Bi Yakun, Wu Daoyan, Wu Xiaojuan, Wang Fei, Yu Hang, Liu Pan, Cui Guzhen, Chen Zhenghong
Key Laboratory of Microbiology and Parasitology of Education Department of Guizhou, School of Basic Medical Science, Guizhou Medical University, Guiyang, China.
Department of Clinical Laboratory, The Maternal and Child Health Care Hospital of Guizhou Medical University, Guizhou Medical University, Guiyang, China.
Ann Transl Med. 2022 Feb;10(4):176. doi: 10.21037/atm-21-7045.
Reactive oxygen species (ROS)-induced oxidative stress (OS) and hyper-proliferation of gastric epithelial cells (GECs) due to (Hp) infection are important mechanisms that lead to gastric carcinoma. Phycocyanin is a marine functional food additive with antioxidant and anti-inflammatory properties.
The flow cytometry was used to detect the effect of 150 µM phycocyanin intervention on the cell cycle of human gastric adenocarcinoma cell line (AGS) infected with Hp. The transcriptomics of AGS cells intervened by 150 µM phycocyanin for 24 h and infected by Hp were detected. Differential gene expression analysis was performed using a cutoff at the normalized gene expression (log2) of 2 and a P-value of <0.05. Comparisons of the transcriptomes were made between the following groups: (I) AGS cells not infected with Hp and not using phycocyanin action and AGS cells infected with Hp only; (II) AGS cells not infected with Hp and not using phycocyanin action and AGS cells infected with phycocyanin action only; and (III) AGS cells infected with Hp only and phycocyanin treated and infected with Hp cells. c-myc and CyclinD1 was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and immunoblotting. Phosphorylation and non-phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 Mitogen-activated protein kinase (p38MAPK) were detected by immunoblotting. Intracellular ROS was detected by immunofluorescence.
Phycocyanin alleviates the Hp infection-induced increased cell viability and expression of cell cycle regulatory proteins c-myc and CyclinD1. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that the differentially expressed genes in phycocyanin-treated Hp-infected AGS cells were most significantly enriched in the mitogen-activated protein kinase (MAPK) signaling pathway. Phycocyanin could inhibit the Hp infection-induced phosphorylation of JNK, ERK, and p38MAPK and reduce the level of cellular ROS.
This study suggests that phycocyanin can regulate the ROS/MAPK signaling pathway and reduce c-myc and CyclinD1 expression to inhibit the hyper-proliferation of AGS cells. Phycocyanin may serve as an inhibitor of malignant progression of Hp infection-induced gastric disease.
活性氧（ROS）诱导的氧化应激（OS）以及幽门螺杆菌（Hp）感染导致的胃上皮细胞（GECs）过度增殖是引发胃癌的重要机制。藻蓝蛋白是一种具有抗氧化和抗炎特性的海洋功能性食品添加剂。
采用流式细胞术检测150 μM藻蓝蛋白干预对感染Hp的人胃腺癌细胞系（AGS）细胞周期的影响。检测150 μM藻蓝蛋白干预24 h并感染Hp的AGS细胞的转录组学。使用标准化基因表达（log2）截断值为2且P值<0.05进行差异基因表达分析。在以下几组之间进行转录组比较：（I）未感染Hp且未使用藻蓝蛋白作用的AGS细胞与仅感染Hp的AGS细胞；（II）未感染Hp且未使用藻蓝蛋白作用的AGS细胞与仅使用藻蓝蛋白作用的AGS细胞；以及（III）仅感染Hp的AGS细胞与经藻蓝蛋白处理并感染Hp的细胞。通过定量实时聚合酶链反应（qRT-PCR）和免疫印迹检测c-myc和细胞周期蛋白D1（CyclinD1）。通过免疫印迹检测c-Jun氨基末端激酶（JNK）、细胞外信号调节激酶（ERK）和p38丝裂原活化蛋白激酶（p38MAPK）的磷酸化和非磷酸化状态。通过免疫荧光检测细胞内ROS。
藻蓝蛋白减轻了Hp感染诱导的细胞活力增加以及细胞周期调节蛋白c-myc和CyclinD1的表达。京都基因与基因组百科全书（KEGG）富集分析表明，经藻蓝蛋白处理的感染Hp的AGS细胞中差异表达基因最显著富集于丝裂原活化蛋白激酶（MAPK）信号通路。藻蓝蛋白可抑制Hp感染诱导的JNK、ERK和p38MAPK的磷酸化，并降低细胞内ROS水平。
本研究表明藻蓝蛋白可调节ROS/MAPK信号通路，降低c-myc和CyclinD1表达，从而抑制AGS细胞的过度增殖。藻蓝蛋白可能作为Hp感染诱导的胃部疾病恶性进展的抑制剂。
