| 陈皓,郭丽,于晓涛,等.灵芝提取物通过 PBX3/MAPK通路对胶质瘤细胞恶性生物学行为的作用机制研究[J].安徽医药,2025,29(1):28-33. |
| 灵芝提取物通过 PBX3/MAPK通路对胶质瘤细胞恶性生物学行为的作用机制研究 |
| Study on the mechanism ganoderma lucidum extract on malignant biological behavior of glioma cells by PBX3/MAPK axis |
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| DOI：10.3969/j.issn.1009-6469.2025.01.005 |
| 中文关键词: 灵芝属 神经胶质瘤 前 B细胞白血病同源盒基因 3 丝裂原活化蛋白激酶 恶性生物学行为 |
| 英文关键词: Ganoderma Glioma Pre-B-cell leukemia homeobox3 Mitogen-activated protein kinase Malignant biological behavior |
| 基金项目:漯河市 2022年度重大科技创新专项(漯科〔2022〕45号) |
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| 中文摘要: |
| 目的探究灵芝提取物( GLE)是否可通过前 B细胞白血病同源盒基因 3(PBX3)/丝裂原活化蛋白激酶( MAPK)通路影响 U251人胶质瘤细胞恶性生物学行为。方法 2022年 1月至 2023年 1月进行该研究。含不同浓度 GLE培养液( 0、50、100、 200 mg/L GLE)培养 U251细胞 48 h，采用细胞计数试剂盒(CCK-8)法、流式细胞术、平板克隆实验、细胞划痕试验及迁移和侵袭(Transwell)实验来评估细胞的存活率、凋亡情况、集落形成能力、迁移与侵袭特性；实时定量聚合酶链反应( RT-qPCR)检测 PBX3、细胞外信号调节酶(ERK)mRNA表达水平；蛋白质印迹法检测 PBX3、原癌基因 c-RAF(Raf-1)、磷酸化 Raf-1(p-Raf-1)、信号通路细胞外信号调节酶 1/2(ERK1/2)、磷酸化 ERK1/2(p-ERK1/2)蛋白表达情况。结果 0、50、100、200 mg/L GLE下 U251细胞存活率分别为 100%、(86.62±4.26)%、(67.68±3.49)%、(50.84±3.39)%、(40.13±3.25)%，差异有统计学意义( P<0.05)； 0、50、 100、200 mg/L GLE下 U251细胞凋亡率、集落形成数、划痕愈合率、侵袭细胞数、 PBX3、ERK mRNA及 PBX3、p-Raf-1、p-ERK1/2蛋白相对表达水平比较，差异有统计学意义( P<0.05)；随着 GLE浓度的增加， U251细胞存活率、划痕愈合率、 PBX3与 ERK mRNA相对表达水平及 RAS、PBX3、p-Raf-1、p-MEK1/2、p-ERK1/2蛋白相对表达水平均降低，集落形成数及侵袭细胞数均减少，细胞凋亡率升高； GLE作用效果呈剂量性依赖( P<0.05)。结论 GLE可抑制胶质瘤细胞增殖、克隆形成、迁移及侵袭等恶性生物学特性，并诱导其凋亡，其作用机制可能与阻断 PBX3/MAPK通路的激活相关。 |
| 英文摘要: |
| Objective To investigate whether ganoderma lucidum extract (GLE) can affect the malignant biological behavior of U251human glioma cells through the preb-cell leukemia homolobox gene 3 (PBX3)/mitogen-activated protein kinase (MAPK) axis. Meth. ods From January 2022 to January 2023, U251 cells were cultured with GLE medium (0, 50, 100, 200 mg/L GLE) for 48 h, and CCK-8 assay, flow cytometry, plate cloning assay, cell scratch assay and Transwell assay were used to evaluate cell viability, apoptosis, colo-ny-forming ability, migration, and invasion characteristics. The mRNA expression level of PBX3 and extracellular signal regulating en-zyme (ERK) were detected by RT-qPCR. Western blotting was used to detect the protein expressions of PBX3, proto-oncogene c-Raf (RAF-1), phosphorylated Raf-1 (p-Raf-1), ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2). Results The survival rates of U251 cells at GLE concentrations of 0, 50, 100, 200 mg/L were 100%, (86.62±4.26)%, (67.68±3.49)%, (50.84±3.39)%, and (40.13±3.25)%, re-spectively, with statistically significant differences (P<0.05). The difference of the apoptosis rate, colony formation number, scratchhealing rate, invasive cell number, PBX3, ERK mRNA and the relative protein expression levels of PBX3, p-Raf-1 and p-ERK1/2 of U251 cells were statistically significant at 0, 50, 100, 200 mg/L GLE (P<0.05). The survival rate, scratch healing rate, PBX3 and ERK mRNA relative expression levels and levels of RAS, PBX3, p-Raf-1, p-MEK1/2 and p-ERK1/2 protein relative expression decreasedwith the increase of GLE concentration. The number of colony formation and invasive cells decreased, and the apoptosis rate increasedwith the increase of GLE concentration. The effect of GLE was dose-dependent (P<0.05). Conclusion GLE can inhibit the prolifera-tion, clonal formation, migration, invasion and other malignant biological behaviors of glioma cells, and promote apoptosis, which maybe related to the inhibition of PBX3/MAPK axis activation. |
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