摘要
目的:施万细胞(Schwann cells,SCs)是周围神经系统中最主要的神经胶质细胞,在恶性肿瘤的发生、发展中起到了重要的促进作用。本研究旨在探讨SCs对胃癌细胞株HGC-27、BGC-823和AGS增殖和迁移的影响。方法:利用苏木精-伊红(hematoxylin-eosin,HE)染色和免疫组织化学(immunohistochemistry,IHC)染色观察胃癌组织中周围神经纤维与胃癌细胞之间的关系。采用无血清DMEM高糖培养基作为条件培养基(conditioned medium,CM),实验分为共培养组(CM组)和阴性对照(negative control,NC)组,CM组使用SCs CM培养胃癌细胞;NC组使用DMEM培养基培养胃癌细胞,并分别使用细胞计数试验盒8(cell counting kit-8,CCK-8)细胞增殖实验及细胞划痕试验检测SCs CM对胃癌细胞增殖及迁移能力的影响。利用Transwell小室构建胃癌细胞-SCs共培养体系,并将实验分为共培养组(Co-culture组)和NC组,Co-culture组Transwell小室上下层分别培养胃癌细胞及SCs;NC组Transwell小室上层培养胃癌细胞,下层为DMEM培养基,通过观察穿过小孔的胃癌细胞数量来检测SCs对胃癌细胞迁移能力的影响。利用生物信息学分析去分化前后SCs差异基因中的关键调控分子及其特异性受体,并检测两者在SCs及胃癌组织中的表达及定位。结果:HE及IHC结果显示胃癌细胞包绕癌巢周围神经纤维,甚至浸润至神经纤维内部,发生神经周围侵袭现象。CCK-8细胞增殖实验及细胞划痕试验显示:与NC组相比,CM组胃癌细胞的增殖及迁移能力均显著提高(均P<0.05)。Transwell共培养实验显示:与NC组相比,Co-culture组胃癌细胞的迁移能力显著提高(P<0.05)。生物信息学分析发现胶质细胞源性神经营养因子(glial cell derived neurotrophic factor,GDNF)在去分化SCs中表达显著增高。蛋白质印迹法显示CM培养后的SCs中GDNF表达增高,免疫荧光染色显示GDNF在胃癌组织中主要表达于周围神经纤维。GDNF家族受体α1(GDNF family receptor alpha 1,GFRα-1)蛋白在胃癌组织中呈高表达状态。结论:SCs可显著促进胃癌细胞增殖和迁移,GDNF-GFRα-1轴可能在其中发挥了重要的调控作用。
Objective:Schwann cells(SCs),the predominant glial cells in the peripheral nervous system,play a significant role in promoting the development and progression of malignant tumors.This study aims to investigate the effects of SCs on the proliferation and migration of gastric cancer cell lines HGC-27,BGC-823,and AGS.Methods:Hematoxylin-eosin(HE)staining and immunohistochemistry(IHC)were used to observe the relationship between peripheral nerve fibers and gastric cancer cells in tumor tissues.Primary rat SCs were cultured in serum-free high-glucose DMEM to produce conditioned medium(CM).The experiment was divided into a co-culture group(CM group)and a negative control group(NC group).The CM group used SCs CM to culture gastric cancer cells;in the NC group,the gastric cancer cells were cultured using DMEM medium.The effects of SCs CM on the proliferation and migration abilities of gastric cancer cells were respectively detected by the cell proliferation assay and the cell scratch assay of cell counting kit-8(CCK-8).The co-culture system of gastric cancer cells and SCs was constructed using Transwell chambers,the experiment was divided into a co-culture group and a NC group.In the co-culture group,the gastric cancer cells and SCs were cultured respectively in the upper and lower layers of the Transwell chamber;in the NC group,the gastric cancer cells were cultured in the upper layer of the Transwell chamber,and the lower layer was DMEM medium.The effect of SCs on the migration ability of gastric cancer cells was detected by counting the number of gastric cancer cells passing through the small hole.Bioinformatics analysis was performed to identify the key regulatory molecules and specific receptors associated with SC dedifferentiation,and their expression and localization in SCs and gastric cancer cells were examined.Results:HE and IHC results revealed that gastric cancer cells surrounded and even invaded the peripheral nerve fibers,indicating perineural invasion.The CCK-8 cell proliferation assay and cell scratch assay showed that:compared with the NC group,the proliferation and migration abilities of gastric cancer cells in the CM group were significantly enhanced(both P<0.05).The Transwell co-culture experiment showed that:compared with the NC group,the migration ability of gastric cancer cells in the co-culture group was significantly improved(P<0.05).Bioinformatics analysis indicated that glial cell-derived neurotrophic factor(GDNF)was significantly upregulated in dedifferentiated SCs.Western blotting confirmed increased GDNF expression in SCs cultured in serum-free high-glucose DMEM.Immunofluorescence staining showed that GDNF was predominantly expressed in peripheral nerve fibers within gastric cancer tissues.GDNF family receptor alpha 1(GFRα-1)was also found to be highly expressed in gastric cancer tissues.Conclusion:SCs significantly promote the proliferation and migration of gastric cancer cells,and the GDNF-GFRα-1 axis may play a crucial regulatory role in this process.
作者
兰东旭
谢鸣杰
仲铭洋
于卓群
吴寒
顾琪琪
LAN Dongxu;XIE Mingjie;ZHONG Mingyang;YU Zhuoqun;WU Han;GU Qiqi(Department of Gastrointestinal Surgery,Affiliated Hospital of Nantong University,Nantong 226001;Medical College of Nantong University,Nantong 226001,China)
出处
《临床与病理杂志》
2025年第2期127-137,共11页
Journal of Clinical and Pathological Research
基金
中国遗传资源保护与疾病防控教育部重点实验室开放课题(LPHGRD2024-003)。
关键词
施万细胞
胃癌
增殖
迁移
肿瘤微环境
Schwann cells
gastric cancer
proliferation
migration
tumor microenvironment
