Oligomycin A promotes radioresistance in HT29 colorectal cancer cells and its mechanisms.

Xiaofei Li, Ruifang Tian, Lihui Wang, Cong Xu, Hui Wu, Lan Liu, Chenghui Huang

Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 2021 Feb 28

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Radiotherapy is one of the main therapies for colorectal cancer, but radioresistance often leads to radiotherapy failure. To improve the radioresistance, we explore the effect of oligomycin A, the H+-ATP synthase inhibitor, on the sensitivity of HT29 colorectal cancer cells to irradiation and its underlying mechanisms. The effects of different concentrations of oligomycin A on the survival rate and glycolysis of HT29 colorectal cancer cells at different time points were investigated via MTT and glycolysis assay. siRNA-PFK1 was synthesized in vitro and transfected into HT29 cells. The effects of oligomycin A on radiosensitivity of HT29 colorectal cancer cells were measured via MTT and colony formation assay. Western blotting was used to detect the effect of oligomycin A on the expression of glycolytic enzyme PFK1. We compared difference between the effects of siRNA-PFK1 group and oligomycin A combined with siRNA-PFK1 group on cell survival and glycolysis. After 4 Gy X-ray irradiation, the effects of cell survival and glycolysis between the siRNA-PFK1 group and the oligomycin A combined with siRNA-PFK1 group were compared. Compared with the 0 μmol/L oligomycin A group, the cell survival rate of HT29 cells treated with 4 μmol/L oligomycin A was significantly increased (P<0.05), and the glucose uptake, the lactic acid, and the ATP production were also significantly increased (all P<0.01). After X-ray irradiation at different doses (0, 2, 4, 6, and 8 Gy), the colony formation rate and cell survival rate of the 4 μmol/L oligomycin A treated group were significantly higher than those in the 0 μmol/L oligomycin A group (both P<0.01). The sensitization enhancement ratio of oligomycin A on HT29 colorectal cancer cells was 0.4886. The expression of PFK1 in the 4 μmol/L oligomycin A group was significantly higher than that in the 0 μmol/L oligomycin A group (P<0.001). The glycolysis level, colony formation rate, and cell survival rate of the siRNA-PFK1 HT29 cells group were significantly lower than those in the 0 μmol/L oligomycin A group (all P<0.05), while the results in the 4 μmol/L oligomycin A combined with siRNA-PFK1 group were significantly higher than those in the siRNA-PFK1 group (all P<0.001). After 4 Gy X-ray irradiation, the colony formation rate and cell survival rate in the siRNA-PFK1 group were decreased compared with those in the irradiation group (P<0.01 or P<0.001), while the results of the 4 μmol/L oligomycin A combined with siRNA-PFK1 group were significantly higher than those in the siRNA-PFK1 group (both P<0.001). Oligomycin A can promote the radioresistance of HT29 colorectal cancer cells, which may be related to up-regulation of the PFK1 expression and increase of cell glycolysis.

Citation

Xiaofei Li, Ruifang Tian, Lihui Wang, Cong Xu, Hui Wu, Lan Liu, Chenghui Huang. Oligomycin A promotes radioresistance in HT29 colorectal cancer cells and its mechanisms. Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences. 2021 Feb 28;46(2):113-120