趋化因子CXCL14在不同转移潜能结直肠癌细胞中的表达及其与血管生成的关系

Expressions of chemokine CXCL14 in colorectal cancer cells of different metastasis potentials and its association with angiogenesis

背景与目的:CXC趋化因子配体14(CXCL14)是一种与肿瘤细胞迁移密切相关的趋化因子,与多种恶性肿瘤的发生、进展有关。本研究旨在探讨CXCL14在结直肠癌细胞中的表达以及对血管生成的影响。方法:用RT-PCR与Western blot法分别检测CXCL14基因与蛋白在不同结直肠癌细胞株(HT-29、WiDr、CaCo-2、Colo-320)中的表达,以及转染CXCL14 siRNA后的影响。分别用WST-1法、Transwell小室实验和血管生成实验检测不同浓度CXCL14或同时添加CXCL14抗体对人脐静脉内皮细胞(HUVEC)增殖、迁移、血管生成能力的影响,并观察HUVEC与不同结直肠癌细胞株共培养后血管生成能力的差异。结果:CXCL14基因和蛋白表达于高肝转移潜能结直肠癌细胞株(HT-29、WiDr),而低肝转移潜能结直肠癌细胞株(CaCo-2、Colo-320)中无表达。转染CXCL14 siRNA后,HT-29和WiDr细胞CXCL14蛋白的表达被明显抑制,而CaCo-2、Colo-320细胞无变化。CXCL14作用后,HUVEC的增殖、迁移、血管生成能力明显增强,并呈浓度依赖性(均P<0.05),但以上作用均被同时添加CXCL14抗体取消(均P<0.05)。HUVEC与无CXCL14表达的CaCo-2细胞共培养后的血管生成数量明显低于与表达CXCL14的HT-29细胞共培养后的血管生成数量(P<0.05),但与转染CXCL14 siRNA的HT-29细胞共培养后的血管生成数量无明显差异(P>0.05)。结论:CXCL14在高肝转移潜能的结直肠癌细胞中表达,其可能通过增强血管内皮细胞的增殖与迁移能力来增加血管新生,从而促进了结直肠癌的肝转移。

Background and Aims:C-X-C motif chemokine ligand 14 (CXCL14) is a chemokine that induces migration of tumor cells,and closely related to the occurrence and development of various malignant tumors.This study was conducted to investigate the expression of CXCL14 in colorectal cancer cells and its influence on angiogenesis.Methods:The expressions of CXCL14 mRNA and protein in different colorectal cancer cell lines (HT-29,WiDr,CaCo-2 and Colo-320) were detected by RT-PCR and Western blot analysis respectively,and the effects of CXCL14 siRNA transfection on these cells were also observed.The effects of different concentrations of CXCL14 alone or with simultaneous addition of CXCL14 antibodies on proliferation,migration and angiogenesis abilities of human umbilical vein endothelial cells (HUVECs) were measured by WST-1 assay,Transwell migration assay and angiogenesis assay,respectively.The differences in angiogenesis ability of HUVECs after co-culture with different colorectal cancer cell lines were also observed.Results:The CXCL14 mRNA and protein were expressed in colorectal cancer cell lines with high hepatic metastasis potential (HT-29 and WiDr),but were absent in colorectal cancer cell lines with low hepatic metastasis potential (CaCo-2 and Colo-320).After CXCL14 siRNA transfection,the CXCL14 protein expressions in HT-29 and WiDr cells were remarkably decreased,and were remained unchanged in CaCo-2 and Colo-320 cells.After CXCL14 treatment,the proliferation,migration and angiogenesis abilities of HUVECs were all significantly enhanced with a concentration dependent manner (all P<0.05),but these effects were all abolished by simultaneous addition of CXCL14 antibodies (all P<0.05).The number of vessel formation of HUVECs after co-culture with CaCo-2 cells that didnot express CXCL14 was significantly lower than that of HUVECs after co-culture with HT-29 cells that express CXCL14 (P<0.05),but showed no significant difference with that of HUVECs after co-culture with HT-29 cells transfected with CXCL14 siRNA (P>0.05).Conclusion:CXCL14 is expressed in colorectal cancer cells with high liver metastasis potential.It may increase the angiogenesis through enhancing the proliferation and migration abilities of vascular endothelial cells,and thereby promote the metastasis of colorectal cancer to the liver.