黄芪提取物黄芪多糖通过miR-25/FBXW7信号通路抑制胃癌细胞增殖和侵袭  

Astragalus Polysaccharides Inhibiting Gastric Cancer Cell Proliferation and Invasion via miR-25/FBXW7 Signaling Pathway

作　　者：李萌 赵伟 耿丽媛 姜静 杨楠 谷莉莉 王利平 王毓麟 LI Meng;ZHAO Wei;GENG Liyuan;JIANG Jing;YANG Nan;GU Lili;WANG Liping;WANG Yulin(Department of Gastroenterology,Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital,Tianjin 300120,China;Digestive endoscopy center,Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital,Tianjin 300120,China)

机构地区：[1]天津市中医药研究院附属医院消化科,天津300120 [2]天津市中医药研究院附属医院内镜中心,天津300120

出　　处：《新中医》2025年第3期150-155,共6页New Chinese Medicine

摘　　要：目的:观察黄芪提取物黄芪多糖(APS)对胃癌(GC)细胞系HS-746T细胞的作用及可能机制。方法:通过TargetScan工具预测microRNA-25 (miR-25)与FBXW7结合情况,双荧光素酶报告实验测定miR-25和FBXW7之间的结合关系。体外培养人胃癌细胞系HS-746T细胞,按实验需求将细胞分为control组(转染空白对照基因)、inhibitor组(转染miR-25抑制基因)、APS组(转染空白对照基因,加入1mg/mLAPS的DMEM培养基培养24 h)、miR-25 mimics组(转染miR-25过表达基因)、APS+miR-25 mimics组(转染miR-25过表达基因,加入1mg/mLAPS的DMEM培养基培养24h)。实时荧光定量PCR检测各组miR-25mRNA和FBXW7 mRNA的表达水平,MTT实验检测各组细胞增殖情况,Transwell实验检测各组细胞的侵袭能力。结果:Starbase工具预测miR-25与FBXW7结合,双荧光素酶报告实验测定揭示了miR-25和FBXW7之间可直接结合。与control组比较,inhibitor组miR-25 mRNA表达水平下降,FBXW7 mRNA表达水平升高(P<0.05);APS组miR-25 mRNA的表达降低,FBXW7 mRNA的表达增多(P<0.05);miR-25 mimics组的miR-25 mRNA表达水平升高,FBXW7 mRNA表达水平降低(P<0.05)。与miR-25 mimics组比较,APS+miR-25 mimics组miR-25mRNA的表达明显减少,FBXW7 mRNA的表达明显增加(P<0.05)。MTT实验结果显示,实验48 h,inhibitor组细胞增殖率低于control组(P<0.05),miR-25 mimics组细胞增殖率高于control组(P<0.05)。Transwell实验结果显示,inhibitor组穿膜细胞数低于control组(P<0.05),miR-25mimics组穿膜细胞数高于control组(P<0.05)。结论:APS可以抑制HS-746T细胞的增殖和侵袭,抑制miR-25 mRNA的表达,促进FBXW7mRNA的表达,也可以逆转细胞转染miR-25 mimic引起的miR-25升高和FBXW7抑制。APS可能通过miR-25/FBXW7信号通路抑制GC细胞HS-746T的增殖和侵袭。Objective:To observe the effects and potential mechanisms of Astragalus polysaccharides(APS)on the gastric cancer(GC)cell line HS-746T.Methods:The binding of microRNA-25(miR-25)to FBXW7 was predicted using the TargetScan tool,and the relationship between miR-25 and FBXW7 was determined using a dual-luciferase reporter assay.Human gastric cancer cell line HS-746T cells were cultured in vitro and divided into the following groups based on experimental needs:the control group(transfected with blank control gene),inhibitor group(transfected with miR-25 inhibitor),APS group(transfected with blank control gene and cultured in DMEM medium containing 1 mg/mL APS for 24 hours),miR-25 mimics group(transfected with miR-25 overexpression gene),and APS+miR-25 mimics group(transfected with miR-25 overexpression gene and cultured in DMEM medium containing 1 mg/mL APS for 24 hours).The expression levels of miR-25 mRNA and FBXW7 mRNA in each group were detected by real-time quantitative PCR;cell proliferation was measured by MTT assay,and cell invasion ability was assessed by Transwell assay.Results:The Starbase tool predicted the binding of miR-25 to FBXW7,and the dual-luciferase reporter assay revealed a direct binding relationship between miR-25 and FBXW7.Compared with those in the control group,the miR-25 mRNA expression level decreased and the FBXW7 mRNA expression level increased in the inhibitor group(P<0.05);miR-25 mRNA expression decreased and FBXW7 mRNA expression increased in the APS group(P<0.05);miR-25 mRNA expression increased and FBXW7 mRNA expression decreased in the miR-25 mimics group(P<0.05).Compared with those in the miR-25 mimics group,miR-25 mRNA expression significantly decreased and FBXW7 mRNA expression significantly increased in the APS+miR-25 mimics group(P<0.05).MTT assay results showed that after 48 hours,the cell proliferation rate in the inhibitor group was lower than that in the control group(P<0.05),and the cell proliferation rate in the miR-25 mimics group was higher than that in the control grou

关 键 词：胃癌 黄芪多糖 微小RNA25 HS-746T FBXW7 

分 类 号：R573[医药卫生—消化系统]