背景:胃癌(GC)是全球最常见的癌症之一。肿瘤微环境在肿瘤进展中起着重要作用。本研究旨在探讨癌相关成纤维细胞(CAFs)在GC中的作用及其机制。
方法:细胞活力,扩散,通过MTT评估入侵和迁移,EdU,transwell和伤口愈合试验,分别。球体形成测定用于评估细胞干细胞性。葡萄糖消耗,测量乳酸产生和ATP消耗以评估糖酵解。此外,通过qRT-PCR和Westernblot检测RNA和蛋白质的表达。无翼类型MMTV集成站点家族之间的交互,成员5A(WNT5A)和己糖激酶2(HK2)通过免疫共沉淀进行验证。建立异种移植模型以探讨CAFs对体内GC肿瘤生长的作用。
结果:CAFs促进了细胞增殖,转移,GC细胞的干性和糖酵解。WNT5A在CAF中上调,和CAFs增强了GC细胞中WNT5A的表达。GC细胞或CAF中WNT5A的敲低抑制了GC细胞的进展。此外,WNT5A促进HK2表达,HK2的过表达逆转了CAFs中WNT5A敲低对GC细胞的影响。此外,在CAFs中WNT5A的敲低抑制体内肿瘤生长。
结论:CAF来源的WNT5A通过调节HK2表达促进GC的进展。
BACKGROUND: Gastric cancer (GC) is one of the most common cancers worldwide. Tumor microenvironment plays an important role in tumor progression. This study aims to explore the role of cancer-associated fibroblasts (CAFs) in GC and the underlying mechanism.
METHODS: Cell viability, proliferation, invasion and migration were assessed by MTT, EdU, transwell and wound healing assays, respectively. Sphere formation assay was used to evaluate cell stemness. Glucose consumption, lactate production and ATP consumption were measured to assess glycolysis. In addition, The RNA and protein expression were detected by qRT-PCR and western blot. The interaction between wingless Type MMTV Integration Site Family, Member 5 A (WNT5A) and
hexokinase 2 (HK2) was verified by Co-immunoprecipitation. The xenograft model was established to explore the function of CAFs on GC tumor growth in vivo.
RESULTS: CAFs promoted the proliferation, metastasis, stemness and glycolysis of GC cells. WNT5A was upregulated in CAFs, and CAFs enhanced WNT5A expression in GC cells. Knockdown of WNT5A in either GC cells or CAFs repressed the progression of GC cells. In addition, WNT5A promoted HK2 expression, and overexpression of HK2 reversed the effect of WNT5A knockdown in CAFs on GC cells. Besides, knockdown of WNT5A in CAFs inhibits tumor growth in vivo.
CONCLUSIONS: CAF-derived WNT5A facilitates the progression of GC via regulating HK2 expression.