目的 探讨四君子汤能否通过上调 miR-205-5p 表达进而抑制胃癌细胞的迁移力、侵袭力以及上皮间质 转化（EMT）进程，并筛查可能参与上述过程的 miR-205-5p 下游靶基因。方法 四君子汤含药血清制备：SD 大鼠以四君子汤（6.9 g·kg-1 ）灌胃，每日 1 次，共 7 d。（1）将胃癌细胞 MGC-803、AGS 作为研究对象，分组及干 预：生理盐水组（10% 空白血清），四君子汤组（10% 四君子汤含药血清），四君子汤+inhibitor NC 组（10% 四君 子汤含药血清+转染抑制剂阴性对照），四君子汤+ miR-205-5p inhibitor 组（10% 四君子汤含药血清+转染 miR-205-5p 抑制剂），各组血清处理均为 24 h。采用 qPCR 或 Western Blot 法检测各组细胞中 miR-205-5p 表 达水平，Snail、MMP9、TIMP1 mRNA 及蛋白表达水平；划痕实验检测细胞迁移能力；Transwell 实验检测细胞 侵袭能力。（2）以人胃癌 MGC-803 细胞为研究对象，分组及干预：阴性对照组（转染阴性对照小干扰 RNA）， miR-205-5p 模拟物组（转染 miR-205-5p 模拟物）。通过转录组测序分析 miR-205-5p 模拟物组 vs 阴性对照组 的差异表达基因；采用 qPCR 技术对 miR-205-5p 过表达后下调的 3 个基因（ENC1、FGF7、CSF1）的表达水平 进行验证；对差异表达基因进行 GO 功能及 KEGG 通路富集分析。结果 （1）与生理盐水组比较，四君子汤组 MGC-803、AGS 细胞的 miR-205-5p 表达水平均显著升高（P＜0.01）。与四君子汤+ inhibitor NC 组比较，四君 子汤+ miR-205-5p inhibitor 组 MGC-803、AGS 细胞的 miR-205-5p 表达均显著下调（P＜0.01）。（2）与生理盐水 组比较，四君子汤组 MGC-803、AGS 细胞的愈合率明显降低（P＜0.05），穿过基质胶的细胞数目显著减少（P＜ 0.01），细胞 MMP9、Snail mRNA 及蛋白表达显著下调（P＜0.05，P＜0.01），TIMP1 mRNA 及蛋白表达显著上 调（P＜0.05，P＜0.01）。与四君子汤+ inhibitor NC 组比较，四君子汤+ miR-205-5p inhibitor 组 MGC-803、 AGS 细胞的划痕愈合率均明显升高（P＜0.05，P＜0.01），穿过基质胶的细胞数目显著增多（P＜0.01），细胞 MMP9、Snail mRNA 及蛋白表达显著上调（P＜0.05，P＜0.01），TIMP1 mRNA 及蛋白表达显著下调（P＜0.05， P＜0.01）（3）与阴性对照组比较，miR-205-5p 模拟物组 MGC-803 细胞中 miR-205-5p 表达显著上调（P＜ 0.01）。转录组高通量测序结果：与阴性对照组比较，miR-205-5p 模拟物组在过表达 miR-205-5p 的 MGC-803 细胞中共鉴定出 401 个发生显著变化的差异表达基因，其中 195 个基因表达上调，206 个基因表达下调。与阴 性对照组比较，miR-205-5p 模拟物组 MGC-803 细胞在过表达 miR-205-5p 后，ENC1、FGF7、CSF1 mRNA 表 达均显著下调（P＜0.01），与测序得到的差异基因表达结果一致。差异表达基因主要富集在细胞黏附和细胞迁 移等生物学过程，以及癌症的转录失调、PI3K-Akt 信号通路、Rap1 信号通路等方面。结论 miR-205-5p 能 够介导四君子汤对胃癌细胞迁移、侵袭和 EMT 进程的抑制作用，其机制可能与 miR-205-5p 抑制其下游 FGF7、ENC1、CSF1 等基因表达有关。

Objective To investigate whether Sijunzi Decoction （SJZT） inhibits the migration，invasion and epithelial- mesenchymal transition （EMT） process of gastric cancer cells by up-regulating the expression of miR-205-5p，and to screen the downstream target genes of miR-205-5p that might be involved in the above process. Methods Preparation of serum containing SJZT：SD rats were given SJZT （6.9 g·kg-1 ） by gavage once a day for 7 days. （1） Gastric cancer cells MGC-803 and AGS were used as research objects. Grouping and intervention：normal saline group（10% blank serum） ，SJZT group（10% SJZT containing serum） ，SJZT + inhibitor NC group（10% SJZT containing serum + transfection inhibitor negative control）， SJZT + miR-205-5p inhibitor group （10% SJZT containing serum + transfection miR-205-5p inhibitor） . The serum treatment of each group was 24 hours. The mRNA and protein expression levels of miR-205-5p，Snail，MMP9 and TIMP1 in each group were detected by qPCR and Western Blot. Scratch assay was used to detect cell migration ability；transwell assay was used to detect cell invasion ability. （2） Human gastric cancer MGC-803 cells were used as the research object，grouping and intervention：negative control group （transfected with negative control small interfering RNA），miR-205-5p mimic group （transfected with miR-205-5p mimic） . The differentially expressed genes of miR-205-5p mimic group versus negative control group were analyzed by transcriptome sequencing. The expression levels of three genes（ENC1，FGF7，CSF1） down-regulated by miR-205-5p overexpression were verified by qPCR. GO function and KEGG pathway enrichment analysis were performed on differentially expressed genes. Results（1）Compared with the normal saline group，the expression levels of miR-205-5p in MGC-803 and AGS cells in the SJZT group were significantly increased（P＜0.01） . Compared with SJZT + inhibitor NC group，the expression of miR-205-5p in MGC-803 and AGS cells in SJZT + miR-205-5p inhibitor group was significantly down-regulated （P＜0.01） . （2） Compared with the normal saline group，the healing rate of MGC-803 and AGS cells in the SJZT group was significantly decreased （P＜0.05），the number of cells passing through the matrigel was significantly decreased （P＜0.01）， the mRNA and protein expressions of MMP9 and Snail were significantly down-regulated （P＜0.05， P＜0.01）， and the mRNA and protein expressions of TIMP1 were significantly up-regulated（P＜0.05，P＜0.01） . Compared with SJZT + inhibitor NC group，the scratch healing rate of MGC-803 and AGS cells in SJZT + miR-205-5p inhibitor group was significantly increased （P＜0.05，P＜0.01）， the number of cells passing through matrigel was significantly increased（P＜0.01） ，the mRNA and protein expressions of MMP9 and Snail were significantly up-regulated （P＜0.05，P＜0.01） ，and the mRNA and protein expressions of TIMP1 were significantly down-regulated （P＜0.05，P＜0.01） .（3） Compared with the negative control group. The expression of miR-205-5p in MGC-803 cells was significantly up-regulated in miR-205-5p mimics group（P＜0.01） . Transcriptome high-throughput sequencing results： compared with the negative control group， a total of 401 differentially expressed genes with significant changes were identified in MGC-803 cells overexpressing miR-205-5p in the miR-205-5p mimic group，of which 195 genes were up-regulated and 206 genes were down-regulated. Compared with the negative control group，the mRNA expressions of ENC1，FGF7 and CSF1 in MGC-803 cells of miR-205-5p mimic group was significantly down-regulated after overexpression of miR-205-5p （P＜0.01），which was consistent with the results of differential gene expression obtained by sequencing. The differentially expressed genes were mainly enriched in biological processes such as cell adhesion and cell migration，as well as transcription disorders of cancer， PI3K-Akt signaling pathway，Rap1 signaling pathway and so on. Conclusion miR-205-5p can mediate the inhibitory effect of SJZT on the migration，invasion and epithelial-mesenchymal transition of gastric cancer cells. The mechanism may be related to the inhibition of miR-205-5p on the expression of downstream genes such as FGF7，ENC1 and CSF1.

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国家自然科学基金项目（81803855）；辽宁省科技计划联合计划应用基础研究项目（2023JH2/101700205）；辽宁省“兴辽英才计划”项目 （XLYC2203180）；辽宁省教育厅科技创新团队项目（LJ222410162020）