• Title/Summary/Keyword: nonhomologous end joining

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DNA damage repair is suppressed in porcine aged oocytes

  • Lin, Tao;Sun, Ling;Lee, Jae Eun;Kim, So Yeon;Jin, Dong Il
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.984-997
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    • 2021
  • This study sought to evaluate DNA damage and repair in porcine postovulatory aged oocytes. The DNA damage response, which was assessed by H2A.X expression, increased in porcine aged oocytes over time. However, the aged oocytes exhibited a significant decrease in the expression of RAD51, which reflects the DNA damage repair capacity. Further experiments suggested that the DNA repair ability was suppressed by the downregulation of genes involved in the homologous recombination (HR) and nonhomologous end-joining (NHEJ) pathways. The expression levels of the cell cycle checkpoint genes, CHEK1 and CHEK2, were upregulated in porcine aged oocytes in response to induced DNA damage. Immunofluorescence results revealed that the expression level of H3K79me2 was significantly lower in porcine aged oocytes than in control oocytes. In addition, embryo quality was significantly reduced in aged oocytes, as assessed by measuring the cell proliferation capacity. Our results provide evidence that DNA damage is increased and the DNA repair ability is suppressed in porcine aged oocytes. These findings increase our understanding of the events that occur during postovulatory oocyte aging.

Suppression of Ku80 Correlates with Radiosensitivity and Telomere Shortening in the U2OS Telomerase-negative Osteosarcoma Cell Line

  • Hu, Liu;Wu, Qin-Qin;Wang, Wen-Bo;Jiang, Huan-Gang;Yang, Lei;Liu, Yu;Yu, Hai-Jun;Xie, Cong-Hua;Zhou, Yun-Feng;Zhou, Fu-Xiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.795-799
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    • 2013
  • Ku70/80 heterodimer is a central element in the nonhomologous end joining (NHEJ) DNA repair pathway, Ku80 playing a key role in regulating the multiple functions of Ku proteins. It has been found that the Ku80 protein located at telomeres is a major contributor to radiosensitivity in some telomerase positive human cancer cells. However, in ALT human osteosarcoma cells, the precise function in radiosensitivity and telomere maintenance is still unknown. The aim of this study was to investigate the effects of Ku80 depletion in the U2OS ALT cell line cell line. Suppression of Ku80 expression was performed using a vector-based shRNA and stable Ku80 knockdown in cells was verified by Western blotting. U2OS cells treated with shRNA-Ku80 showed lower radiobiological parameters (D0, Dq and SF2) in clonogenic assays. Furthermore, shRNA-Ku80 vector transfected cells displayed shortening of the telomere length and showed less expression of TRF2 protein. These results demonstrated that down-regulation of Ku80 can sensitize ALT cells U2OS to radiation, and this radiosensitization is related to telomere length shortening.

DNA Ligase4 as a Prognostic Marker in Nasopharyngeal Cancer Patients Treated with Radiotherapy

  • Kim, Dong Hyun;Oh, Sung Yong;Kim, So Yeon;Lee, Seul;Koh, Myeong Seok;Lee, Ji Hyun;Lee, Suee;Kim, Sung-Hyun;Park, Heon Soo;Hur, Won Joo;Jeong, Jin Sook;Ju, Mi Ha;Seol, Young Mi;Choi, Young-Jin;Chung, Joo Seop;Kim, Hyo-Jin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10985-10989
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    • 2015
  • Background: The capability for DNA double-strand breaks (DSBs) repair is crucial for inherent radiosensitivity of tumor and normal cells. We have investigated the clinicopathologic significance of DNA repair gene expression in nasopharyngeal (NP) carcinoma. Materials and Methods: A total of 65 NP cancer patients who received radiotherapy were included. The immunopositivity to Ku 70, DNA-PKcs, MRN, RAD50, XRCC4, and LIG4 were examined in all tumor tissues. Results: The patients comprised 42 males and 23 females, with a median age of 56 years (range, 18-84). The expression levels of RAD50 (0,+1,+2,+3) were 27.7%, 32.3%, 21.5%, and 18.5%. LIG4 (${\pm}$) were 43.1% and 56.9% respectively. The 5-year OS rate of patients with LIG4 (${\pm}$) were 90% and 67.9%, respectively (p=0.035). The 5-year TTP rate of patients with LIG4 (${\pm}$) were 75.9%, 55.5%, respectively (P=0.039). Conclusions: Our results suggest the possibility of predicting the radiosensitivity of NP cancer by performing immunohistochemical analysis of LIG4.