首页|MiR-155靶向talin-1基因调控柱状黄杆菌胞外多糖诱导的EPC细胞凋亡

MiR-155靶向talin-1基因调控柱状黄杆菌胞外多糖诱导的EPC细胞凋亡

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为研究miR-155在柱状黄杆菌胞外多糖(FC-EPS)诱导的细胞凋亡中的作用机制，本实验采用RNA干扰(RNAi)技术，通过过表达miR-155和敲降踝蛋白基因(talin-1)，皆能抑制FC-EPS诱导的细胞凋亡，并鉴定出talin-1为miR-155的靶蛋白。在FC-EPS诱导细胞凋亡的过程中，踝蛋白水平显著升高，在凋亡发生阶段检测到凋亡执行蛋白Caspase-3的活化，同时检测到2条踝蛋白剪切异构体，大小分别约为200 ku和250ku;将构建的鲤Talin-1真核表达质粒转染鲤上皮瘤细胞(EPC)，过表达的Talin-1延迟且延长了细胞凋亡的进程，并检测到上述2条踝蛋白异构体。然而，当以FC-EPS转染EPC细胞时，Talin-1先下降再恢复到正常水平，细胞不发生凋亡。因此，在FC-EPS诱导细胞凋亡的过程中，细胞通过上调miR-155来调控靶基因talin-1 mRNA和蛋白转录的变化，以及产生Talin-1蛋白异构体来抑制细胞发生凋亡。本研究为有效防治柱状黄杆菌疾病提供新思路。
MiR-155 regulates apoptosis induced by FC-EPS incubation via targeting the talin-1 gene in EPC
Incubation of Flavobacterium columnare exopolysaccharide(FC-EPS)significantly increases the expression of miR-155 in epithelioma papulosum cyprinid(EPC)cell line,leading to cell apoptosis.In order to study the mechanism of miR-155 in FC-EPS induced apoptosis,RNAi technology was used in this research.It was found that both overexpression of miR-155 and talin-1 knockout could inhibit apoptosis induced by FC-EPS incub-ation.Then,talin-1 was identified as the target protein of miR-155 by EMSA assay.During the process of apop-tosis induced by FC-EPS incubation,the Talin-1 protein was significantly increased,and the activation of apop-tosis executioner caspase-3 protein was detected.At the same time,two talin-1 isomers(about 200 ku and 250 ku,respectively)were obtained.Subsequently,the talin-1 gene inserted in plasmid pcDNA3.1 was constructed and transfected into EPC cells,the overexpressed Talin-1 protein delayed and prolonged the process of apoptosis,and two Talin-1 isomers mentioned above were detected.However,apoptosis did not occur when EPC cells were trans-fected with FC-EPS;on this occasion,the Talin-1 protein in EPC went down at first,then returned to normal level.Hence,in the context of apoptosis induced by FC-EPS incubation,EPC cells reduces apoptosis by regulating the level of mRNA and protein transcription of talin-1 through up-regulation of miR-155 and producing Talin-1 iso-mers.This paper provides a new idea for effective prevention and treatment of F.columnare diseases.

Flavobacterium columnaremicroRNA-155talin-1apoptosisRNA interference

王骞、王承德、罗敏意、黎果、车行、林蠡、赵丽娟

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仲恺农业工程学院动物科技学院,广东广州 510225

中山市小榄镇农业服务中心,广东中山 510230

高州市百联水产种苗有限公司,广东茂名 525200

柱状黄杆菌 miR-155 talin-1 细胞凋亡 RNA干扰

国家自然科学基金中山市社会公益与基础研究项目(第二批)(2022)广东省科技创新和乡村振兴战略专项(2021)广东省现代农业产业技术体系建设项目创新团队水产疫病监测与综合防控共性关键技术研发创新团队项目

316021902022B20032021S00822019KJ141

2024

10.11964/jfc.20240214389

水产学报
中国水产学会

水产学报

CSTPCD北大核心
影响因子：1.148
ISSN：1000-0615
年,卷(期)：2024.48(5)
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