The Laboratory for Tumor Molecular Diagnosis, Affiliated People's Hospital of Inner Mongolia Medical University, Hohhot, 010020, China.
Department of Gastroenterology, Affiliated People's Hospital of Inner Mongolia Medical University, Hohhot, 010020, China.
Chin J Integr Med. 2019 Nov;25(11):837-844. doi: 10.1007/s11655-018-2985-3. Epub 2018 Oct 25.
To investigate the inhibitory effects of paeoniflorin on migration- and invasion-promoting capacities of gastric cancer associated fibroblasts (GCAFs) and to explore the molecular mechanism underlying the effects.
Paired gastric normal fifbroblast (GNF) and GCAF cultures were established from resected tissues. GCAFs were treated with control medium, or 2.5, 5 or 10 μg/mL paeoniflorin. Conditioned media were prepared from GNFs, GCAFs, control-treated GCAFs and paeoniflorin-treated GCAFs, and used to culture AGS human gastric cancer cells. The migration and invasion capacities of AGS cells were determined with wound healing test and transwell invasion assay, respectively. The interleukin 6 (IL-6) mRNA and microRNA-149 expression in GCAFs were detected by reverse transcription-quantitative polymerase chain reaction. The IL-6 protein expression and secretion by GCAFs were measured with Western blot and enzyme-linked immunosorbent assay analysis, respectively. The protein levels of phosphorylated signal transducer and activator of transcription 3 (STAT3), matrix metalloproteinase (MMP) and MMP9 in AGS cells were examined by Western blot.
GCAFs displayed enhanced capacities to induce AGS cell migration and invasion as compared with GNFs. Paeoniflorin treatment significantly inhibited the migration- and invasion-promoting capacities of GCAFs (P<0.05). GCAFs produced and secreted more IL-6 into the conditioned medium than GNFs, leading to over-activation of STAT3-MMP signaling in AGS cells. Paeoniflorin suppressed IL-6 production and secretion by up-regulating microRNA149 expression in GCAFs, and subsequently prevented GCAFs from activating IL-6-STAT3-MMP signaling of AGS cells.
Paeoniflorin inhibits the migration- and invasion-promoting capacities of GCAFs by targeting microRNA-149 and IL-6. Paeoniflorin is potentially a novel therapeutic agent against cancer microenvironment.
研究芍药苷对胃癌相关成纤维细胞（GCAFs）迁移和侵袭能力的抑制作用，并探讨其作用机制。
从切除的组织中建立配对的胃正常成纤维细胞（GNF）和 GCAF 培养物。用对照培养基或 2.5、5 或 10μg/ml 芍药苷处理 GCAFs。从 GNFs、GCAFs、对照处理的 GCAFs 和芍药苷处理的 GCAFs 中制备条件培养基，并用于培养 AGS 人胃癌细胞。通过划痕愈合试验和 Transwell 侵袭试验分别测定 AGS 细胞的迁移和侵袭能力。用逆转录定量聚合酶链反应检测 GCAFs 中的白细胞介素 6（IL-6）mRNA 和 microRNA-149 的表达。用 Western blot 和酶联免疫吸附试验分别检测 GCAFs 中 IL-6 蛋白的表达和分泌。用 Western blot 检测 AGS 细胞中磷酸化信号转导和转录激活因子 3（STAT3）、基质金属蛋白酶（MMP）和 MMP9 的蛋白水平。
与 GNFs 相比，GCAFs 显示出增强的诱导 AGS 细胞迁移和侵袭的能力。芍药苷处理显著抑制 GCAFs 的迁移和侵袭促进能力（P<0.05）。GCAFs 比 GNFs 产生和分泌更多的 IL-6 进入条件培养基，导致 AGS 细胞中 STAT3-MMP 信号过度激活。芍药苷通过上调 GCAFs 中 microRNA149 的表达抑制 IL-6 的产生和分泌，从而防止 GCAFs 激活 AGS 细胞中的 IL-6-STAT3-MMP 信号。
芍药苷通过靶向 microRNA-149 和 IL-6 抑制 GCAFs 的迁移和侵袭促进能力。芍药苷可能是一种针对癌症微环境的新型治疗剂。
