SDF-1/CXCR4对大肠癌细胞株SW480增殖、迁移及侵袭的影响及意义

摘要

目的：探讨基质细胞衍生因子-1(SDF-1)及其特异性受体CXC趋化因子受体4（CXCR4）对大肠癌细胞SW480增殖、迁移及侵袭能力的影响及意义。方法取对数生长期大肠癌细胞SW480分为对照组(未经任何处理)、SDF-1组（加入100μg/L SDF-1）、SDF-1＋AMD3100混合组(向细胞中加入1 mg/L AMD3100，孵育2 h后加入100μg/L SDF-1)、AMD3100组（加入1 mg/L AMD3100）。免疫组化法检测SW480细胞中CXCR4蛋白表达情况；RT-PCR法检测SW480细胞中CXCR4 mRNA的表达情况，以及外源性SDF-1和AMD3100作用后CXCR4 mRNA表达水平的变化；MTT增殖实验、Transwell迁移及侵袭实验分别检测SDF-1以及AMD3100对SW480细胞增殖、迁移及侵袭能力的影响。结果 SW480细胞中CXCR4蛋白呈阳性表达（阳性率80%）。SW480细胞中有CXCR4 mRNA的表达，100μg/L SDF-1促使CXCR4 mRNA表达水平进一步上调，且能被1 mg/L AMD3100阻断。SDF-1组细胞增殖活性(0.847±0.039)高于对照组(0.624±0.011)和SDF-1＋AMD3100混合组(0.607±0.016)，AMD3100组(0.456±0.032)低于对照组和SDF-1＋AMD3100混合组(F=108.030，P＜0.05)。Transwell小室迁移及侵袭实验中SDF-1组穿膜细胞数(个：98.7±5.8、33.7±6.2)均多于对照组(21.0±2.2、6.1±2.3)、SDF-1＋AMD3100混合组(18.5±8.4、8.5±2.8)和AMD3100组(12.1±3.2、2.1±1.0)，后3组间比较差异无统计学意义。结论 SDF-1/CXCR4生物轴可促进大肠癌细胞SW480的增殖、迁移及侵袭。%Objective To discuss the influence and significance of stromal cell-derived factor 1 (SDF-1) and its specific receptor CXC chemokine receptor 4 (CXCR4) in proliferation, migration and invasion ability of SW480 colorectal cancer cells. Methods The colorectal cancer cell line SW480 in logarithmic phase was divided into four groups:control group (with no any processing), SDF-1 group (added 100μg/L SDF-1), SDF-1+1 mg/L AMD3100 mixed group (added 1 mg/L AMD3100 for 2 hours, then added 100μg/L SDF-1) and AMD3100 group (added 1 mg/L AMD3100). Immunohisto⁃chemistry method was used to detect the protein expression of CXCR4 in SW480 cells. The expression of CXCR4 mRNA in SW480 cells was detected by RT-PCR before and after SDF-1 and AMD3100 treatment. MTT assay and transwell chamber were used to test the changes of proliferation, migration and invasion ability of SW480 cells before and after SDF-1 and AMD3100 treatment. Results The result of immunohistochemistry showed that CXCR4 protein was expressed in SW480 cells (positive rate=80%). CXCR4 mRNA was expressed in SW480 cells. The expression of CXCR4 mRNA was up-regulat⁃ed by SDF-1(100μg/L), which could be inhibited by AMD3100 (1 mg/L). The proliferation activity was higher in SDF-1 group (0.847±0.039) compared to that in control group (0.624±0.011) and SDF-1+AMD3100 mixed group (0.607 ±0.016). The proliferation activity was lower in AMD3100 group (0.456 ± 0.031) than that in control group and SDF-1+AMD3100 mixed group (F=108.03, P<0.05). The number of transmembrane cells was more in SDF-1 group (98.7±5.8, 33.7±6.2) than that in control group (21.0±2.2, 6.1±2.3), SDF-1+1 mg/L AMD3100 mixed group (18.5±8.4, 8.5±2.8) and AMD3100 group (12.1±3.2, 2.1±1.0) detected by transwell chamber experiment. However, there were no statistical differences between three groups. Conclusion The biological axis SDF-1/CXCR4 can promote the proliferation, migration and invasion in colorectal cancer cell line SW480.