【摘要】 目的：探讨湿热证结直肠癌（CRC）患者肠道菌群对小鼠肠癌肝转移及肿瘤相关成纤维细胞（CAF）活化的影响。方法：采取粪菌移植方法观察湿热证患者肠道菌群对小鼠肝转移的影响；免疫组化检测小鼠肝转移灶中Ki67阳性细胞；16S rRNA分析湿热证CRC患者及粪菌移植小鼠肠道菌群结构；免疫荧光和RT-qPCR检测CAF活化情况；Western Blot检测肝转移灶PI3K/AKT信号通路和上皮-间质转化（EMT）。结果：湿热证CRC粪菌移植组（CRCA-FMT）肝脏转移灶数量显著高于无证可辨CRC粪菌移植组（CRCN-FMT）;CRCA-FMT肝转移灶中Ki67阳性细胞明显高于CRCN-FMT;CRCA-FMT中CAF激活更为明显，且与CAF活化相关的分泌因子CXCL1和TGFβ水平也显著升高；PI3K和AKT磷酸化水平显著增加，同时E-cadherin显著减少、Vimentin显著增加。结论：粪菌移植湿热证CRC肠道菌群可能通过激活CAF和PI3K/AKT信号通路诱导EMT过程，从而促进小鼠肠癌肝转移。更多还原

【Abstract】 Objective: To investigate the effects of gut bacteria isolated from colorectal cancer(CRC) patients with dampheat syndrome on liver metastasis and cancer-associated fibroblasts(CAF) activation in colorectal cancer xenografts mice. Methods:Fecal microbiota transplantation(FMT) technique was used to explore the role of microbiota from CRC patients with dampheat syndrome on liver metastases in mice; Immunohistochemistry was applied for detection of Ki67 positive cells in metastatic liver foci; 16S rRNA was conducted to analyze bacteria composition of CRC donors and mice received with donor microbiota;Immunofluorescence(IF) and RT-qPCR were used for detection of CAF activation; Western Blot was applied to determine PI3K/AKT signaling pathway and epithelial-mesenchymal transition(EMT). Results: The liver nodules in the FMT of CRCA group(CRCA-FMT) were more than that in the FMT of CRC patients with no syndrome group(CRCN-FMT). The positive Ki67 level in CRCA-FMT was significantly higher than that in the CRCN-FMT, IF and RT-qPCR results indicated that the proportion of activated CAF was also much higher in the CRCA-FMT, besides, the levels of CXCL1 and TGFβ were significantly higher than those in the CRCN-FMT; Compared with the CRCN-FMT, PI3K/AKT signaling pathway was activated in CRCA-FMT, moreover, E-cadherin was decreased while vimentin was increased. Conclusion: FMT of CRC patients with damp-heat syndrome could accelerate EMT process by inducing CAF activation and PI3K/AKT signaling pathway, thus promoting the liver metastasis of intestinal cancer in mice.更多还原