摘要
目的探讨南蛇藤茎醋酸乙酯提取物(COE)通过提高丝氨酸羟甲基转移酶2(SHMT2)琥珀酰化水平影响胃癌细胞增殖、凋亡和丝氨酸/甘氨酸代谢的作用。方法采用MTT法、TUNEL试剂盒检测COE对MGC-803细胞增殖和凋亡的影响。通过免疫沉淀富集MGC-803细胞中SHMT2蛋白,利用琥珀酰化泛抗体检测SHMT2琥珀酰化修饰水平。将细胞裂解液进行~1H-NMR检测,结合多元统计分析比较阴性对照组和320μg/ml COE组丝氨酸/甘氨酸含量差异。结果 MTT法结果显示,20、40、80、160、320μg/ml COE分别处理MGC-803细胞24、48、72 h后,细胞的增殖活性受到明显抑制,并呈浓度和时间依赖性。经COE作用后,细胞核固缩,胞浆浓缩,胞膜破裂,出现凋亡小体,随着COE作用浓度升高,细胞凋亡明显。320μg/ml COE组细胞琥珀酰化SHMT2蛋白表达量高于阴性对照组(0. 92±0. 10 vs. 0. 18±0. 05,P<0. 05)。阴性对照组和320μg/ml COE组细胞丝氨酸(7. 861±0. 367 vs. 2. 594±0. 037)和甘氨酸(12. 915±0. 581 vs. 4. 620±0. 382)含量差异具有统计学意义(P<0. 05)。结论 COE可通过影响胃癌细胞MGC-803丝氨酸/甘氨酸代谢,抑制其增殖,并诱导其凋亡,其可能的作用机制与上调丝氨酸代谢关键酶SHMT2的琥珀酰化水平有关。
Objective To discuss the effect and mechanism of celastrus orbiculatus extract(COE)on serine/glycine metabolism,proliferation and apoptosis of gastric cancer via up-regulating the succinylation of serine hydroxyl methyl transferase 2(SHMT2).Methods The effect of COE on proliferation and apoptosis of MGC-803 cells was detected by MTT assay and TUNEL.SHMT2 protein in MGC-803 cells was enriched by immunoprecipitation,and the level of succinylated modification of SHMT2 was detected by succinylated pan-antibody.The cell lysis fluid was detected by 1H-NMR,and the difference in serine/glycine content between the negative control group and the 320μg/ml COE group was compared by multivariate statistical analysis.Results The results of MTT showed that the proliferation of MGC-803 cells treated with 20,40,80,160 and 320μg/ml COE for 24,48 and 72 hours respectively was inhibited obviously,and showed a concentration and time dependence.After COE treatment,the nucleus was condensed,the cytoplasm was concentrated,the cell membrane was ruptured,and apoptotic bodies appeared.With the increase of COE treatment concentration,cell apoptosis was obvious.The expression level of succinylated SHMT2 protein in 320μg/ml COE group was higher than that in the negative control group(0.92±0.10 vs.0.18±0.05,P<0.05).The difference of serine(7.861±0.367 vs.2.594±0.037)and glycine(12.915±0.581 vs.4.620±0.382)between the negative control group and 320μg/ml COE group was statistically significant(P<0.05).Conclusion COE can inhibit proliferation and induce apoptosis of gastric cancer cells by affecting the metabolism of MGC-803 serine/glycine,and its possible mechanism of action is related to the up-regulation of the succinylation level of SHMT2,a key enzyme of serine metabolism.
作者
彭微
陈琳
刘洁
张爱武
PENG Wei;CHEN Lin;LIU Jie;ZHANG Aiwu(Department of Basic Medicine,Hunan Traditional Chinese Medical College,Zhuzhou 412000,China)
出处
《临床肿瘤学杂志》
CAS
北大核心
2020年第6期481-486,共6页
Chinese Clinical Oncology
基金
湖南省自然科学基金资助项目(2019JJ60072)
湖南省中医药科研计划资助项目(201927)。
关键词
胃癌
南蛇藤茎醋酸乙酯提取物
丝氨酸羟甲基转移酶2
丝氨酸/甘氨酸代谢
增殖
凋亡
Gastric cancer
Celastrus orbiculatus extract(COE)
Serine hydroxyl methyl transferase 2(SHMT2)
Ser/Gly metabolites
Proliferation
Apoptosis
