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中华普通外科学文献(电子版) ›› 2015, Vol. 09 ›› Issue (03) : 198 -205. doi: 10.3877/cma.j.issn.1674-0793.2015.03.006

所属专题: 文献

论著

肿瘤相关巨噬细胞通过PI3K/Akt途径促进胰腺癌浸润迁移的研究
叶会霖1, 叶良涛1, 周雨1, 周泉波1, 林青1, 李志花1, 刘宜敏1, 陈汝福1,()   
  1. 1. 510120 广州,中山大学孙逸仙纪念医院胆胰外科
  • 收稿日期:2015-01-08 出版日期:2015-06-01
  • 通信作者: 陈汝福
  • 基金资助:
    国家自然科学基金资助项目(81000917;81370059)

Tumor-associated macrophages promoting invasion and migration of pancreatic cancer via PI3K/Akt signaling pathway

Huilin Ye1, Liangtao Ye1, Yu Zhou1, Quanbo Zhou1, Qing Lin1, Zhihua Li1, Yimin Liu1, Rufu Chen1,()   

  1. 1. Department of Pancreaticobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China
  • Received:2015-01-08 Published:2015-06-01
  • Corresponding author: Rufu Chen
  • About author:
    Corresponding author: Chen Rufu, Email:
引用本文:

叶会霖, 叶良涛, 周雨, 周泉波, 林青, 李志花, 刘宜敏, 陈汝福. 肿瘤相关巨噬细胞通过PI3K/Akt途径促进胰腺癌浸润迁移的研究[J/OL]. 中华普通外科学文献(电子版), 2015, 09(03): 198-205.

Huilin Ye, Liangtao Ye, Yu Zhou, Quanbo Zhou, Qing Lin, Zhihua Li, Yimin Liu, Rufu Chen. Tumor-associated macrophages promoting invasion and migration of pancreatic cancer via PI3K/Akt signaling pathway[J/OL]. Chinese Archives of General Surgery(Electronic Edition), 2015, 09(03): 198-205.

目的

研究PI3K/Akt信号通路对胰腺癌细胞PANC-1浸润迁移能力的影响,进一步探讨肿瘤相关巨噬细胞促进胰腺癌发生发展的分子机制。

方法

通过密度梯度离心法从健康成人外周血中分离单个核细胞,用IL-4体外诱导选择性激活的巨噬细胞(M2)。采用实时荧光定量PCR和Western blotting法检测胰腺癌PANC-1细胞PI3K、Akt mRNA和蛋白表达水平的变化,利用Transwell侵袭实验与划痕实验观察细胞浸润迁移能力的变化。

结果

体外模拟胰腺癌微环境,将胰腺癌PANC-1细胞与不同激活状态的巨噬细胞共培养,证明M2可显著上调胰腺癌PANC-1细胞PI3K、Akt的mRNA和蛋白水平,共培养20 h后可明显促进胰腺癌PANC-1细胞的浸润与迁移能力。

结论

肿瘤相关巨噬细胞可通过PI3K/Akt信号通路促进胰腺癌细胞的浸润与迁移。

Objective

To explore the influence of PI3K/Akt signaling pathway on invasive and migratory ability of pancreatic cancer cell line PANC-1, and further investigate the molecule mechanism of tumor-associated macrophages’promotion in the occurrence and development of pancreatic cancer.

Methods

Mononuclear cells were isolated from peripheral blood of healthy adults by density gradient centrifugation, and treated with IL-4 to obtain M2 in vitro. Expression of PI3K and Akt was evaluated by quantitative Real-time polymerase chain reaction and Western blotting. The ability of cellular invasion and migration was assessed by transwell chamber assay and wound healing assay.

Results

To simulate pancreatic cancer microenvironment, we co-cultured pancreatic cancer PANC-1 cells and Ua or M2. The mRNA and protein levels of PI3K and Akt were remarkably increased analyzing by qRT-PCR and western blotting. In addition, transwell chamber assays and wound healing assays revealed that alternatively activated macrophages significantly promoted the invasion and migration of PANC-1 cells in 20 h.

Conclusion

Tumor-associated macrophages may promote invasion and migration of pancreatic cancer cells via PI3K/Akt signaling pathway.

表1 PCR特异性引物序列
图1 不同激活状态的巨噬细胞光学显微镜下形态(400×)
图2 人外周血单个核细胞分离流式鉴定结果
图3 M2促进胰腺癌PANC-1细胞PI3K、Akt的mRNA转录和蛋白表达
图4 选择性巨噬细胞促进胰腺癌PANC-1细胞的横向迁移
图5 选择性巨噬细胞促进胰腺癌PANC-1细胞的纵向浸润
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