Journal of Shandong University (Health Sciences) ›› 2023, Vol. 61 ›› Issue (6): 22-28.doi: 10.6040/j.issn.1671-7554.0.2022.0855

• 基础医学 • Previous Articles    

Hydrogen peroxide enhances radiotherapy sensitivity of cervical cancer by regulating autophagy

REN Huixin1, ZHENG Maojin1, HAN Wencan1, WANG Chaoqun1, ZHOU Yun2, PEI Dongsheng1   

  1. 1. Laboratory of Clinical and Experimental Pathology Xuzhou Medical University, Xuzhou 221004, Jiangsu, China;
    2. Department of Radiotherapy, Xuzhou Clinical College, Xuzhou Medical University, Xuzhou 221004, Jiangsu, China
  • Published:2023-06-06

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Abstract: Objective To investigate whether hydrogen peroxide, represented as H2O2, combined with sodium hyaluronate can improve the sensitivity of cervical cancer cells to radiotherapy and to explore the underlying mechanisms. Methods The cells were divided into control group, sodium hyaluronate group, 4 Gy group, sodium hyaluronate+4 Gy group, and the expressions of autophagy related proteins were detected with Western blotting assay. Then, the cells were divided into control group, sodium hyaluronate group, 200 μmol/L H2O2 group, 500 μmol/L H2O2 group, 700 μmol/L H2O2 group and 1 000 μmol/L H2O2 group, and CCK8 experiment was used to detect the effect of H2O2 on cell viability. The cells were divided into control group and 200 μmol/L H2O2 group, and were subjected to 0, 2, 4, 6, 8 Gy radiotherapy, respectively, and the clonogenic assay was performed to assess the radiosensitizing effect of H2O2. The cells were divided into control group, 200 μmol/L H2O2 group, 4 Gy group and 200 μmol/L H2O2+4 Gy group, and the expressions of autophagy-related proteins were measured by Western blotting assay and immunofluorescence assay. Results The expressions of autophagy-related proteins LC3 II and p62 between control group and sodium hyaluronate group was not statistically significant (P>0.05). The expressions of autophagy-related proteins LC3 II and p62 between 4 Gy group and sodium hyaluronate+4 Gy group was not statistically significant(P>0.05). The results of CCK8 experiment showed that the cell proliferation rate was not statistically significant between control group and sodium hyaluronate group before and after radiotherapy(P>0.05); compared with control group, 200 μmol/L H2O2 promoted cell proliferation(P<0.05), 500 μmol/L H2O2 had no statistically significant effect on cell proliferation(P>0.05), and 700 μmol/L H2O2 and 1 000 μmol/L H2O2 inhibited cell proliferation(P<0.05). The results of clonogenesis assay showed that H2O2 increased the sensitivity of cells to radiotherapy(P<0.05). Western blotting assay and immunofluorescence assay showed that 200 μmol/L H2O2 group inhibited autophagy compared with the control group(P<0.05), and the 200 μmol/L H2O2+4 Gy group promoted autophagy compared with the 4 Gy group(P<0.05). The expressions of p-AKT and p-mTOR proteins were increased in 200 μmol/L H2O2 group compared with control group(P<0.05); while the expressions of p-AKT and p-mTOR were decreased in the 200 μmol/L H2O2+4 Gy group compared with 4 Gy group(P<0.05). Conclusion H2O2 improves radiosensitivity by regulating AKT/mTOR pathway, and then regulating autophagy.

Key words: Autophagy, Hydrogen peroxide, Sodium hyaluronate, Radiosensitivity, AKT/mTOR pathway, Cervical cancer

CLC Number: 

  • R574
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