Abstract：
BACKGROUND: Cancer-associated fibroblasts (CAFs) are major components of the tumor microenvironment (TME). Hypoxic TME is known to promote tumor progression. However, how a hypoxic condition regulates CAFs remains elusive.
METHODS: To investigate the underlying mechanism involved in the regulation of gastric cancer (GC) progression by hypoxic CAFs, we performed secretome profiling. Normoxic or hypoxic CAFs conditioned media (CM) were filter-concentrated and in-gel trypsin digested. Resulting peptides were analyzed with LC-MS/MS.
RESULTS: We observed that CM derived from hypoxic CAFs could promote migration of a panel of GC cell lines (AGS, SNU668, SNU638). Mass spectrometry analysis of hypoxic or normoxic CAFs CM identified 1595 proteins, of which 19 proteins (10 upregulated and 9 downregulated) were differentially expressed in the hypoxic secretome. We focused on COL4A2, whose expression was significantly decreased in hypoxic CAFs in HIF-1α-independent manner. Silencing of COL4A2 expression in normoxic CAFs phenocopied the effect of hypoxic CAFs in promoting GC cell migration.
CONCLUSIONS: The reduced expression of COL4A2 in a hypoxic environment might be associated with the tumor-promoting role of hypoxic CAFs in GC.
METHODS: To investigate the underlying mechanism involved in the regulation of gastric cancer (GC) progression by hypoxic CAFs, we performed secretome profiling. Normoxic or hypoxic CAFs conditioned media (CM) were filter-concentrated and in-gel trypsin digested. Resulting peptides were analyzed with LC-MS/MS.
RESULTS: We observed that CM derived from hypoxic CAFs could promote migration of a panel of GC cell lines (AGS, SNU668, SNU638). Mass spectrometry analysis of hypoxic or normoxic CAFs CM identified 1595 proteins, of which 19 proteins (10 upregulated and 9 downregulated) were differentially expressed in the hypoxic secretome. We focused on COL4A2, whose expression was significantly decreased in hypoxic CAFs in HIF-1α-independent manner. Silencing of COL4A2 expression in normoxic CAFs phenocopied the effect of hypoxic CAFs in promoting GC cell migration.
CONCLUSIONS: The reduced expression of COL4A2 in a hypoxic environment might be associated with the tumor-promoting role of hypoxic CAFs in GC.
摘要：
背景：癌症相关成纤维细胞（CAF）是肿瘤微环境（TME）的主要组成部分。已知低氧TME促进肿瘤进展。然而,缺氧条件如何调节CAFs仍然难以捉摸。
方法：为了研究低氧CAFs调节胃癌（GC）进展的潜在机制，我们进行了分泌组分析.将常氧或低氧CAF条件培养基(CM)过滤浓缩并在凝胶内胰蛋白酶消化。用LC-MS/MS分析所得肽。
结果：我们观察到，源自低氧CAFs的CM可以促进一组GC细胞系的迁移（AGS，SNU668，SNU638）。低氧或常氧CAFs的质谱分析CM鉴定出1595种蛋白质，其中19种蛋白质（10种上调，9种下调）在低氧分泌组中差异表达。我们专注于COL4A2，其在缺氧CAFs中以HIF-1α非依赖性方式显着降低。COL4A2在常氧CAFs中表达的沉默证实了低氧CAFs促进GC细胞迁移的作用。
结论：COL4A2在低氧环境中的表达降低可能与低氧CAFs在GC中的促肿瘤作用有关。
方法：为了研究低氧CAFs调节胃癌（GC）进展的潜在机制，我们进行了分泌组分析.将常氧或低氧CAF条件培养基(CM)过滤浓缩并在凝胶内胰蛋白酶消化。用LC-MS/MS分析所得肽。
结果：我们观察到，源自低氧CAFs的CM可以促进一组GC细胞系的迁移（AGS，SNU668，SNU638）。低氧或常氧CAFs的质谱分析CM鉴定出1595种蛋白质，其中19种蛋白质（10种上调，9种下调）在低氧分泌组中差异表达。我们专注于COL4A2，其在缺氧CAFs中以HIF-1α非依赖性方式显着降低。COL4A2在常氧CAFs中表达的沉默证实了低氧CAFs促进GC细胞迁移的作用。
结论：COL4A2在低氧环境中的表达降低可能与低氧CAFs在GC中的促肿瘤作用有关。
