摘要
基于Wnt/β-catenin通路探究葛根芩连汤(Gegen Qinlian Decoction,GQD)对结直肠癌的抗肿瘤作用及对胞质分裂的蛋白质调节因子1(protein regulator of cytokinesis 1,PRC1)核易位的影响。采用BALB/c裸鼠皮下接种CT26结直肠癌细胞株制备结直肠肿瘤体内模型,分别按低(L-GQD)、中(M-GQD)及高(H-GQD)剂量连续灌药干预25 d,模型组(Control)作为空白对照,并记录裸鼠皮下肿瘤生长变化,于末次给药后取出肿瘤后观察。苏木精-伊红染色(hematoxylin-eosin staining,HE)、免疫组化(immunohistochemistry,IHC)和原位末端转移酶标记技术(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)检测肿瘤的增殖和凋亡情况,并通过qRT-PCR和蛋白质免疫印迹分析(western blot,WB)检测Wnt/β-catenin信号及其下游因子的表达变化;此外,WB检测GQD对PRC1的磷酸化水平其亚细胞定位的影响。结果表明:相较于Control组,GQD以剂量依赖方式抑制体内结直肠肿瘤的生长并提高其凋亡水平;IHC结果显示GQD下调了增殖相关蛋白细胞周期蛋白D1(Cyclind1)、增殖标志物Ki-67(Ki67)和血小板-内皮细胞黏附分子(CD31)的表达(P<0.05),同时促进了凋亡相关蛋白胱天蛋白酶3(Caspase 3)和胱天蛋白酶9(Caspase 9)的表达(P<0.05);进一步发现GQD可抑制Wnt/β-catenin信号的激活,推测可能与降低PRC1磷酸化水平并增加其核保留比例有关。GQD可显著抑制结直肠肿瘤的增殖并促进其凋亡,GQD还可抑制Wnt/β-catenin信号通路的激活,且可能与介导PRC1的核易位有关。
Based on the Wnt/β-catenin pathway,to investigate the anti-tumor effect of Gegen Qinlian Decoction(GQD)on colorectal cancer and its effect on nuclear translocation of protein regulator of cytokinesis 1(PRC1).BALB/c nude mice were subcutaneously inoculated with the CT26 colorectal cancer cell line and divided into GQD low(L-GQD),medium(M-GQD),and high(H-GQD)dose groups,with the model group serving as the control.The mice were administered the drug daily for 25 days,and the growth of the tumors was recorded.Following the conclusion of the previous administration,the tumor was surgically excised and subjected to subsequent observation.The proliferation and apoptosis of the tumor were assessed using hematoxylin-eosin staining(HE),immunohistochemistry(IHC),and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining techniques,while the Wnt/β-catenin pathway was examined through qRT-PCR and western blot(WB)analysis.Furthermore,alterations in the expression of catenin signals and downstream factors were investigated.In addition,WB was used to detect the effects of GQD on the phosphorylation level of PRC1 and its subcellular localization.Compared with the model group,GQD inhibited the growth and increased the apoptosis level of colorectal tumors in vivo in a dose-dependent manner.IHC results show that GQD down-regulated the expressions of proliferation-related proteins cyclind1,marker of proliferation Ki-67(Ki67),and platelet endothelial cell adhesion molecule-1(CD31)(P<0.05),and promoted the expressions of approbation-related proteins Caspase 3 and Caspase 9(P<0.05).GQD is further found to inhibit the activation of Wnt/β-catenin signaling,which may be related to the reduction of PRC1 phosphorylation level and the alteration of its nuclear retention ratio.This inhibition of Wnt/β-catenin signaling by GQD is closely associated with the suppression of colorectal tumor proliferation and the promotion of apoptosis.Moreover,GQD may involve in mediating nuclear translocation of PRC1.
作者
蔡蓉
王上
肖柳
周燕萍
胡作为
李云海
CAI Rong;WANG Shang;XIAO Liu;ZHOU Yan-ping;HU Zuo-wei;LI Yun-hai(College of Traditional Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430061,China;Affiliated Hospital of Traditional Chinese and Western Medicine of Hubei University of Chinese Medicine,Wuhan 430022,China)
出处
《科学技术与工程》
北大核心
2025年第8期3142-3151,共10页
Science Technology and Engineering
基金
湖北省教育厅科学技术研究项目(Q20232011)
湖北省自然科学基金联合基金(2024AFD308)
湖北省中医药管理局重点学科建设项目([2023]-2号)。
作者简介
第一作者:蔡蓉(1986-),女,汉族,湖北汉川人,博士研究生,讲师。研究方向:中医药防治肿瘤。E-mail:cairong1009@163.com;通信作者:李云海(1971-),男,汉族,江西九江人,教授,博士研究生导师。研究方向:中医经典理论与临床研究。E-mail:155350975@qq.com。
