蛋白质琥珀酰化参与丁酸抑制胃癌细胞增殖及迁移的作用  

Protein succinylation involved in the suppression of gastric cancer cell proliferation and migration by butyric acid

作　　者：黄颖鹏[1] 张珂 吴芳全 施迪邦 韩千年 王方岩 HUANG Yingpeng;ZHANG Ke;WU Fangquan;SHI Dibang;HAN Qiannian;WANG Fangyan(Department of Gastrointestinal Surgery,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China;School of Basic Medicine Science,Wenzhou Medical University,Wenzhou 325035,China;Department of Gastroenterology,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院胃肠外科,浙江温州325027 [2]温州医科大学基础医学院,浙江温州325035 [3]温州医科大学附属第二医院育英儿童医院消化内科,浙江温州325027

出　　处：《温州医科大学学报》2024年第2期99-105,共7页Journal of Wenzhou Medical University

基　　金：浙江省公益技术研究计划项目(LGF20H070003);温州市基础性科研项目(Y2020214,Y20190060)。

摘　　要：目的:研究丁酸对胃癌细胞增殖及代谢的影响,并探讨其潜在作用机制。方法:使用5 mmol/L丁酸钠处理胃癌细胞48 h后,通过EdU染色及Transwell法检测细胞增殖、迁移情况;收集丁酸处理后的胃癌细胞,提取代谢物进行代谢组学分析筛选出其中显著富集的代谢通路以及上调的代谢物;采用Western blot以及细胞免疫荧光检测丁酸钠处理后胃癌细胞蛋白质琥珀酰化程度,同时采用RT-qPCR检测SIRT5、SUCLG1、SUCLG2和SUCLA2的mRNA水平。结果:丁酸钠处理48 h后胃癌细胞增殖被显著抑制,细胞中检测到三羧酸循环及氧化磷酸化等代谢通路显著富集,其中三羧酸循环代谢物L-苹果酸、琥珀酰辅酶A及延胡索酸显著上调(均P<0.05);Western blot和免疫荧光显示丁酸处理后胃癌细胞蛋白琥珀酰化水平上调(P<0.05)。同时RT-qPCR证实了丁酸处理上调了琥珀酰辅酶A合成酶编码基因SUCLG1、SUCLG2和SUCLA2的mRNA表达水平(均P<0.05),但并未影响SIRT5表达(P>0.05)。结论:丁酸能够抑制胃癌细胞增殖及迁移,其可能通过促进SUCLs表达以及上调琥珀酰辅酶A水平并进一步促进胃癌细胞蛋白质琥珀酰化实现。Objective:To study the effects of butyrate on the proliferation and metabolism of gastric cancer cells,and explore its potential mechanism.Methods:EdU and Transwell were used to detect the proliferation and migration of gastric cancer cells after 5 mmol/L sodium butyrate treatment for 48 h,and the cells were collected and metabolites were extracted for metabolomic analysis.Then we screened out the significantly enriched metabolic pathways and upregulated metabolites.Western blot and cellular immunofluorescence were used to detect the degree of protein succinylation in gastric cancer cells treated with sodium butyrate.Meanwhile,the mRNA level of SIRT5,SUCLG1,SUCLG2,and SUCLA2 was detected by RT-qPCR.Results:After 48 h of sodium butyrate treatment,the proliferation of gastric cancer cells decreased significantly.The metabolic pathways such as“TCA cycle”and“oxidative phosphorylation”were significantly enriched in the treatment group,and the TCA cycle metabolite L-Malic acid,succinyl-CoA and fumarate were significantly upregulated(all P<0.05).Western blot and immunofluorescence showed that the protein succinylation of gastric cancer cells was activated after sodium butyrate treatment(P<0.05).What’s more,RT-qPCR results confirmed that sodium butyrate treatment increased the mRNA expression of succinyl-CoA synthetase encoding gene,including SUCLG1,SUCLG2,and SUCLA2(all P<0.05),but did not significantly affect the expression of SIRT5(P<0.05).Conclusion:Butyrate inhibits the proliferation of gastric cancer cells by activating SUCLs expression and increasing the level of succinyl-CoA to further promote the protein succinylation of gastric cancer cells.

关 键 词：丁酸 胃肿瘤 代谢组学 琥珀酰化 琥珀酰辅酶A 

分 类 号：R735.2[医药卫生—肿瘤]