【作者】 张忠国； 牛瑞芳； 孙宝存； 郝希山；

【Author】 Zhang Zhong-Guo Niu Rui-Fang Sun Bao-Cun Hao Xi-Shan

【机构】 辽宁省肿瘤医院大肠科； 天津医科大学附属肿瘤医院；

【摘要】 目的:探讨结肠癌发生器官特异性转移的规律性, 研究CXCR1／CXCL8途经在转移性结肠癌瘤细胞归巢性肝转移中的作用。方法:应用流式细胞术分析不同转移潜能结肠癌瘤细胞株、肝转移性结肠癌原发灶及肝转移灶、高分化结肠腺癌原发灶、正常结肠牯膜和肝组织中CXCR1的表达及其差别;应用荧光定量PCR、MTT法、改良Boyden chamber法、激光扫描共聚焦显微镜,分析CXCR1／CXCL8 对肝转移性结肠癌瘤细胞CXCR1表达、瘤细胞生长、增殖性、侵袭能力、细胞骨架及定向运动能力的影响。结果:Lovo 细胞、colon205细胞膜表面CXCR1表达率分别为3．15％、 6．67％;lovo、colon205细胞经CXCL8刺激48kL,CXCR1 受体表达率50．45％和4．93％;lovo细胞经CXCL8(75ng／ ml)刺激前、后,CXCR1表达量(ct值)为20．83和 19．23,正常结肠粘膜、高分化结肠腺癌、低分化结肠腺癌中CXCR1的表达率分别为2．214％、6．652％、5．198％;正常肝组织、肝转移灶CXCR1的表达率为2．728％、22．62％; CXCL8刺激前、后Lovo细胞的增殖能力分别为3．27％、 24．17％;CXCL8刺激后,Lovo细胞F-actin表现出解聚- 聚合状态的改变,聚合程度呈时间性递增,高聚合状态表现在刺激后的72h;CXCR1／CXCL8作用前、后的lovo细胞侵袭数分别为74．2±13．28、6．39±1．8(P<0．01),趋化运动细胞数分别为89．67±6．16、18．53±5．59(P<0．05)。结论:CXCR1在自然状态下的表达与瘤细胞分化程度无关, 呈均一低表达状态;肝转移灶中,瘤细胞CXCR1转变为高表达状态; Lovo细胞中,CXCR1的表达受到其特异性配体的刺激,均一低表达状态也转变为高表达状态,并使其表现出转移灶瘤细胞的生长、增殖、运动特征;CXCL8／CXCR1 介导肝转移瘤细胞的器官特异性转移过程,使结肠癌肝转移表现出与淋巴细胞趋化性迁移相类似的归巢性转移特征。在归巢性转移过程中,配体-受体间具有器官特异性转移谱系特征,这些特征使得其对肿瘤细胞表现出“导航”效应,诱导转移瘤细胞的定向运动和转移灶形成。更多还原

【Abstract】 Objective: To explore the regularity of the organ-specific metastases on the metastatic colon cancer and to determine the chemotactic character of poor differentiated colon cancer with the liver metastasis. Methods: The expression of chemokin receptor CXCR1 in human colon cancer cell line with different metastatic potentials were examined using flow cytometric analysis, and the same comparison were also conducted in the primary colon cancer with distinct differentiation, the liver metastatic focus, the normal colon tissue and normal liver tissue. The effect of CXCR1/CXCL8 on the expression of CXCR1, cell growth and proliferation, the capability of invasive and directional migration, and re-regulation of cell cytoskeleton were analyzed respectively with real-time quantitative, MTT, modified Boyden chamber, and con-focal microscopy. Results: CXCR1 expression was low in lovo cell but elevated after the stimulation of different con- tent of the correspondingCXCL8. While these effects were limited to the lovo cell, the low level of CXCR1 in colon205, SW620, HCT-8 was not responding to the biological effects of IL-8. In the sample of tissues, including the normal colon mucus and liver tissue, the primary focus of high differentiated colon cancer and liver-metasatic colon cancer, the expression of receptor was low and no statistic significance with each other (p>0.05), while the result in the liver-metastatic focus was obviously rise, significantly different from other tissues (p<0.05). The invasive and migration cell numbers of lovo cell line was significantly different in the presence of CXCL8 or not and the former was higher than the later (p<0.05). It is observed that signaling through the interaction of CXCR1/CXCL8 mediates actin polymerization and pseudopodia formation, and subsequently induces chemotactic and invasive responses. Conclusion: The expression of CXCR1 is low and homogenenous in different kinds of colon cancer cells. While in the exist of CXCL8, the low expression of CXCR1 in lovo cell is changed into higher level situation and has the characteristics of liver metastatic colon cancer cells in cell growth, proliferation and directional migration. The expression of CXCR1 in liver metastatic focus is high and has the same model in vivo. It is concluded that the interaction of CXCR1/CXCL8 is related to the organ-specific metastases on the metastatic colon cancer. The process of homing liver metastases on colon cancer is share with the chemotactic migration of lymphocyte.更多还原