Graduate School of Analytical Science and Technology (GRAST), Chungnam National University, Daejeon, 34134, Republic of Korea.
Advanced Analysis and Data Center, Korea Institute of Science and Technology (KIST), Seoul, 02456, Republic of Korea.
J Cancer Res Clin Oncol. 2023 Jul;149(8):4477-4487. doi: 10.1007/s00432-022-04361-y. Epub 2022 Sep 20.
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.
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.
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.
The reduced expression of COL4A2 in a hypoxic environment might be associated with the tumor-promoting role of hypoxic CAFs in GC.
癌症相关成纤维细胞（CAFs）是肿瘤微环境（TME）的主要组成部分。已知缺氧的 TME 会促进肿瘤的进展。然而，缺氧条件如何调节 CAFs 仍然难以捉摸。
为了研究缺氧 CAFs 调节胃癌（GC）进展的潜在机制，我们进行了分泌组谱分析。将常氧或缺氧 CAFs 的条件培养基（CM）进行过滤浓缩和胶内胰蛋白酶消化。用 LC-MS/MS 分析所得肽段。
我们观察到，缺氧 CAFs 的 CM 可促进一系列 GC 细胞系（AGS、SNU668、SNU638）的迁移。对缺氧或常氧 CAFs CM 的质谱分析鉴定出 1595 种蛋白质，其中 19 种蛋白质（10 种上调和 9 种下调）在缺氧分泌组中差异表达。我们专注于 COL4A2，其在 HIF-1α 非依赖性方式下在缺氧 CAFs 中的表达显著降低。在常氧 CAFs 中沉默 COL4A2 的表达可模拟缺氧 CAFs 促进 GC 细胞迁移的作用。
在缺氧环境中 COL4A2 的表达降低可能与缺氧 CAFs 在 GC 中的促肿瘤作用有关。
