体外胃癌间质成纤维细胞模型建立和生物学特性初步研究

Establishment of vitro cell model of human gastric mucosal fibroblasts and preliminary exploration of its biological features

目的采用原代培养胃间质成纤维细胞(human gastric mucosal fibroblasts,HGMF),构建体外胃癌间质成纤维细胞模型,研究其生物学特性。方法将调整培养条件的胃癌细胞系MKN-45的培养上清液加入HGMF中培养,使其发生转化。MTT法检测其细胞增殖,ELISA方法测定其肝细胞生长因子(hepatocyte growth factor,HGF)、转化生长因子-β1(transforming growth factor beta 1,TGF-β1)、趋化因子9(chemokine ligand 9,CXCL9)、趋化因子5(chemokine ligand 5,CCL5)的表达变化,RT-PCR检测其端粒酶活性,BSA法检测蛋白合成能力。结果胃癌细胞培养上清液与HGMF共培养可使其发生转化,转化HGMF细胞增殖能力(0.860±0.055)较HGMF(0.639±0.074)增强,差异有统计学意义(P<0.05),HGF、TGF-β1、CCL5和CXCL9表达增强,差异有统计学意义(P<0.01),端粒酶活性增高,差异有统计学意义(P<0.01),蛋白质合成能力增强,差异有统计学意义(P<0.01)。结论转化成纤维细胞与亲代细胞相比具有更强的生物学活性和分裂活性,蛋白质合成增强,能表达和分泌更多的与胃癌细胞生长相关的生长因子和趋化因子。

Objective To establish vitro cell model of human gastric mucosal fibroblasts （ HGMF） in gastric cancer and ex-plore its biological features. Methods HGMF were cultured with the regulated supernatant of MKN-15 cells, facilitating its transforma-tion. Cell proliferation was detected by MTT assay. The expressions of hepatocyte growth factor（ HGF） , transforming growth factor beta 1 （TGF-β1 ） , chemokine ligand 5 （ CCL5） and chemokine ligand 9（ CXCL9） were detected by ELISA. The activity of telomerase and the ability to synthesize proteins were detected by RT-PCR and BSA assay respectively. Results Supernatant of MKN-15 could induce HGMF transformation. The ability of cell proliferation was enhanced （P 〈 0 . 05） , and the expression levels of HGF, TG F-β1, CCL5 and CXCL9 were also increased （ P 〈 0. 01 ）. Telomerase activity and the ability to synthesize proteins were also enhanced respectively （P 〈0.01）. Conclusion Detection results suggested that the transformed HGMF seemed to display higher biological activity, as com-pared with that of the parent cells, with stronger proliferation activity and higher protein synthesis, and the HGMF could express and se-crete more growth factor and chemokines associated with the growth of gastric cancer cells.