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PTS通过调控Wnt/β-catenin通路对胃癌细胞的影响
Effects of PTS on gastric cancer cells by regulating Wnt/β-catenin pathway
收稿日期:  
DOI:10.3969/j.issn.1673-9701.2024.19.006
关键词:  6-丙酮酰四氢蝶呤合成酶  胃癌  Wnt/β-catenin信号通路  恶性生物学行为
Key Words:
基金项目:山东省潍坊市2021年科学技术发展计划项目(2021YX105);潍坊医学院研究生科研创新基金项目(2022-28)
作者单位
杨金鹏 山东第二医科大学临床医学院山东潍坊 261042 
王文华 山东第二医科大学临床医学院山东潍坊 261042 
张永博 山东第二医科大学临床医学院山东潍坊 261042 
张玉英 潍坊市人民医院消化内科山东潍坊 261000 
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摘要:目的 探讨6-丙酮酰四氢蝶呤合成酶(6-pyruvoyl-tetrahydropterin synthase,PTS)对胃癌细胞的影响。方法 通过蛋白质印迹法(Western blot,WB)和逆转录定量聚合酶链反应(reverse transcription quantitative polymerase chain reaction,RT-qPCR)测定胃黏膜上皮细胞GES-1及胃癌细胞系HGC-27细胞、MGC-803细胞、AGS细胞、MKN-45细胞中PTS蛋白及mRNA的表达。选取AGS细胞进行后续实验,将AGS细胞分为对照(NC)组、sh-PTS组、sh-PTS+BML-284组。采用CCK-8测定细胞增殖;采用划痕愈合测定细胞迁移能力;采用Transwell实验测定细胞迁移、侵袭能力;采用流式细胞术检测细胞凋亡;WB和RT-qPCR检测细胞中β-catenin、c-myc、GSK-3β、Wnt5a蛋白和mRNA水平。结果 胃黏膜上皮细胞GES-1中PTS表达低于胃癌细胞,AGS细胞中PTS表达最高;敲低PTS可抑制胃癌细胞增殖、迁移、侵袭,促进胃癌细胞凋亡,β-catenin、c-myc、Wnt5a蛋白水平下降,GSK-3β蛋白水平升高(P<0.05);与sh-PTS组相比,sh-PTS+BML-284组细胞增殖、迁移、侵袭能力增强,细胞凋亡水平下降,β-catenin、c-myc、Wnt5a蛋白表达升高,GSK-3β蛋白表达下降(P<0.05)。结论 PTS通过调控Wnt/β-catenin通路影响胃癌细胞的增殖、迁移、侵袭及凋亡。
Abstract:Objective To investigate the effects of 6-pyruvoyl-tetrahydropterin synthase (PTS) on gastric cancer cells. Methods The expression of PTS protein and mRNA in gastric mucosal epithelial cells GES-1 and gastric cancer cell lines HGC-27 cells, MGC-803 cells, AGS cells and MKN-45 cells were determined by Western blot (WB) and reverse transcription quantitative polymerase chain reaction (RT-qPCR). AGS cells were selected for follow-up experiments. AGS cells were divided into control (NC) group, sh-PTS group, and sh-PTS+BML-284 group. Cell proliferation was measured by CCK-8. Cell migration ability was measured by scratch healing. Cell invasion and migration were measured by Transwell assay. Apoptosis was measured by flow cytometry. The protein and mRNA levels of β-catenin, c-myc, GSK-3β and Wnt5a were detected by WB and RT-qPCR. Results The expression of PTS in gastric mucosal epithelial cells GES-1 was lower than that in gastric cancer cells, and the expression of PTS was the highest in AGS cells. Knocking down PTS could inhibit proliferation, migration and invasion of gastric cancer cells, promote cell apoptosis, decreased β-catenin, c-myc, Wnt5a protein levels, and increased GSK-3β protein levels (P<0.05). Compared with sh-PTS group, cell proliferation, migration and invasion ability of sh-PTS+BML-284 group were enhanced, apoptosis level was decreased, protein expression of β-catenin, c-myc and Wnt5a was increased, and protein expression of GSK-3β was decreased (P<0.05). Conclusion PTS can affect proliferation, migration, invasion and apoptosis of gastric cancer cells by mediating Wnt/β-catenin pathway.
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