摘要
目的探究健脾通络方对结直肠癌细胞侵袭、迁移的影响及机制。方法构建敲减MALAT1基因的HCT116细胞模型。采用细胞划痕实验和Transwell实验检测细胞侵袭、迁移能力。采用蛋白质印迹法检测经健脾通络方干预后的MALAT1正常/敲减表达的HCT116细胞中Wnt3a、β-catenin蛋白水平。采用实时荧光定量PCR法检测MALAT1的表达水平。构建裸鼠肺转移模型,采用小动物活体成像技术监测肿瘤转移情况,并检测转移灶中Wnt3a、β-catenin和MALAT1的表达水平。结果经健脾通络方干预后,HCT116细胞中MALAT1、Wnt3a和β-catenin的表达水平均降低,HCT116细胞迁移能力下降。在裸鼠肺转移模型中,经健脾通络方干预后裸鼠的转移灶减少,转移灶中Wnt3a、β-catenin和MALAT1的表达水平均降低。结论健脾通络方通过下调HCT116细胞中MALAT1的表达,使Wnt/β-catenin信号通路中Wnt3a、β-catenin的蛋白表达水平降低,从而抑制结直肠癌细胞的增殖、侵袭和迁移能力。
Objective An attempt is made in this paper to investigate the effect and mechanism of Jianpi Tongluo Decoction on invasion and migration of colorectal cancer cells.Methods The MALAT1knockdown HCT116 cell model was constructed.Cell scratch test and Transwell test were conducted to detect cell invasion and migration.Western blotting was conducted to detect Wnt3a andβ-catenin protein levels in HCT116 cells with normal/knockdown expression of MALAT1 after treatment with Jianpi Tongluo Decoction.The expression of MALAT1 was detected by real-time fluorescence quantitative PCR.The lung metastasis model of nude mice was established,and the tumor metastasis was monitored by utilizing the small animal in vivo imaging technology.The expressions of Wnt3a,β-catenin,and MALAT1 in metastatic lesions were detected.Results The expressions of MALAT1,Wnt3a,andβ-catenin in HCT116 cells are decreased after treatment with Jianpi Tongluo Decoction,and the migration ability of HCT116 cells is decreased.In the lung metastasis model of nude mice,the metastatic lesions of the nude mice treated with Jianpi Tongluo Decoction are reduced,and the expressions of Wnt3a,β-catenin,and MALAT1 in the metastatic lesions are decreased.Conclusion Jianpi Tongluo Decoction inhibits the proliferation,invasion,and migration of colorectal cancer cells by down-regulating the expression of MALAT1 in HCT116 cells and mediating the reduction of Wnt3a andβ-catenin protein expression in Wnt/β-catenin signaling pathway.
作者
黄人可
任建琳
靖琳
吴杏黎
刘湘君
刘怡
储金砚
HUANG Renke;REN Jianlin;JING Lin;WU Xingli;LIU Xiangjun;LIU Yi;CHU Jinyan(Department of Oncology,Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China)
出处
《国际消化病杂志》
CAS
2023年第1期39-47,共9页
International Journal of Digestive Diseases
基金
国家自然科学基金面上项目(81873279、82174452、81473628)
上海市科委科技支撑“中药新药临床前研究”项目(22S21901000)
上海申康医院发展中心“临床三年行动计划”疑难疾病精准诊治攻关项目(SHDC2020CR2047B)
上海市中医医院中医药传承院级名中医人才培育-院级名中医(WLJH2021ZY-MZY026)。
