摘要
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 observe the effect of Jiedu Sangen Decoction （JSD, consisting of Polygonum cuspidatum, Geum Japonicum Thumnb, Radix Actinidiae Chinensis） and resveratrol on the metastasis capability of colon cancer CT-26 cells, and on the Sirtl-NF-κB signal pathway of Cancer-associated fibroblasts （CAFs）. Methods： The BALB/C mice were randomly divided into normal control group, blank group, model group, resveratrol group and Jiedu Sangen Decoction group. The serum was collected after modeling. CT-26 supernatant was collected, was used to make conditioned medium with fresh medium by the ratio of 1 ： 1. Mouse fibroblast L929 cells at ogarithmic phase were collected, and incubated with conditioned medium for 72 hours to establish CAFs model. The effects of four different serums on the migration ability of colon cancer CT-26 cells were observed using Transwell. The expression quantities of Sirtl and NF-κB pathway after intervention in normal mice were detected by Western blot. The expression quantities and acting position of Sirtl and NF-κB pathway in CFAs were detected by immunofluorescence assay. Results： The number of CT-26 cells in the JSD treated group and resveratrol group significantly decreased, when compared with the model group, showing statistical significance （P〈0.01）. Compared with other groups, the expressions of Sirtl of CAFs increased in the JSD treated group and resveratrol group at 24h, the expressions of phosphorylation P65 significantly decreased, and the expressions of phosphorylation IKBκ with phosphorylation IKK α β increased significantly, showing statistical significance （P〈0.05）. Compared with other groups, the Sirtl fluorescence significantly enhanced in JSD groups and resveratrol group, and interaction with NF-κB in the nucleus. Conclusion： The JSD and resveratrol can inhibit metastasis ability of colon cancer CT-26 ceils, enhance the expression of Sirtl of CAFs, and inhibit the activation of NF-κB pathway of CAFs.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2015年第9期3082-3086,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81373632)
浙江省中医药优秀青年人才基金计划(No.2013ZQ011)
浙江省教育厅项目(No.Y201016272)~~
关键词
虎杖根
水杨梅根
藤梨根
白藜芦醇
结肠癌
侵袭转移
Polygonum cuspidatum
Geum Japonicum Thumnb
Radix Actinidiae Chinensis
Resveratrol
Colorectal cancer
Invasion and metastasis
