黄芪提取物黄芪多糖通过miR-223/FBXW7信号通路抑制胶质瘤细胞的增殖和侵袭

doi:10.5246/jcps.2025.01.005

摘要/Abstract

摘要：

黄芪提取物黄芪多糖(APS)作为天然提取物, 已被证实可以通过调节miRNA在多种肿瘤的发展过程中起到抑制作用。然而, miRNA (miR)-223在脑胶质瘤细胞中的确切调控机制尚不清楚。本研究旨在探讨黄芪提取物黄芪多糖(APS)miR-223/FBXW7信号通路对脑胶质瘤细胞的抑制作用及可能机制。通过TargetScan工具预测miR-223与FBXW7结合情况, 双荧光素酶报告实验测定miR-223和FBXW7之间的结合关系。比较胶质瘤和癌旁组织miR-223和FBXW7表达情况, 以及预后随访。用APS处理U87细胞, 或miR-223模拟物miR-223的小干扰RNA (siRNA)转染U87细胞, 检测这些处理对细胞增殖和凋亡的影响, 以及FBXW7表达的影响。miR-223与FBXW7结合, 双荧光素酶报告实验测定进一步证实了miR-223和FBXW7之间直接结合。胶质瘤组织中miR-223 mRNA表达高于癌旁组织, FBXW7 mRNA与蛋白含量低于癌旁组织, miR-223与FBXW7 表达负相关(P < 0.05)。APS可以明显抑制U87细胞的增殖和侵袭, 抑制 miR-223的表达, 促进FBXW7的表达(P < 0.05); 细胞转染miR-223 mimic可以逆转APS引起的miR-223抑制和FBXW7升高(P < 0.05)。因此, 本研究表明APS可能通过miR-223/FBXW7信号通路抑制胶质瘤细胞系U87的增殖和侵袭。

关键词: 胶质瘤, 黄芪多糖, 微小RNA-223, FBXW7

Abstract:

Astragalus extract Astragalus polysaccharide (APS), a natural extract, has been demonstrated to exert inhibitory effects on the development of various tumors by modulating microRNA (miRNA). However, the precise regulatory mechanism of miRNA-223 (miR-223) in brain glioma cells remains unclear. This study aimed to investigate the inhibitory effect of Astragalus extract APS on brain glioma cells through the miR-223/FBXW7 signaling pathway and its potential mechanism. Using the TargetScan tool, we predicted the binding between miR-223 and FBXW7 and confirmed this binding relationship through dual luciferase assay. We compared the expression of miR-223 and FBXW7 in glioma and adjacent tissues and followed up the prognosis. U87 cells were subjected to APS treatment or were transfected with small interfering RNA (siRNA) of miR-223 mimics to examine the effects of these treatments on cell proliferation, apoptosis, and FBXW7 expression. Our results demonstrated that miR-223 directly bound to FBXW7, as confirmed by dual luciferase assay. Moreover, the mRNA expression of miR-223 in glioma tissues was higher than that in paracancerous tissues, while the mRNA and protein levels of FBXW7 in glioma tissues were lower than those in paracancerous tissues. In addition, we observed a negative correlation between the expression of miR-223 and FBXW7 (P < 0.05). APS significantly inhibited the proliferation and invasion of U87 cells, suppressed the expression of miR-223, and promoted the expression of FBXW7 (P < 0.05). Transfection of miR-223 mimic into cells reversed the inhibition of miR-223 and the increase of FBXW7 induced by APS (P < 0.05). Therefore, our findings suggested that APS might inhibit the proliferation and invasion of the glioma cell line U87 through the miR-223/FBXW7 signaling pathway.

Key words: Glioma, Astragalus polysaccharide, miRNA-223, FBXW7

Supporting:

张启财, 霍景瑞, 张雨焱, 蒋奎军, 孙中磊. 黄芪提取物黄芪多糖通过miR-223/FBXW7信号通路抑制胶质瘤细胞的增殖和侵袭[J]. 中国药学(英文版), 2025, 34(1): 55-65.

Qicai Zhang, Jingrui Huo, Yuyan Zhang, Kuijun Jiang, Zhonglei Sun. Astragalus extract Astragalus polysaccharide inhibits proliferation and invasion of glioma cells through the miR-223/FBXW7 signaling pathway[J]. Journal of Chinese Pharmaceutical Sciences, 2025, 34(1): 55-65.

图/表 6

Figure 1. miR-223 targeted inhibition of FBXW7 expression. (A) Prediction information of miR-223 and FBXW7 TargetScan; (B) Effect of miR-223-3p on the 3’ UTR downstream of FBXW7 mRNA or the mutated 3’ UTR insertion of FBXW7 mRNA in 293T cells by dual luciferase gene activity assay. a: Compared with other groups, P < 0.05.

Figure 2. Expression of miR-223 and FBXW7 in glioma tissues. (A) mRNA expression level of miR-223; (B) mRNA expression level of FBXW7, and (C) correlation analysis between miR-223 mRNA and FBXW7 mRNA.

Table 1. Clinical data of glioma patients (x ± s).

Figure 3. Overall postoperative survival of glioma patients. (A) The relationship between miR-223 and overall survival after glioma surgery; (B) The relationship between FBXW7 and overall survival after glioma surgery.

Figure 4. miR-223 and FBXW7 mRNA expression levels. (A) mRNA expression level of miR-223, (B) mRNA expression level of FBXW7. a: Compared with the control group, P < 0.05; b: Compared with the APS group and control group, P < 0.05; c: Compared with the miR25 overexpression group, P < 0.05.

Figure 5. (A) MTT added rate of glioma cells; (B) The number of transwell cells in glioma cells. a: Compared with the control group, P < 0.05; b: Compared with the APS group and control group, P < 0.05; c: Compared with the miR25 overexpression group, P < 0.05.

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