文章摘要
王国俊,王灼印,朱天宇,高不郎,李瑞欣,王敬涛,张云飞,丁恒轩.华蟾素通过下调PIM3的表达抑制高转移胃癌细胞的实验研究[J].,2021,(3):436-442
华蟾素通过下调PIM3的表达抑制高转移胃癌细胞的实验研究
Experimental Study of Bufothionine Inhibits Highly Metastatic Gastric Cancer Cells by Down Regulating PIM3 Expression
投稿时间:2020-08-23  修订日期:2020-09-18
DOI:10.13241/j.cnki.pmb.2021.03.007
中文关键词: 胃癌  华蟾素  丁硫磷  PIM3  细胞活性
英文关键词: Gastric cancer  Cinobufacini  Bufothionine  PIM3  Cell activity
基金项目:河南省高等学校重点科研计划项目(17A320045)
作者单位E-mail
王国俊 郑州大学第一附属医院胃肠外科 河南 郑州 450022 fccwanggj@zzu.edu.cn 
王灼印 郑州大学第一附属医院胃肠外科 河南 郑州 450022  
朱天宇 郑州大学第一附属医院胃肠外科 河南 郑州 450022  
高不郎 郑州大学第一附属医院胃肠外科 河南 郑州 450022  
李瑞欣 郑州大学第一附属医院胃肠外科 河南 郑州 450022  
王敬涛 郑州大学第一附属医院胃肠外科 河南 郑州 450022  
张云飞 郑州大学第一附属医院胃肠外科 河南 郑州 450022  
丁恒轩 郑州大学第一附属医院胃肠外科 河南 郑州 450022  
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中文摘要:
      摘要 目的:癌(GC)是全球第四大常见癌症。丁硫磷是华蟾酥的主要活性成分,从蟾蜍的皮肤和腮腺毒腺中提取,在体外具有抗癌活性。然而,丁硫磷是否对胃癌有抗癌作用尚不清楚。本研究旨在评价丁硫磷对GC的抑制作用。方法:以高转移性MKN28 GC细胞为细胞模型,研究华蟾素的抗癌作用。使用CCK-8测定细胞活力,LDH检测丁硫磷对细胞膜完整性的影响。Hoechst/PI双染色和Annexin V-FITC/PI染色检测细胞凋亡,随后使用流式细胞仪进行分析。western blotting检测相关蛋白的表达水平。通过iTRAQ分析,确定丁硫磷处理后GC细胞的差异表达基因。结果:丁硫磷能抑制高转移性GC MKN28细胞的生长,破坏细胞膜,促进GC细胞凋亡。iTRAQ分析表明PIM3是丁硫磷素抗癌作用的分子靶点。研究还发现,PIM3基因敲除显著增强了丁硫磷对GC细胞的抗生长和促凋亡作用。相反,异位PIM3的表达明显抑制了丁硫磷的抗肿瘤活性。结论:华蟾素对高转移性的GC具有抗癌活性是通过下调PIM3表达而实现的。
英文摘要:
      ABSTRACT Objective: Gastric cancer (GC) is the fourth most common cancer globally. Bufothionine is a major active constituent of Cinobufacini, which is extracted from the skin and parotid venom gland of the toad Bufo bufogargarizans Cantor. It exhibits anti-cancer activities in vitro. However, whether bufothionine exerts anti-cancer activities against GC is unknown. To evaluate the inhibitory effect of bufothionine on GC. Methods: MKN28 GC cells were chosen as cell models to study the anti-cancer effect of bufothionine. Cell viability was determined by CCK-8 assay, while the effect of bufothionine on cell membrane integrity was examined by LDH assay. Cell apoptosis was detected by Hoechst/PI staining and Annexin V-FITC/PI staining followed by flow cytometry analysis. The expression levels of proteins involved were examined using western blotting. iTRAQ analysis was conducted to identify the differentially expressed genes in AGS cells following bufothionine treatment. Results: The results revealed that bufothionine prevented the growth, destroyed cell membrane, and promoted apoptotic cell death of GC cells. iTRAQ analysis revealed that PIM3 might be a molecular target responsible for the anti-cancer effects of bufothionine. PIM3 knockdown significantly augmented the anti-growth and pro-apoptotic effects of bufothionine in GC cells. In contrast, ectopic PIM3 expression markedly dampened the anti-neoplastic activities of bufothionine. Conclusion: Bufothionine exhibits anti-cancer activities in vitro GC cells via downregulating PIM3.
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