γ-H2AX细胞周期相关性的分析

Objective: To analyze and discuss cell cycle's correlation of γ-H2AX, so as to accumulate the data for the further studies of γ-H2AX. Methods: MOLT-4 cells, and peripheral blood lymphocytes (PBLs), with or without 48 h stimulation of phytohemagglutinin (PHA), were irradiated by ultraviolet rays (UV rays). Fluorescence-labeled γ-H2AX antibody was used to detect γ-H2AX foci at the DNA double-strand breaks (DSBs) in chromatin, DNA damage was analyzed by flow cytometry, cell cycle and cell apoptosis were detected by sub-G1 peak method, the expression of y-H2AX was detected by Western blot. Results: With the progression of time, sub-G1 peak emerged apparently in the DNA histograms, and the cells of apoptosis increased gradually; with the progression of time, the increase of γ-H2AX emerged and firstly raised, then decreased; PBLs with 48 h stimulation of PHA entered apparently cell cycle, cells of S and G2/M phase emerged, and PBLs without stimulation of PHA did not enter cell cycle; Western blot showed the increase of the expression of γ-H2AX, and the increase also firstly raised, then decreased. Conclusion: γ-H2AX expressed in the cells of stationary phase and proliferative phase, and with the progression of time, the increase of γ-H2AX firstly raised, and then decreased.

Analysis of cell cycle’s correlation of γ-H2AX

Objective  To analyze and discuss cell cycle’s correlation of γ-H2AX, so as to accumulate the data for the further studies of γ-H2AX. Methods  MOLT-4 cells, and peripheral blood lymphocytes (PBLs), with or without 48 h stimulation of phytohemagglutinin (PHA), were irradiated by ultraviolet rays (UV rays). Fluorescence-labeled γ-H2AX antibody was used to detect γ-H2AX foci at the DNA double-strand breaks (DSBs) in chromatin, DNA damage was analyzed by flow cytometry, cell cycle and cell apoptosis were detected by sub-G1 peak method, the expression of γ-H2AX was detected by Western blot. Results  With the progression of time, sub-G1 peak emerged apparently in the DNA histograms, and the cells of apoptosis increased gradually; with the progression of time, the increase of γ-H2AX emerged and firstly raised, then decreased; PBLs with 48 h stimulation of PHA entered apparently cell cycle, cells of S and G2/M phase emerged, and PBLs without stimulation of PHA did not enter cell cycle; Western blot showed the increase of the expression of γ-H2AX, and the increase also firstly raised, then decreased. Conclusion  γ-H2AX expressed in the cells of stationary phase and proliferative phase, and with the progression of time, the increase of γ-H2AX firstly raised, and then decreased. Supported by grants from the National Natural Sciences Foundation of China (No. 30570908) and Clinical Key Subject Foundation of Health Ministry of China “Cell Cycle Diagnosis and Analysis in Clinical Tumor (III)”.