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华中师范大学学报(自然科学版)  2024, Vol. 58 Issue (2): 214-223    
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伤寒沙门菌外膜囊泡通过调控铁死亡抑制结直肠癌细胞的增殖及机制初探
戈艺潼1, 吉 滢1,2, 曾敏敏1, 郑学明1,2, 黄新祥1, 张 盈1
(1.江苏大学医学院, 江苏 镇江 212013; 2.江苏大学消化疾病研究所, 江苏 镇江 202000)
Inhibitory effectand mechanism of Salmonella Typhi-outer membrane vesicles on the proliferation of colorectal cancer cells by regulating ferroptosis
GE Yitong1, JI Ying1,2, ZENG Minmin1, ZHENG Xueming1,2, HUANG Xinxiang1,ZHANG Ying1
(1.School of Medicine, Jiangsu University, Zhenjiang 212013, Jiangsu, China;2.Institute of Digestive Diseases, Jiangsu University, Zhenjiang 212013, Jiangsu, China)
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摘要 近年来,细菌外膜囊泡(OMVs)在抗肿瘤治疗中展现出的巨大潜力引起了广泛的关注.为探究伤寒沙门菌外膜囊泡(S. Typhi-OMVs)对人结直肠癌细胞株HT-29增殖的影响及可能的作用机制,通过超速离心法提取不同细菌的OMVs,细胞增殖实验(CCK-8)检测各细菌OMVs对细胞活力的影响;转录组测序(RNA-seq)分析处理后细胞基因表达水平的变化;试剂盒检测铁死亡相关标志物的含量变化;实时荧光定量PCR(qRT-PCR)和蛋白质免疫印迹法(western blot)分别检测相关基因mRNA和蛋白质的表达变化.结果显示,在提取的6种细菌OMVs中,S. Typhi-OMVs对HT-29细胞的增殖产生了最明显的抑制作用,并且呈现出浓度和时间梯度依赖性;RNA-seq显示HT-29细胞可能发生了铁死亡;S. Typhi-OMVs作用后,细胞内发生了铁沉积,氧化产物增多,抗氧化剂减少,符合铁死亡生化特征;SAT1是S. Typhi-OMVs处理后HT-29胞内mRNA表达量变化最大的基因;p53-SAT1-ALOX15是铁死亡的信号通路之一,S. Typhi-OMVs处理后胞内p53、SAT1、ALOX15的mRNA与蛋白表达均增高.以上结果表明,S. Typhi-OMVs能够抑制结直肠癌细胞HT-29的增殖,其机制可能与通过p53-SAT1-ALOX15信号通路诱导HT-29发生铁死亡有关.
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戈艺潼
吉 滢
曾敏敏
郑学明
黄新祥
张 盈
关键词 铁死亡外膜囊泡伤寒沙门菌结直肠癌细胞增殖    
Abstract:In recent years, the tremendous potential of bacterial outer membrane vesicles (OMVs) in anti-tumor therapy has attracted wide attention. To investigate the effect of Salmonella enterica serovar Typhi outer membrane vesicles (S. Typhi-OMVs) on the proliferation of human colorectal cancer cell line HT-29 and the possible mechanism of action, the outer membrane vesicles of different bacteria were extracted by ultracentrifugation. Cell proliferation assay (CCK-8) was performed to detect the effect of each bacterial outer membrane vesicles on HT-29 cell viability. Transcriptome sequencing was used to analyze the changes of gene expression levels in HT-29 cells after S. Typhi-OMVs treatment. The content changes of ferroptosis related markers were detected with the corresponding kits. Quantitative real-time polymerase chain reaction and western blot were applied to detect the changes of mRNA and protein expression of related genes, respectively. The results showed that S. Typhi-OMVs had the most significant inhibitory effect on the proliferation of HT-29 among the six bacterial outer membrane vesicles, and the inhibitory effect showed concentration and time gradient dependence. RNA-seq revealed that ferroptosis may have occurred in HT-29. After S. Typhi-OMVs treatment, intracellular iron deposition occurred, oxidative products increased, and antioxidants decreased, which was consistent with the biochemical characteristics of ferroptosis. SAT1 was the gene with the largest mRNA expression change. The p53-SAT1-ALOX15 is one of the signaling pathways of ferroptosis, and the mRNA and protein expressions of p53, SAT1 and ALOX15 were all increased after S. Typhi-OMVs treatment of HT-29. The above results suggest that S. Typhi-OMVs were able to inhibit the proliferation of colorectal cancer cells HT-29, and the mechanism might be related to the induction of ferroptosis in HT-29 through the p53-SAT1-ALOX15 signaling pathway.
Key wordsferroptosis    outer membrane vesicles    Salmonella enterica serovar Typhi    colorectal cancer    cell proliferation
收稿日期: 2024-04-07     
引用本文:   
戈艺潼,吉 滢,曾敏敏,郑学明,黄新祥,张 盈. 伤寒沙门菌外膜囊泡通过调控铁死亡抑制结直肠癌细胞的增殖及机制初探[J]. 华中师范大学学报(自然科学版), 2024, 58(2): 214-223.
GE Yitong,JI Ying,ZENG Minmin,ZHENG Xueming,HUANG Xinxiang,ZHANG Ying. Inhibitory effectand mechanism of Salmonella Typhi-outer membrane vesicles on the proliferation of colorectal cancer cells by regulating ferroptosis. journal1, 2024, 58(2): 214-223.
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https://journal.ccnu.edu.cn/zk/CN/     或     https://journal.ccnu.edu.cn/zk/CN/Y2024/V58/I2/214
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