Purpose To investigate the effect of purified culture of the colon cancer associated fibroblast ( CAF ), normal fibroblast ( NF ) on the proliferation and invasion of colon cancer cell line, and to study the the role of cancer cell' s micro-environment in the development of colorectal cancer. Methods CAF from colon carcinoma samples and normal colon fibroblast were collected to primary and pure cultivate. The morphology, proliferating and invasive of HCT-116 cell lines treated with CAF and fibroblast were observed. Results Purified cells of colon CAF were more dysplasia, loss of the contact and density inhibition was seen. CAF and NF cells were negative with cytokeratin CK( AE1/AE3 ) and positive with fibronectin, CAF was positive fora-SMA, but NF was negative. MTT results showed the proliferative activity of HCT-116 by adding the conditioned medium of CAF was higher than NF conditioned medium or the serum-free medium ( P <0. 05 ). Flow cytometry analysis showed that G2 and S phase cell percentage in HCT-116 co-cultured with CAF obviously increased . Under the action of CAF, the invasion ability of HCT-116 cells significantly enhanced ( P <0. 05 ). Conclusions Colon CAF may promote the proliferation and invasion of colon cancer cell lines HCT-116. CAF conditioned medium may contain some soluble substances which can promote the proliferation and invasion of HCT-116 cells.%目的 观察结肠癌相关成纤维细胞(cancer associated fibroblast,CAF)对结肠癌HCT-116细胞增殖及侵袭能力的影响,探讨癌细胞微环境在结直肠癌发生、发展过程中的作用.方法 采集结肠癌手术标本CAF及结肠正常成纤维细胞进行原代及纯化培养.观察HCT-116细胞被CAF及成纤维细胞作用后其形态学、增殖能力及侵袭能力的变化.结果 结肠CAF较成纤维细胞异型性明显,接触抑制和密度抑制丧失.结肠CAF及成纤维细胞免疫组化标记CK(AE1/AE3)均(-),成纤维细胞和CAFα-SMA均(+);CAF条件培养基组HCT-116增殖活性增高,CAF间接共培养HCT-116细胞G2期和S期所占比例明显增加,CAF作用下HCT-116细胞侵袭能力明显增强(P＜0.05).结论 CAF促进结肠癌HCT-116细胞增殖、侵袭及迁移;CAF条件培养基中可能存在可溶性活性物质,具有促进HCT-116细胞增殖侵袭能力的作用.
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