胃癌相关成纤维细胞及人胃黏膜成纤维细胞生物学特性比较

Comparison of biological features of gastric cancer-associated fibroblasts and human gastric mucosa fibroblasts

目的 观察胃癌相关成纤维细胞（GCAF）及人胃黏膜成纤维细胞（HGMF）生物学特性的差异.方法 植块法原代培养和鉴定GCAF及HGMF.选择第3代的GCAF及HGMF,观察两种细胞光镜和透射电镜下的形态特点 CCK-8法检测细胞增殖活性 BCA法检测细胞总蛋白 流式细胞术检测细胞周期.每组实验重复3次,计算平均值进行比较.结果 与HGMF对比,光镜下GCAF胞体变大,细胞不规则,多见大胞体细胞、双核及多核细胞 电镜下可见畸形核,多个核仁,核浆比增加,线粒体、滑面内质网增多.GCAF增殖活性在第2、4、5、6、7天明显增强（P＜0.05） 1×106个GCAF和HGMF蛋白浓度分别为（1.0018±0.0940）g/L和（0.7688±0.0441）g/L（P＜0.01） GCAF和HGMF的增殖指数分别为（42.0200±2.7314）%和（35.2520±4.4368）%（P＜0.05） 两种细胞的SPF分别为（19.5040±2.7944）%和（9.3180±2.2068）%（P＜0.01）.结论 HGMF和GCAF不仅在形态学上存在较大差异,GCAF的增殖能力、蛋白合成能力以及细胞周期活性均高于HGMF.

Objective To investigate the biologic characteristic differences between gastric cancerassociated fibroblasts （GCAF） and human gastric mucosa fibroblasts （HGMF）. Methods The fibroblasts were cultured primarily by outgrowth method and identified by immunohistochemistry. Third-generation cells were chosen to be analyzed. Their differences in biological features were compared by light microscopy, electron microscopy, and CCK-8 assay. The ability to synthesize proteins and cell cycle were detected by BCA Protein Assay Kit and flow cytometry respectively. Each experiment was repeated three times and the average was calculated. Results Compared with HGMF, the both of GCAF was larger and irregular with conjugate nuclei or multinucleate cells under the light microscopy. Deformed nuclei and multiple nuclei could be found in GCAF under the electron microscopy with elevated karyoplasmic ratio and increased mitochondria smooth endoplasmic reticulum. The ability of proliferation and protein synthesis in GCAF was significantly enhanced as compared with HGMF （P 〈0. 01 ）, and both proliferation index （P 〈0. 05 ） and percentage of S phase cells （ P 〈 0. 01 ） in GCAF were significantly higher than HGMF. Conclusion There were not only obviously differences in morphology, but also ability of proliferation, protein synthesis and cell cycle activity in GCAF were significantly higher than HGMF.