调脾安肠方对裸鼠结肠癌肝转移灶miR-34a-5p、Notch1/NF-KB表达的影响

扫码查看

原文链接

万方数据

中文摘要：目的探讨调脾安肠方对裸鼠结肠癌肝转移(colorectal cancer liver metastasis,CLM)灶组织中 MicroRNA-34a-5p(miR-34a-5p)及其靶基因 Notch1/核转录因子-κB(nuclear factor-kappa B,NF-κB)通路表达的影响.方法选取30只雄性Balb/c裸鼠随机分为空白组、模型组、中药组,每组10只.脾脏原位注射HCT-116人结肠癌细胞(细胞数约为1×107个/mL)法构建CLM模型.中药组在CLM造模完成2周后开始调脾安肠方(2 g/mL,每日0.2 mL)灌胃2周,空白组和模型组同期予等体积0.9％生理盐水灌胃.苏木素—伊红染色观察肝转移灶病理组织形态;蛋白免疫印迹法测定肝转移灶Notch1、NF-κB p65蛋白水平;实时荧光定量PCR测定肝转移灶miR-34a-5p及Notch1、NF-κB p65 mRNA水平.结果与模型组比较,中药组裸鼠镜下病理转移灶面积明显减小,核分裂像减少,提示调脾安肠方抑制了结直肠癌细胞在肝脏的定植和迁移.与空白组比较,模型组肝转移灶中 miR-34a-5p 含量显著降低(P＜0.01),Notch1、NF-κB p65 蛋白表达升高(均 P＜0.05),Notch1、NF-κB p65的mRNA表达显著升高(均P＜0.01).与模型组比较,中药组Notch1蛋白及mRNA表达降低(P＜0.05),NF-κB p65 的 mRNA 表达显著降低(P＜0.01).miR-34a-5p 与 Notch1 mRNA 在结肠癌肝转移灶组织中表达水平呈负相关(r2=0.8845,P＜0.05),Notch1与NF-κB p65在结肠癌肝转移灶组织中mRNA表达水平呈显著正相关(r2=0.9291,P＜0.01).结论中药复方调脾安肠方能抑制裸鼠结肠癌肝转移灶形成,其机制可能与通过抑癌基因miR-34a-5p靶向负性调节Notch1/NF-KB通路表达有关.

外文标题：Effect of Tiaopi Anchang Decoction on the expression of miR-34a-5p and Notch1/NF-κB in nude mice with liver metastasis of colorectal cancer

外文摘要：Objective To investigate the effect of Tiaopi Anchang decoction on the expression of miR-34a-5p and its target gene Notch1/NF-KB pathway in liver metastases of colorectal cancer(CLM)in nude mice.Methods Thirty male Balb/C nude mice were randomly divided into control group,model group and drug administration group,with 10 mice in each group.The CLM model was constructed by injecting HCT-116 human colorectal cancer cells into the spleen in situ.In the administration group,Tiaopi Anchang Decoction was administered by gavage after modeling,and HE staining was used to observe the pathological morphology of liver metastases.The protein levels of Notch1 and NF-κB p65 in liver metastases were determined by Western blot.The mRNA levels of miR-34a-5p,Notch1 and NF-κB p65 in liver metastases were determined by RT-PCR.Results Compared with the model group,the area of pathological metastases and mitotic figures of nude mice in the administration group were significantly decreased,and the central necrotic area was formed.Tiaopi Anchang Decoction inhibited the colonization and migration of CRC cells in the liver.Compared with the control group,the content of miR-34a-5p in liver metastases in the model group was significantly decreased(P＜0.01),the protein expressions of Notch1 and NF-κB p65 were increased(P＜0.05,P＜0.05),the mRNA expressions of Notch1 and NF-κB p65 were significantly increased(P＜0.01,P＜0.01);compared with the model group,the expression of Notch1 protein and mRNA was decreased in the administration group(P＜0.05),the mRNA expression of NF-κB p65 was significantly decreased(P＜0.01);the expression levels of miR-34a-5p and Notch1 mRNA in CLM were negatively correlated(r2=0.8845,P＜0.05);the mRNA expression levels of Notch1 and NF-κB p65 in liver metastases of colorectal cancer were significantly positively correlated(r2=0.9291,P＜0.01).Conclusion Tiaopi Anchang decoction may negatively regulate the expression of Notch1/NF-KB pathway by targeting miR-34a-5p,and inhibit the formation of CLM in nude mice.

外文关键词：

liver metastases from colon cancertraditional Chinese medicine compoundTiaopi Anchang decoctionmiR-34aNotch1NF-κB p65pharmacological mechanism

作者：

王玉坤、孙适然、张津铖、程志强

展开 >

作者单位：

100029 北京中医药大学中日友好医院临床医学院

中日友好医院中西医结合肿瘤内科

关键词：

结肠癌肝转移中药复方调脾安肠方 miR-34a Notch1 核转录因子-κB p65 药理机制

基金：

北京市自然科学基金中日友好医院高水平医院临床业务费专项临床研究项目

项目编号：

72121942022-NHLHCRF-LX-02-0109

出版年：

2024

DOI：

10.3969/j.issn.1674-1749.2024.03.005

环球中医药

中华国际医学交流基金会

环球中医药

CSTPCD

影响因子：1.553

ISSN：1674-1749

年,卷(期)：2024.17(3)

参考文献量32