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Activated mitochondrial apoptosis in hESCs after dissociation involving the PKA/p-p53/Bax signaling pathway
Zhang, Liang1,3; Ma, Liang1,2; Yan, Tingxuan1,2; Han, Xinya2; Xu, Jiandong3; Xu, Jian1,2; Xu, Xia1,2,3
2018-08-15
Source PublicationEXPERIMENTAL CELL RESEARCH
ISSN0014-4827
Volume369Issue:2Pages:226-233
Abstract

Human embryonic stem cells (hESCs) are highly fragile with massive cell death after dissociation into single cells, which seriously hampers their applications. The mechanism underlying the massive cell death after dissociation still remains elusive. Here, the expression of apoptosis-related proteins, cell survival and mitochondrial membrane potential in dissociated hESCs before and after the treatments with a protein kinase A (PKA) inhibitor H89 and p53 inhibitor Pifithrin alpha were investigated, respectively. Protein interactions were identified by immunoprecipitation and immunofluorescence. The results show that the dissociation causes Caspase-dependent apoptosis in hESCs mediated by mitochondrial pathway with the up-regulation of pro-apoptotic proteins, decrease in mitochondrial membrane potential and elevation in pro-apoptotic Cyto c release, which are obviously suppresses by H89. The dissociation-induced increase of phosphorylated p53 Ser15 (p-p53) is suppressed by Pifithrin a which also rescues the elevated levels of pro-apoptotic proteins in mitochondrial pathway. During the dissociation-induced apoptosis, PKA/p-p53/Bax signaling pathway is identified by immunoprecipitation and immunofluorescence showing the most likely interaction between them. These results indicate that dissociation induces mitochondrial apoptosis in hESCs involving PKA/p-p53/Bax signaling pathway, which not only give new insights into the apoptosis mechanism of dissociated hESCs, but also provide clues for developing potential strategies to promote hESC survival after dissociation.

KeywordHuman Embryonic Stem Cells Dissociation Mitochondrial Apoptosis Pathway Pka P-p53
DOI10.1016/j.yexcr.2018.05.024
Language英语
WOS KeywordEmbryonic Stem-cells ; Outer-membrane Permeabilization ; Protein-kinase-a ; Dependent Apoptosis ; Trail Receptors ; Wild-type ; P53 ; Survival ; Death ; Inhibitor
Funding ProjectNational Natural Science Foundation of China[21576266] ; National Natural Science Foundation of China[21606239] ; Major Research Plan of the National Natural Science Foundation of China[91534107] ; Wanjiang Scholar Program from Anhui University of Technology ; Start Fund for Biochemical Engineering Research Center from Anhui University of Technology
WOS Research AreaOncology ; Cell Biology
WOS SubjectOncology ; Cell Biology
Funding OrganizationNational Natural Science Foundation of China ; Major Research Plan of the National Natural Science Foundation of China ; Wanjiang Scholar Program from Anhui University of Technology ; Start Fund for Biochemical Engineering Research Center from Anhui University of Technology
WOS IDWOS:000442063200006
PublisherELSEVIER INC
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/25531
Collection中国科学院过程工程研究所
Corresponding AuthorXu, Xia
Affiliation1.Anhui Univ Technol, Biochem Engn Res Ctr, 59 Hudong Rd, Maanshan 243002, Anhui, Peoples R China
2.Anhui Univ Technol, Sch Chem & Chem Engn, Maanshan 243002, Anhui, Peoples R China
3.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100190, Peoples R China
Recommended Citation
GB/T 7714
Zhang, Liang,Ma, Liang,Yan, Tingxuan,et al. Activated mitochondrial apoptosis in hESCs after dissociation involving the PKA/p-p53/Bax signaling pathway[J]. EXPERIMENTAL CELL RESEARCH,2018,369(2):226-233.
APA Zhang, Liang.,Ma, Liang.,Yan, Tingxuan.,Han, Xinya.,Xu, Jiandong.,...&Xu, Xia.(2018).Activated mitochondrial apoptosis in hESCs after dissociation involving the PKA/p-p53/Bax signaling pathway.EXPERIMENTAL CELL RESEARCH,369(2),226-233.
MLA Zhang, Liang,et al."Activated mitochondrial apoptosis in hESCs after dissociation involving the PKA/p-p53/Bax signaling pathway".EXPERIMENTAL CELL RESEARCH 369.2(2018):226-233.
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