胰腺癌细胞与其基质成纤维细胞相互影响的研究

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。

The interaction between pancreatic cancer cells and fibroblasts

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.