辐射介导下HeLa细胞中Cdc25B与IER5蛋白表达的关系

doi:10.3969/j.issn.1004-616x.2017.06.001

Abstract

Abstract: OBJECTIVE:The purpose of this study was to investigate the relationship between the expression of Cdc25B and IER5 proteins in irradiated HeLa cells. METHODS:HeLa cells and IER5-knockdown Hela cells were irradiated with 4 Gy of γ rays and were harvested at 0,0.5,2,4,8,12,16,24,and 36 h after irradiation. Expression of IER5 and Cdc25B proteins was examined by Western blot. G2 cell cycle arrest was examined by flow cytometry. Expression level of Cdc25B mRNA was examined by real-time quantitative PCR. Unexposed cells were used as controls. RESULTS:The qPCR results show that within 0.5-24 h after irradiation of the normal HeLa cells the Cdc25B mRNA was lower from 0.5 h the time-pointand then increased after 2 h,the difference was statistically significant (P < 0.05);In IER5 knockdown Hela cells,Cdc25B mRNA decreased first and then increased after 0.5 h,the difference was statistically significant (all P < 0.05,except 2 h). The Western blot results show that within 0.5-24 h after irradiation of normal HeLa cells,the Cdc25B protein began to rise from the 2 h time-point,then began to decrease at 8 h,the difference was statistically significant (all P < 0.05,except 0.5 h). In IER5 knockdown Hela cells,Cdc25B began to decrease from the 0.5 h time-point,then began to rise at 8 h gradually,the difference was statistically significant (all P < 0.05,except 24 h). Compared with the control group, IER5 protein was upregulated in two cells types after their irradiation and during the timepoints mentioned above. After radiation,the level of IER5 in HeLa began to increase significantly from the 8 h time-point while IER5 knockdown Hela cells from the 0 h. The differences were statistically significant (all P < 0.05). The level of Cdc25B in HeLa began to decline significantly from 8 h,while IER5 knockdown Hela cells from 0 h. The differences were statistically significant (P < 0.05). The protein expressions of Cdc25B and IER5 were negatively correlated at the 8 h and 0 h ime-points of radiation (R₁=-0.740,R₂=-0.669,P < 0.01,respectively),while the expression of Cdc25B was decreased. The ratio of G2 cell cycle arrest was increased in the two cells lines (P < 0.05). CONCLUSION:Expression of Cdc25B protein in HeLa cells was induced by Cdc25B mRNA changes,and the expression of Cdc25B was negatively related with the expression of IER5 protein.

Key words: radiation, IER5 gene, Cdc25B, cycle arrest

CLC Number:

R811.5

ZHAO Xianzhe, LIU Xiaodan, ZHOU Pingkun, JIANG Xiaoyan, DING Kuke. Expression of Cdc25B and IER5 in irradiated HeLa cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(6): 405-410,417.

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Expression of Cdc25B and IER5 in irradiated HeLa cells

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