基质细胞衍生因子/趋化因子受体4轴参与胰腺癌细胞侵袭的体外实验研究

目的 探讨胰腺癌细胞株趋化因子受体(CXCR)4的表达及其与增殖、侵袭和粘附的关系.方法 采用RT-PCR检测3个胰腺癌细胞株中CXCR4 mRNA的表达;Western印迹法检测胰腺癌细胞株中CXCR4的蛋白表达;共聚焦显微镜检测基质细胞衍生因子(SDF)-1α作用下AsPC-1细胞内钙荧光强度变化;MTT法检测细胞增殖;细胞体外粘附试验及Transwell体外侵袭实验观察胰腺癌细胞的粘附及侵袭能力.结果 3个胰腺癌细胞株均不同程度地存在CXCR4 mRNA及蛋白表达,AsPC-1细胞表达最强,而SW1990表达最弱;CXCR4功能性表达于AsPC-1细胞;SDF-1α不同程度的促进3种胰腺癌细胞的增殖、侵袭及粘附,尤以AsPC-1细胞最为明显,而SW1990细胞最弱;AMD3100可抑制SDF-1α所致的促增殖、侵袭及粘附作用.结论 胰腺癌细胞表达CXCR4mRNA及蛋白,SDF-1α通过与CXCR4作用影响胰腺癌细胞的侵袭转移,该效应与CXCR4表达程度密切相关.

In vitro study on stromal cell derived factorl/chemokine receptor 4 axis that involved in the invasion of human pancreatic cancer cells

Objective To investigate the expression of chemokine receptor (CXCR)4 in human pancreatic cancer cell lines,and its association with proliferation,adhesion and invasion of pancreatic cancer cells.Methods The CXCR4 mRNA and protein in three pancreatic cancer cell lines were detected by RT-PCR and Western blotting,respectively.Confocal microscopy was used to detect the fluorescence intensity induced by SDF-lα in AsPC-1 cells.MTT test was performed to study the proliferation of pancreatic cancer cells.The invasive ability of pancreatic cell lines was determined by transwell invasion assay kit and the adhesive ability was detected by cell adhesive test in vitro.Results There were expressions of CXCR4 mRNA and protein in different extent in three pancreatic cancer cell lines.The strong expression was seen in AsPC-1 cell line,but weak expression in SW1990 cell line.The CXCR4 was functional expressed on AsPC-1 ceils.SDF-1α improved the proliferation,adhesion and invasion of three pancreatic cancer cell lines,especially in AsPC-1 cell line,while the proliferation in SW1990 cell line was weak.But all above effects of the SDF-1α could be inhibited by AMD3100.Conclasions CXCR4 mRNA and protein were expressed in pancreatic cancer cell lines.The efficacy that SDF-1 can increase the invasive ability of pancreatic cancer ceils through SDF-1/CXCR4 axis is closely related to the expression of CXCR4.