[摘要] 目的：探讨肿瘤相关巨噬细胞（tumor-associated macrophage，TAM）对胃癌MGC-803 细胞增殖、迁移、侵袭、凋亡的影响及其可能的作用机制。方法：体外培养人单核细胞株THP-1，加入佛波酯（PMA）和IL-4 后，用ELISA法检测细胞培养上清液中IL-12、IL-10 的水平。取对数生长期MGC-803 细胞和M2 型TAM，根据培养方式不同分为单独细胞培养组、非接触共培养组和接触共培养组，用MTT法、Transwell小室法分别检测MGC-803 细胞的增殖、迁移和侵袭能力，用AnnexinV-FITC/PI 双染流式细胞术检测MGC-803 细胞的凋亡和细胞周期变化，用qPCR、Western blotting 分别检测MGC-803 细胞中MMP-9、MMP-2 mRNA和蛋白的表达水平。结果：与PMA组相比，PMA+IL-4 组细胞上清液中IL-12 水平显著降低、IL-10 水平显著升高（均P<0.05），成功将THP-1细胞诱导分化为M2型TAM。与单独细胞培养组相比，非接触共培养组、接触共培养组（: 1）MGC-803细胞的增殖率显著升高（均P<0.05）；（2）细胞的迁移、侵袭数量升高（均P<0.05）；（3）细胞凋亡率显著降低（均P<0.05）；（4）S、G2 期细胞的数量升高、G1 期数量降低（均P<0.05）；（5）细胞中MMP-9、MMP-2 mRNA和蛋白的表达水平均显著升高（均P<0.05）。结论：TAM可促进胃癌MGC-803 细胞增殖、迁移和侵袭，解除细胞G1期阻滞，减少细胞凋亡，其机制可能与上调胃癌细胞MMP-9、MMP-2表达有关。

[Abstract] Objective: To investigate the effects of tumor-associated macrophages (TAM) on proliferation, migration, invation and apoptosis of gastric cancer MGC-803 cells and the possible mechanisms. Methods: Human monocyte THP-1 was cultured in vitro. After being added with PMA and IL-4, the levels of interleukin-12 (IL-12) and interleukin-10 (IL-10) in cell supernatant were detected by enzyme-linked immunosorbent assay (ELISA). MGC-803 cells at logarithmic phase and M2-type TAM cells were divided into single cell culture group, non-contact co-culture group and contact co-culture group according to different culture methods. MTT assay was used to detect the proliferation of MGC-803 cells, Transwell assay was used to detect cell migration and invasion, and Annexin V-FITC/PI staining flow cytometry was used to examine the apoptosis and cell cycle changes of MGC-803 cells; In addition, the mRNA and protein expressions of matrix metalloproteinase-9 (MMP-9) and MMP-2 were detected by Real-time fluorescence quantitative PCR (qPCR)and Western blotting. Results: Compared with PMA group, the level of IL-12 in cell supernatant of PMA+IL-4 group decreased significantly while the level of IL-10 increased significantly (all P<0.05), indicating THP-1 cells were successfully induced to differentiate into M2-type TAM. Compared with the single cell culture group, the non-contact co-culture group and the contact co-culture group exhibited: (1) significantly increased proliferation rate of MGC-803 cells (P<0.05)；(2) increased number of migrated and invaded cells (all P < 0.05)；(3) significantly decreased apoptotic rate (P<0.05)；(4) increased proportion of S, G2 phase cells and decreased proportion of G1 phase cells (all P<0.05)；and (5) significantly increased mRNA and protein expressions of MMP-9 and MMP-2 (all P<0.05).Conclusion: TAM can promote the proliferation, migration and invasion of gastric cancer MGC-803 cells, relieve G1 phase arrest and reduce cell apoptosis, which may be related to the up-regulation of MMP-9 and MMP-2 expression in gastric cancer cells.

辽宁省科学技术计划资助项目（No. 2015020377）