[1]李瑞忠,林雁鸿,梁超华,等.黄芪多糖调控SP1/Wnt/β-catenin信号轴抑制骨肉瘤增殖和转移能力的作用机制研究[J].现代检验医学杂志,2025,40(03):37-41,46.[doi:10.3969/j.issn.1671-7414.2025.03.007]
　LI Ruizhong,LIN Yanhong,LIANG Chaohua,et al.Mechanism of the Inhibitory Effect of Astragalus Polysaccharide on the Proliferation and Metastasis of Osteosarcoma Regulate by SP1/Wnt/β-catenin Signaling Axis[J].Journal of Modern Laboratory Medicine,2025,40(03):37-41,46.[doi:10.3969/j.issn.1671-7414.2025.03.007]
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黄芪多糖调控SP1/Wnt/β-catenin信号轴抑制骨肉瘤增殖和转移能力的作用机制研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年03期
页码:
37-41,46
栏目:
论著
出版日期:
2025-05-15

文章信息/Info

Title:
Mechanism of the Inhibitory Effect of Astragalus Polysaccharide on the Proliferation and Metastasis of Osteosarcoma Regulate by SP1/Wnt/β-catenin Signaling Axis
文章编号:
1671-7414（2025）03-037-06
作者:
李瑞忠1，林雁鸿1，梁超华1，马鑫雨1，赵省省1，梁道臣2
（1.广东医科大学第一临床医学院，广东湛江 524001；2.广东医科大学中山市人民医院骨科，广东中山 528400）
Author(s):
LI Ruizhong1，LIN Yanhong1，LIANG Chaohua1，MA Xinyu1，ZHAO Shengsheng1，LIANG Daochen2
(1.the First Clinical Medical College of Guangdong Medical University，Guangdong Zhanjiang 524001，China； 2. Department of Orthopedics，Zhongshan People’s Hospital of Guangdong Medical University，Guangdong Zhongshan 528400，China)
关键词:
骨肉瘤; 黄芪多糖; 细胞增殖; 细胞转移; SP1/Wnt/β-catenin信号轴
分类号:
R738.1；R730.5
DOI:
10.3969/j.issn.1671-7414.2025.03.007
文献标志码:
A
摘要:
目的研究黄芪多糖（APS）抑制骨肉瘤细胞增殖和转移能力的作用机制。方法细胞计数实验（CCK8）检测APS抑制骨肉瘤细胞U2OS,HOS,MG63和成骨细胞hFOB1.19的增殖能力，选择APS对骨肉瘤细胞抑制作用最显著的细胞株进行后续实验；将骨肉瘤细胞分为对照组（NC组）、APS组、APS与转染SP1敲减质粒共处理组（APS+sh-SP1）和APS与转染SP1过表达质粒共处理组（APS+oe-SP1组），蛋白质免疫印迹（WesternBlotting）法检测各组细胞SP1蛋白的表达水平；CCK8实验检测各组细胞增殖能力。Transwell实验检测各组细胞转移能力；TOP/FOPFlash实验检测各组细胞Wnt/β-catenin信号通路活性；WesternBlotting法检测各组细胞Wnt3a，β-catenin，细胞周期蛋白（CyclinD1）、cMYC，基质金属蛋白酶2（MMP2）和Snail蛋白表达。结果骨肉瘤细胞U2OS，HOS，MG63和成骨细胞hFOB1.19经APS处理48h后，细胞增殖抑制率分别为62.93%±4.79%，20.66%±1.10%，39.31%±3.20%和5.97%±0.72%，与成骨细胞hFOB1.19相比，APS显著抑制骨肉瘤细胞的增殖能力，差异具有统计学意义（F=208.400，P＜0.001），且对骨肉瘤细胞U2OS的抑制作用最显著（t=20.380，P＜0.001）。与NC组相比，APS组SP1蛋白表达、细胞增殖能力、穿膜细胞数、细胞中Wnt/β-catenin信号通路活性及Wnt/β-catenin信号通路关键蛋白Wnt3a，β-catenin，下游增殖相关蛋白CyclinD1，cMYC，下游转移相关蛋白MMP2，Snail表达均降低，差异具有统计学意义（t=9.740～90.780，均P＜0.05）。与APS组相比，APS+sh-SP1组细胞中SP1蛋白表达、细胞增殖能力、穿膜细胞数、细胞中Wnt/β-catenin信号通路活性及Wnt/β-catenin信号通路相关蛋白表达进一步降低，差异具有统计学意义（t=3.032～12.940，均P＜0.05）；而APS+oe-SP1组细胞中SP1蛋白表达、细胞增殖能力、穿膜细胞数、细胞中Wnt/β-catenin信号通路活性及细胞中Wnt/β-catenin信号通路相关蛋白表达增加，差异具有统计学意义（t=3.350～22.450，均P＜0.05）。结论APS靶向负调控SP1/Wnt/β-catenin信号轴抑制骨肉瘤细胞增殖和转移能力。
Abstract:
Objective To investigate the mechanism of astragalus polysaccharide (APS) in inhibiting the proliferation and metastasis of osteosarcoma cells. Methods Cell counting kit-8(CCK8) assay was used to detect the inhibitory effect of APS on the proliferation of osteosarcoma cell lines U2OS, HOS, MG63 and osteoblast cell line hFOB1.19，the cell line with the most significant inhibitory effect of APS on osteosarcoma cells was selected for subsequent experiments. Osteosarcoma cells were divided into control group (NC group), APS group, APS+sh-SP1 co treatment group with transfected SP1 knockdown plasmid (APS+sh-SP1), and APS+oe SP1 co treatment group with transfected SP1 overexpression plasmid (APS+oe SP1 group). Western blotting was used to detect the expression of SP1 protein in each group．CCK8 assay was used to detect the proliferation ability of each group．Transwell assay was used to detect the metastatic ability of cells in each group．TOP/FOP Flash assay was used to detect the activity of Wnt/β-catenin signaling pathway．The protein expressions of Wnt3a, β-catenin, CyclinD1, cMYC,matrix metalloproteinase 2(MMP2) and Snail were detected by western blotting. Result After 48h of APS treatment, the cell proliferation inhibition rates of osteosarcoma cells U2OS, HOS, MG63 and osteoblast cells hFOB1.19 were 62.93%±4.79%, 20.66%±1.10%, 39.31%±3.20% and 5.97%±0.72%, respectively. Compared with osteoblast hFOB1.19, APS significantly inhibited the proliferation of osteosarcoma cells，and the difference was statistically significant (F=208.400, P < 0.001), and the inhibitory effect on osteosarcoma U2OS cells was the most significant (t=20.380, P < 0.001).Compared with NC group, SP1 protein expression, cell proliferation ability, number of transmembrane cells，Wnt/β-catenin signaling pathway activity in cells, Wnt/β-catenin signaling pathway key proteins Wnt3a，β-catenin，downstream proliferation-related protein CyclinD1， cMYC，and downstream metastasis related proteins MMP2 and Snail in the APS group were decreased，and the differences were statistically significant (t=9.740～90.780, all P<0.05).Compared with the APS group, the expression of SP1 protein, cell proliferation ability, the number of transmembrane cells, the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+sh-SP1 group were further decreased，and the differences were statistically significant (t=3.032 ～ 12.940，all P < 0.05). Compared with the APS group, the expression of SP1 protein, cell proliferation ability, the number of transmembrane cells, the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+oe-SP1 group were increased，and the differences were statistically significant (t=3.350 ～ 22.450，all P < 0.05).Conclusion APS inhibits the proliferation and metastasis of osteosarcoma cells by targeting SP1/Wnt/β-catenin signaling axis.

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备注/Memo

备注/Memo:
基金项目：广东省医学科研基金（B2023353）。
作者简介：李瑞忠（1998-），男，硕士研究生，住院医师，研究方向：骨肉瘤，E-mail：1459741483@qq.com。
通讯作者：梁道臣（1971-），男，博士后，主任医师，研究方向：骨质疏松、骨肿瘤，E-mail：liangdc@126.com。
更新日期/Last Update: 2025-05-15