Detection of Apoptosis Based on the Interaction between Annexin V and Phosphatidylserine

Liu, Ting; Zhu, Wei; Yang, Xiang; Chen, Lin; Yang, Rongwu; Zhang, Hua; Li, Genxi

doi:10.1021/ac801267s

Cited by 70 publications

(50 citation statements)

References 21 publications

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“…In the present study it was found that treatment with TGF-β1 (10 and 20 ng/ml) significantly upregulated Bax but downregulated Bcl-2 protein expression in BMSCs. Annexin V is another apoptotic promoter that preferentially binds phosphatidylserine and promotes cell apoptosis (30). The present results demonstrated that Annexin V was markedly upregulated in BMSCs following exposure to 10 and 20 ng/ml TGF-β1 for 24 h.…”

Section: Discussionsupporting

confidence: 55%

TGF-β1 induces apoptosis of bone marrow-derived mesenchymal stem cells via regulation of mitochondrial reactive oxygen species production

Zhang

Ren

2015

Experimental and Therapeutic Medicine

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Abstract. Bone marrow-derived mesenchymal stem cells (BMSCs) are the most promising seed cells in regenerative medicine. Our previous study demonstrated that transforming growth factor (TGF)-β1 induced BMSC senescence in vitro. Whether TGF-β1 affects the apoptosis of BMSCs has not been examined; therefore the aim of the present study was to investigate this effect. BMSCs were isolated from mouse bone marrow, and the third-passage cells were exposed to 0, 10 and 20 ng/ml TGF-β1 for 24 h. Cell proliferation was measured by MTT assay; apoptosis was assessed using DAPI staining; and the apoptotic signals Annexin V, B-cell lymphoma (Bcl)-2 and Bcl-2-associated X protein (Bax) were measured using western blotting. Mitochondrial reactive oxygen species (ROS) were measured by flow cytometry following staining with MitoSOX™ Red mitochondrial superoxide indicator. The MTT assay showed that 10 and 20 ng/ml TGF-β1 inhibited BMSC proliferation. DAPI staining demonstrated that 10 and 20 ng/ml TGF-β1 promoted BMSC apoptosis, which was further confirmed by a western blotting assay showing a significant increase in the pro-apoptotic signals Annexin V and Bax but a decrease in the anti-apoptotic signal Bcl-2. It was also found that TGF-β1 markedly increased the mitochondrial ROS levels in BMSCs. It is well known that mitochondrial ROS are strong stimulators of cell apoptosis. These findings indicate that TGF-β1 can induce BMSC apoptosis, and the mechanism may involve mitochondrial ROS generation.

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Section: Discussionsupporting

confidence: 55%

TGF-β1 induces apoptosis of bone marrow-derived mesenchymal stem cells via regulation of mitochondrial reactive oxygen species production

Zhang

Ren

2015

Experimental and Therapeutic Medicine

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“…Estimates of apoptosis rates of H9c2 cells widely make use of FITC-AV/PI double staining analyzed by flow cytometry (Shang et al, 2013). The annexin V assay is sensitive and supplies the possibility of detecting early phases of apoptosis before the loss of cell membrane integrity, and annexin V in polyethylenimine film can hold its high affinity following with phosphatidylserine altered from the inner to the outer cell membrane of the apoptotic cells in in vitro experiments (Vermes et al, 1995;Liu et al, 2009). In this study, the cardiomyocyte protection of HA+GA may be associated with the suppression of the cell apoptosis through the decreased apoptotic rates.…”

Section: Discussionmentioning

confidence: 76%

Protective mechanisms of hypaconitine and glycyrrhetinic acid compatibility in oxygen and glucose deprivation injury

Wang

Wan

Zhou

et al. 2017

J. Zhejiang Univ. Sci. B

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This study investigated the protective effect of the compatibility of hypaconitine (HA) and glycyrrhetinic acid (GA) on H9c2 cells under oxygen and glucose deprivation (OGD)-induced injury, and the possible mechanisms. We found that HA+GA significantly improved pathology and morphology of the nucleus and ultrastructure of H9c2 cells under OGD as determined by Hoechst 33342 staining and transmission electron microscopy (TEM) tests. It also reduced the releases of lactate dehydrogenase (LDH), creatine kinase-myocardial band isoenzyme (CK-MB), and aspartate transaminase (AST) from the cultured supernatant of H9c2 cells, which were tested by enzyme-linked immune sorbent assay (ELISA) kits. In addition, it lessened the apoptotic rate as determined by a fluorescein isothiocyanate-annexin V/propidium iodide (FITC-AV/PI) double staining assay. It was also found that HA+GA might regulate the protein expression associated with the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Overall, the study demonstrated that HA+GA protected H9c2 cells against OGD-induced injury, and the signaling mechanism might be related to the PI3K/Akt signaling pathway.

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“…The apoptotic ratio was detected using an Annexin V/PI method (21). Briefly, the cells were treated with various concentrations of SM (0, 5, 10 or 20 µM) for 24 h, trypsinized (Gibco Life Technologies) and resuspended in 100 µl binding buffer, followed by addition of 5 µl Annexin V and PI in each tube.…”

Section: Detection Of Apoptosismentioning

confidence: 99%

Solamargine triggers hepatoma cell death through apoptosis

et al. 2015

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Abstract. Solamargine (SM), a steroidal alkaloid glycoside extracted from the traditional Chinese herb Solanum incanum, has been evidenced to inhibit the growth and induce apoptosis in a number of human cancer cell lines. In the present study, the anticancer effect of SM and underlying molecular mechanism of SM-induced apoptosis were investigated on the human hepatocellular carcinoma cells, SMMC7721 and HepG2. The proliferation effects of SM on the SMMC7721 and HepG2 cell lines were evaluated using MTT and colony formation assays. In addition, the percentage of apoptosis was measured using an Annexin V/propidium iodide staining method and the cell cycle distribution mediated by SM was analyzed using flow cytometry. The expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), caspase-3, caspase-9, proliferating cell nuclear antigen (pcna) and Ki67 proteins were examined to further demonstrate the proliferate and apoptosis effects of SM on the hepatoma cells. The results indicated that SM effectively inhibited hepatoma cell proliferation and promoted apoptosis. SM resulted in cell cycle arrest at the G 2 /M phase in the two cell lines. In addition, SM downregulated the levels of proliferation-associated (Ki67 and pcna) and anti-apoptotic (Bcl-2) proteins, and promoted the activity of apoptosis-associated proteins (Bax, caspase-3 and caspase-9). Therefore, the activation of the Bcl-2/Bax and caspase signaling pathways may be involved in the SM-induced apoptosis of hepatoma cells.

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Detection of Apoptosis Based on the Interaction between Annexin V and Phosphatidylserine

Cited by 70 publications

References 21 publications

TGF-β1 induces apoptosis of bone marrow-derived mesenchymal stem cells via regulation of mitochondrial reactive oxygen species production

TGF-β1 induces apoptosis of bone marrow-derived mesenchymal stem cells via regulation of mitochondrial reactive oxygen species production

Protective mechanisms of hypaconitine and glycyrrhetinic acid compatibility in oxygen and glucose deprivation injury

Solamargine triggers hepatoma cell death through apoptosis

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