四溴苯三唑对人结肠癌SW480细胞凋亡的诱导作用及其机制

doi:10.13481/j.1671-587x.20170615

Abstract

Abstract: Objective:To investigate the promotion effect of 4,5,6,7-tetrabromobenzotriazole (TBB) on the apoptosis of human colon cancer SW480 cells, and to explore its possible mechanism. Methods:The human colon cancer SW480 cells at logarithmic growth phase were divided into control group (0 μmol·L^-1 TBB) and experiment group (1, 3, 10, 30, and 100 μmol·L^-1 TBB). The viability of cells was measured by MTT assay; the apoptotic rate of the SW480 cells and the level of intracellular reactive oxygen species (ROS) were analyzed by Annexin Ⅴ-FITC/PI double staining and flow cytometry. The expression levels of anti-apoptotic proteins p-Akt and Bcl-2, and pro-apoptotic proteins Bad, pro-caspase-9 and cleaved-caspase-3 were detected by Western blotting method. Results:The MTT results showed that the viabilities of SW480 cells in experiment group were decreased in a dose-dependent manner, which were lower than that in control group (P<0.05).The Annexin Ⅴ-FITC/PI double staining and flow cytometry results showed that the apoptotic rates of SW480 cells in experiment group (3, 6, 12, and 24 h)were significantly higher than that in control group (0 h) (P<0.05). The flow cytometry results showed the levels of ROS in SW480 cells after treated with TBB for 3, 6, 12 and 24 h were higher than that in 0 h group (P<0.05). The Western blotting results showed that the expression levels of anti-apoptotic proteins p-Akt and Bcl-2 in SW480 cells in experiment group (3, 6, 12, and 24 h) were decreased obviously, whereas the expression levels of the pro-apoptotic proteins Bad and cleaved-caspase-3 were increased and the expression level of pro-caspase-9 was decreased compared with those in control group (0 h) (P<0.05). Conclusion:TBB could inhibit the cell proliferation and induce the apoptosis of human colon cancer SW480 cells,and its mechanism may be related to the inhibition of the activity of Akt and the promotion of the level of intracellular ROS.

Key words: colon neoplasms, SW480 cells, reactive oxygen species, Akt signal pathway, apoptosis, 4,5,6,7-tetrabromobenzotriazole

CLC Number:

WANG Yue, LIU Chang, PIAO Xianji, ZHANG Dongyun, MENG Lingqi, WANG Hao, WANG Jiaru, LUO Yinghua, SUN Hunan, JIN Chenghao. Induction effect of tetrabromobenzotriazole on apoptosis of human colon cancer SW480 cells and its mechanism[J].Journal of Jilin University Medicine Edition, 2017, 43(06): 1148-1154.

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Induction effect of tetrabromobenzotriazole on apoptosis of human colon cancer SW480 cells and its mechanism

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