【摘要】 目的 探讨多配体蛋白聚糖1 (SDC-1)在胃癌患者癌组织和正常胃黏膜组织中的表达及其在胃癌细胞系中的生物学功能。方法 采用免疫组织化学染色（IHC）与GEPIA数据库分析SDC-1在胃癌组织与正常胃组织中的差异化表达水平，TIMER数据库分析胃癌肿瘤微环境中免疫细胞浸润与SDC-1表达的关系，蛋白印迹法验证SDC-1在胃癌细胞系中的表达水平；通过质粒转染构建SDC-1敲除/过表达的胃癌细胞系，应用细胞增殖和细胞毒性、5-乙炔基-2’脱氧尿嘧啶核苷染色（EdU）、集落形成、细胞划痕和Transwell实验观察细胞增殖、迁移和侵袭能力的变化；EdU和细胞划痕实验观察共培养后细胞增殖和迁移能力的变化。结果 IHC结果显示，胃癌患者癌组织中SDC-1表达水平高于癌旁对照正常组织（P<0.05）;GEPIA数据库中分析结果显示，相比正常胃黏膜组织，SDC-1在胃癌组织中呈高表达（P<0.05）。TIMER数据库分析提示，SDC-1的表达水平与CD8+T细胞、CD4+幼稚T细胞、效应B细胞、自然杀伤细胞、M1巨噬细胞呈负相关关系，与肿瘤相关成纤维细胞呈正相关关系（P<0.05）；蛋白印迹法结果显示，相比GES-1细胞，SDC-1在胃癌细胞中呈高表达；CCK8、EdU、集落形成、细胞划痕和Transwell实验表明，与对照组比较，干扰SDC-1后胃癌HGC-27细胞的增殖、迁移、集落形成和侵袭能力降低（P<0.05）,MKN-45胃癌细胞的增殖和侵袭能力显著降低（P<0.05）；而过表达SDC-1后胃癌AGS细胞的增殖、迁移、集落形成和侵袭能力提高（P<0.05）；与过表达SDC-1的AGS细胞共培养后，AGS细胞的增殖和迁移能力增加（P<0.05）。结论 SDC-1在胃癌患者癌组织中呈高表达，高表达的SDC-1与胃癌肿瘤微环境免疫细胞浸润相关；干扰SDC-1的表达可抑制胃癌细胞功能的发挥，过表达及共培养体系脱落型SDC-1能够促进胃癌细胞生长和迁移。更多还原

【Abstract】 Objective To investigate the expression of syndecan-1(SDC-1) in cancerous and normal gastric mucosal tissues from gastric cancer patients and its biological function in gastric cancer cell lines.Methods Differential expression levels of SDC-1 in gastric cancer tissues and normal tissues were analyzed by immunohistochemical staining(IHC) with GEPIA database; TIMER database was used to analyze the relationship between immune cell infiltration and SDC-1 in the microenvironment of gastric cancer;Western blotting was used to validate the expression level of SDC-1 in the gastric cancer cell lines. These cell lines with interference/overexpression of SDC-1 were constructed by plasmid transfection. CCK8, 5-ethynyl-2’-deoxyuridine(EdU), colony formation, scratch and Transwell assays were applied to observe the changes in cell proliferation, migration and invasion ability. EdU and scratch assays were used to observe the changes in cell proliferation and migration ability after co-culture. Results IHC results showed that the expression level of SDC-1 in cancer tissues of gastric cancer patients was higher than that in the normal tissues of paracancerous control(P<0.05). The analysis results in the GEPIA database showed that SDC-1 was highly expressed in gastric cancer tissues compared with that in normal gastric tissues(P<0.05). The analysis in the TIMER database suggested that the expression level of SDC-1 was negatively correlated with CD8+ T cells, CD4+ naive T cells, effector B cells, natural killer cells and M1 macrophages, and positively correlated with tumor-associated fibroblasts(P<0.05). Western blotting results showed that SDC-1 was highly expressed in gastric cancer cells compared with GES-1 cells; CCK8, EdU, colony formation, scratch and Transwell assays showed that the proliferation, migration, colony formation and invasion abilities of gastric cancer HGC-27 cells decreased after being interfered with SDC-1 compared with the control group(P<0.05), and the proliferation and invasion ability of MKN-45 were significantly reduced(P<0.05); whereas the proliferation, migration, colony formation and invasion ability of gastric cancer AGS cells increased after and overexpression of SDC-1(P<0.05), and, after co-culture with AGS overexpressed with SDC-1, the proliferation and migration ability of AGS cells increased(P<0.05). Conclusion SDC-1 is highly expressed in cancer tissues of gastric cancer patients, and the high expression of SDC-1 correlates with the infiltration of immune cells in the microenvironment of gastric cancer tumors; interfering SDC-1 can inhibit the function of gastric cancer cells, and an overexpression and co-culture system of shed SDC-1 can promote the growth and migration of gastric cancer cells.更多还原