健脾解毒方通过调控Wnt/β-catenin信号通路抑制幽门螺杆菌诱导的胃癌细胞干性增强的机制研究  

Mechanism research of Jianpi Jiedu Recipe on inducing stemness enhancement of gastric cancer cells by Helicobacter pylori through regulating Wnt/β-catenin signaling pathway

作　　者：刘宁宁[1] 饶希午 张志鹏 谢咚 季青[1] 谈政 孙明瑜[2] LIU Ning-ning;RAO Xi-wu;ZHANG Zhi-peng;XIE Dong;JI Qing;TAN Zheng;SUN Ming-yu(Department of Oncology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Liver Diseases,Ministry of Education Key Laboratory of Liver and Kidney Diseases,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院肿瘤科,上海201203 [2]上海中医药大学曙光医院肝病研究所,教育部肝肾疾病重点实验室,上海201203

出　　处：《中华中医药杂志》2022年第9期5361-5365,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：上海市自然科学基金项目(No.19ZR1458500);上海市三年行动计划课题[No.ZY(2018-2020)-CCCX-2003-02]。

摘　　要：目的:探讨健脾解毒方对幽门螺杆菌(H. pylori)感染诱导的胃癌细胞干性增强的调控作用及可能的机制。方法:采用H. pylori标准株NCTC11637与人胃癌MKN45细胞共培养的方法感染MKN45细胞,分为对照组,H.pylori感染组,健脾解毒方醇提物低、中、高剂量(0.5、1.0、2.5 mg/mL)组,成球实验观察上述细胞成球能力;Western Blot检测H. pylori感染胃癌细胞后干性转录因子OCT-4、CD44、Nanog、SOX-2的表达变化,检测健脾解毒方对H. pylori感染诱导的人胃癌MKN45细胞干性转录相关因子表达的影响;检测H. pylori感染对胃癌细胞核中β-catenin表达的影响及健脾解毒方的调控作用。结果:H. pylori感染可增强胃癌细胞的成球能力(P<0.01),健脾解毒方低、高剂量组可不同程度抑制胃癌细胞的成球能力(P<0.01)。H.pylori感染可显著增加人胃癌MKN45细胞中OCT-4、CD44、Nanog、SOX-2蛋白表达(P<0.01)。健脾解毒方可以下调H. pylori感染诱导的OCT-4、CD44和SOX-2表达(P<0.01),其中对OCT-4和CD44的作用最强,并呈一定的剂量依赖效应。H. pylori感染MKN45细胞12 h后,β-catenin表达显著增加(P<0.01),与MKN45+H. pylori组比较,健脾解毒方醇提物低、高剂量作用后,胞核β-catenin蛋白表达显著下降(P<0.01)。结论:健脾解毒方可抑制H. pylori诱导的胃癌细胞的成球能力,抑制H. pylori诱导的人胃癌细胞干性转录因子OCT-4、CD44和SOX-2表达,调控Wnt/β-catenin信号通路是其机制之一,这可能是其防治胃癌的重要作用靶点。Objective:To investigate the regulatory effects and possible mechanisms of Jianpi Jiedu Recipe on Helicobacter pylori(H. pylori) infection induced enhancement of stemness of gastric cancer cells. Methods: Human gastric cancer MKN45 cells were infected with standard H. pylori NCTC11637 by co-culture, and cells were divided into 5 groups:control guoup, H. pylori group and low-, medium-, high-dose JPJDR group(low, medium and high doses were 0.5, 1.0,2.5 mg/mL respectively). Spheroid formation assay was used to observe the ability of spheroid formation of each group described above. The expression changes of stemness transcription factors OCT-4, CD44, Nanog, SOX-2 in gastric cancer cells infected with H. pylori were detected by Western Blot. To examine the effects of Jianpi Jiedu Recipe on H. pylori infection induced expression of stemness transcription related factors in human gastric cancer MKN45 cells. To examine the effect of H. pylori infection on the expression of β-catenin in the nucleus of gastric cancer cells and the regulatory effect of Jianpi Jiedu Recipe. Results: H. pylori infection could enhance the spherogenic ability of gastric cancer cells(P<0.01), and the low-and high-dose groups of Jianpi Jiedu Recipe can inhibit the spherogenic ability of gastric cancer cells to various degrees(P<0.01). H. pylori infection significantly increased OCT-4, CD44, Nanog, SOX-2 protein expression in human gastric cancer MKN45 cells(P<0.01). Jianpi Jiedu Recipe could down regulate the expression of OCT-4, CD44 and SOX-2 induced by H. pylori infection(P<0.01), which showed the strongest effect on OCT-4 and CD44 and a certain dose-dependent effect, but not on Nanog. The expression of β-catenin in MKN45 cells was increased after 12 h of infection with H. pylori(P<0.01). The expression of β-catenin in the nucleus of MKN45 cells was decreased after low-and high-dose of alcohol extract of Jianpi Jiedu Recipe, and the difference was statistically significant compared with MKN45+H. pylori group(P<0.01). Conclusion: Jianpi Jie

关 键 词：幽门螺杆菌 健脾解毒方 胃癌 干性转录因子 

分 类 号：R285[医药卫生—中药学]