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. 2015 Nov 23:21:3629-33.
doi: 10.12659/msm.894006.

Impact of Mitochondria-Mediated Apoptosis in U251 Cell Cycle Arrest in G1 Stage and Caspase Activation

Lei Zhang  1 , Peng Liang  1 , Rui Zhang  2
Affiliations

Impact of Mitochondria-Mediated Apoptosis in U251 Cell Cycle Arrest in G1 Stage and Caspase Activation

Lei Zhang et al. Med Sci Monit. .

Abstract

BACKGROUND Most mitochondria-mediated apoptosis has some relevance to the cell cycle, but there is still a lack of investigations about U251 cell cycle in human brain glioma cells. In this study, we aimed to clarify the correlation of mitochondria-mediated apoptosis with the U251 cell cycle and its influence on apoptosis, through observing the impact of mitochondria-mediated apoptosis in U251cell specificity cycle arrest and Caspase activation. MATERIAL AND METHODS AnnexinV/PI and API were used to label the brain glioma cells for flow cytometry analysis of U251 cell apoptosis and cell cycle. RT-PCR and Western blot were performed to detect Caspase-3 and Caspase-9 activation. RESULTS Peripheral blood in stationary phase is not sensitive to apoptosis induction, but U251 cells have obvious apoptosis. Mitochondria-mediated apoptosis mainly occurs in the G1 phase of the cell cycle. Caspase-3 and Caspase-9 mRNAs and proteins expression increased significantly after the cells were treated by mitochondrial apoptosis-related gene Bax induction. CONCLUSIONS Mitochondria-mediated apoptosis is related to the U251 cell cycle with specific G1 stage arrest. Caspase activation occurs in the process of cell apoptosis.

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Figures

Figure 1
Figure 1
Cell cycle of human brain glioma cell line U251 after stimulation by tPA. * P<0.05 Compared with G2~M cells in the experimental group.
Figure 2
Figure 2
Annexin V/PI test cell apoptosis. (A) Mitochondria-mediated human brain glioma cell line U251 cell apoptosis scatter plot; (B) Mitochondria-mediated tPA stimulated human brain glioma cell line U251 cell apoptosis scatter plot.
Figure 3
Figure 3
API detect cell cycle specificity.
Figure 4
Figure 4
Cyclin E/DNA double staining flow cytometry analysis on cell cycle. (A) Control; (B) tPA; (C) Bax+tPA.
Figure 5
Figure 5
Caspase-3 and −9 protein expression.
See this image and copyright information in PMC

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