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目的:探讨解毒三根汤(虎杖根、水杨梅根、藤梨根)和白藜芦醇对结肠癌CT-26细胞的侵袭能力的影响及肿瘤相关成纤维细胞(CAFs)的Sirt1-NF-κB信号通路的影响。方法:将BALB/C小鼠随机分成正常对照组、空白组、模型组、白藜芦醇组和解毒三根汤组。造模成功后取血分离血清。收集CT-26上清液,并以1∶1比例与新鲜培养基配置条件培养基;收集对数生长期的小鼠成纤维L929细胞,用条件培养基培养72h建立CAFs模型;Transwell观察4组血清对CT-26细胞迁移力的影响,Western blot检测5组血清干预正常小鼠成纤维细胞后,NF-κB通路和Sirt1表达量,免疫荧光分析CAFs中NF-κB通路和Sirt1表达量及作用位置。结果:解毒三根汤组和白藜芦醇组透膜结肠癌CT-26细胞数量与模型组相比明显减少(P<0.01);24h解毒三根汤组和白藜芦醇组干预的CAFs中Sirt1的表达量较其他组明显上升,磷酸化P65表达显著减少,磷酸化IKBα、磷酸化IKKαβ表达显著增加(P<0.05);解毒三根汤组和白藜芦醇组较其他组Sirt1荧光明显增强,并且与NF-κB共同作用于细胞核中。结论:解毒三根汤和白藜芦醇能明显抑制结肠癌CT-26细胞的侵袭运动,增强CAFs中Sirt1的表达量,抑制CAFs中NF-κB通路的激活。
Objective: To investigate the effect of Jiedu Sangen Tang (Polygonum cuspidatum, Myricetoma rhizome, ramie root) and resveratrol on invasiveness of colon cancer CT-26 cells and the expression of Sirt1-NF-κB in tumor-associated fibroblasts Effects of signal pathways. Methods: BALB / C mice were randomly divided into normal control group, blank group, model group, resveratrol group and detoxification three root group. Separation of blood from the model after successful blood separation. The supernatant of CT-26 was collected and the conditioned medium was placed in a 1: 1 ratio with fresh medium. The mouse fibroblast L929 cells in logarithmic growth phase were collected and cultured in conditioned media for 72 hours to establish CAFs model. Transwell group 4 The effect of serum on the migration of CT-26 cells was detected by Western blot. The expression of NF-κB pathway and Sirt1 in normal mouse fibroblasts were detected by Western blot. The expression of NF-κB pathway and Sirt1 in CAFs were detected by immunofluorescence position. Results: The numbers of CT-26 cells in the Jiedu Sangen decoction group and the resveratrol group were significantly decreased compared with the model group (P <0.01). Among the CAFs treated with the Jiedu Sangen decoction group and the resveratrol group, Sirt1 (P <0.05). Compared with the other groups, the expression of P65 was significantly decreased, the expression of phosphorylated P65 and phosphorylated IKKαβ were significantly increased (P <0.05) And co-acts with NF-κB in the nucleus. Conclusion: Jiedu Sangen Decoction and resveratrol can significantly inhibit colon cancer CT-26 cell invasion motility, enhance the expression of Sirt1 in CAFs, and inhibit the activation of NF-κB pathway in CAFs.