趋化因子Fractalkine协同M2型巨噬细胞对胰腺癌细胞株生物学功能的影响  

Effects of chemokine Fractalkine and M2 macrophages on biological functions of pancreatic cancer cell line

作　　者：刘惠娟 谢娟[2] 黄李雅[2] LIU Huijuan;XIE Juan;HUANG Liya(College of Clinical Medical,Ningxia Medical University,Yinchuan 750004,Ningxia Province,China;Department of Gastroenterology,General Hosptial of Ningxia Medical University,Yinchuan 750004,Ningxia Province,China)

机构地区：[1]宁夏医科大学临床医学院,宁夏银川750004 [2]宁夏医科大学总医院消化内科,宁夏银川750004

出　　处：《肿瘤》2019年第12期985-992,1010,共9页Tumor

基　　金：国家自然科学基金资助项目（编号：81860434)~~

摘　　要：目的:探讨趋化因子Fractalkine(FKN)联合肿瘤相关M2型巨噬细胞对人胰腺癌PANC-1细胞增殖、侵袭和迁移等生物学功能的影响。方法:将含FKN基因序列的重组慢病毒HBLV-h-FKN-GFP-PURO感染人胰腺癌PANC-1细胞,同时用佛波酯及白细胞介素4序贯诱导人白血病单核细胞THP-1成为M2型巨噬细胞,然后通过Transwell小室系统建立胰腺癌细胞与M2型巨噬细胞非接触式共培养模型。共培养后收集胰腺癌细胞,采用CCK-8法检测PANC-1细胞的增殖情况,Transwell小室法和划痕愈合实验分别检测细胞的侵袭和迁移能力。结果:HBLV-h-FKN-GFP-PURO感染人胰腺癌细胞PANC-1后,FKN蛋白表达水平较未感染对照组和空病毒感染组明显升高(P值均<0.001)。THP-1细胞经序贯诱导后成为高表达精氨酸酶Ⅰ(arginase-1,Arg-1)、低表达诱生型一氧化氮合酶(inducible nitric oxide synthase,iNOS)蛋白的M2型巨噬细胞(PArg-1<0.001,PiNOS<0.01)。过表达FKN后,PANC-1细胞趋化共培养M2型巨噬细胞的能力明显增强(P <0.001)。FKN过表达后,PANC-1细胞的增殖、侵袭和迁移能力均增强(P值均<0.001);而与M2型巨噬细胞共培养后,PANC-1细胞的增殖、侵袭和迁移能力均更加显著地增强(P值均<0.01);依据析因设计资料的双向分组方差分析结果提示,FKN与M2型巨噬细胞之间存在交互作用(P增殖<0.01,P侵袭<0.01,P迁移<0.05)。结论:趋化因子FKN能促使人胰腺癌PANC-1细胞趋化募集M2型巨噬细胞。FKN和M2型巨噬细胞均可增强PANC-1细胞的增殖、侵袭和迁移能力,且二者之间具有交互协同作用。Objective: To investigate the effects of cytokine Fractalkine(FKN) combined with M2-type macrophages on the proliferation, invasion, and migration of human pancreatic cancer PANC-1 cells.Methods: The recombinant lentivirus HBLV-h-FKN-GFP-PURO carrying FKN gene was constructed and infected into PANC-1 cells. THP-1 cells, a kind of monocytes of human leukemia, were induced into M2-type macrophages by phorbol 12-myristate 13-acetate(PMA) and interleukin-4(IL-4) in suquence. Then the non-contacting co-culture model of pancreatic cancer cells and M2-type macrophages with different expression level of FKN was established by Transwell chamber system. The proliferation, invasion and migration of human pancreatic cancer PANC-1 cells were detected by CCK-8 method, Transwell chamber test and wound-healing assay, respectievely.Results: As compared with the uninfected control and empty lentivirus infected groups, the expression level of FKN protein was significantly increased in human pancreatic cancer PANC-1 cells infected with recombinant lentivirus HBLV-h-FKN-GFP-PURO(both P < 0.001). THP-1 cells were successively induced to become M2-type macrophages with high expression of arginase-1(Arg-1) and low expression of inducible nitric oxide synthase(iNOS) protein(PArg-1 < 0.001, PiNOS < 0.01). The recruiting ability of PANC-1 cells to M2 type macrophages was enhanced after FKN over-expression(P < 0.001). The proliferation, invasion and migration abilities of PANC-1 cells were increased after FKN over-expression(all P < 0.001), and were more significantly increased after co-culture with M2-type macrophages(all P < 0.01). There were interactions between M2-type macrophages and FKN for proliferation, invasion and migration abilities of PANC-1 cells(P < 0.01, P < 0.01, P < 0.05).Conclusion: The chemokine FKN promotes the recruitment of PANC-1 cells to M2-type macrophages. Both FKN and M2-type macrophages can enhance the proliferation, invasion and migration abilities of PANC-1 cells, and there is synergistical interaction between

关 键 词：胰腺肿瘤 趋化因子FRACTALKINE M2型巨噬细胞 共同培养技术 

分 类 号：R735.9[医药卫生—肿瘤]