Department of Gastrointestinal Surgery, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang 150081, People's Republic of China.
Department of Gastrointestinal Surgery, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang 150081, People's Republic of China.
Int Immunopharmacol. 2020 Jan;78:105937. doi: 10.1016/j.intimp.2019.105937. Epub 2019 Nov 19.
As a common disease, gastric cancer (GC) has influenced over 1 million people worldwide. Despite of prevention and optimal treatments, GC still has a high mortality. The role of cancer-associated fibroblasts (CAFs) in tumor progression has recently attracted attention, yet few studies have focused on GC. Estrogen has been reported to relate to the poor prognosis of GC. Therefore, we investigated whether estrogen can stimulate CAFs to produce tumor promoting factors in this study. Gastric CAFs were isolated from GC tissues and treated with estrogen. ELISA results suggested that CAFs produced interleukin-6 (IL-6) after estrogen treatment in a dose-dependent manner. The cell culture supernatant for estrogen-treated CAFs was collected and used as conditioned medium (CM) for GC cells. After cultured in CM, increased cell proliferation and alteration of cell cycle were detected by CCK-8 assay, BrdU assay, and flow cytometry. Western blot and gelatin zymography were used to determine cancer invasion-associated proteins. Results indicated that the expression of matrix metalloproteinase 2 (MMP2) and MMP9 were enhanced by Estrogen-CAFs-CM. Additional transwell assay showed that cell invasion and migration were promoted after cultured in CM. Lastly, western blot and immunofluorescence results demonstrated that the level of phosphorylated signal transducer and activator of transcription 3 (STAT3) in GC cells increased after cultured in CM. The effect was neutralized by IL-6 neutralizing antibody and STAT3 siRNA. Conclusively speaking, estrogen can activate CAFs to produce IL-6, ending up with promotion of GC cell proliferation and invasion. This result may suggest a new therapeutic target for GC.
作为一种常见疾病，胃癌（GC）已经影响了全球超过 100 万人。尽管采取了预防和最佳治疗措施，GC 的死亡率仍然很高。癌症相关成纤维细胞（CAFs）在肿瘤进展中的作用最近引起了关注，但很少有研究关注 GC。据报道，雌激素与 GC 的预后不良有关。因此，我们在这项研究中调查了雌激素是否可以刺激 CAFs 产生促进肿瘤的因子。从 GC 组织中分离出胃 CAFs，并用雌激素处理。ELISA 结果表明，CAFs 在雌激素处理后以剂量依赖性方式产生白细胞介素-6（IL-6）。收集经过雌激素处理的 CAFs 的细胞培养上清液，并用作 GC 细胞的条件培养基（CM）。在 CM 中培养后，通过 CCK-8 测定、BrdU 测定和流式细胞术检测到细胞增殖增加和细胞周期改变。Western blot 和明胶酶谱法用于确定与癌症侵袭相关的蛋白质。结果表明，基质金属蛋白酶 2（MMP2）和 MMP9 的表达通过雌激素-CAFs-CM 增强。另外的 Transwell 测定表明，在 CM 中培养后细胞侵袭和迁移能力增强。最后，Western blot 和免疫荧光结果表明，GC 细胞中磷酸化信号转导和转录激活因子 3（STAT3）的水平在 CM 中培养后增加。该作用可被 IL-6 中和抗体和 STAT3 siRNA 中和。总之，雌激素可以激活 CAFs 产生 IL-6，最终促进 GC 细胞的增殖和侵袭。这一结果可能为 GC 提供了一个新的治疗靶点。
