motivate the migration of BXPC-3 or SW1990 cells.
