AbstractObjective To evaluate the effect and mechanism of down-regulating the expression of REV7 on the radiosensitivity of human colon cancer cell HCT116. Methods HCT116 cells were cultured and the expression of REV7 was down-regulated by RNA interference technique. HCT116 cells were divided into the blank group, negative control transfected with negative RNA oligo group and REV7 expression down-regulation transfected with REV7 RNA oligo group, respectively. The cell proliferation was determined by colony formation assay. The expression levels of the proteins of relevant genes were detected by Western blot. The level of cell apoptosis and non-homologous end joining was evaluated. Results The colony formation rate was significantly reduced in THE REV7 siRNA group after 6Gy irradiation (P<0.05). The down-regulating efficiency rate of REV7 gene was>60% in the REV7 siRNA group. The expression levels of γH2AX and Caspase9 were significantly up-regulated, whereas those of KU80 and XRCC4 were remarkably down-regulated in the REV7 siRNA group (all P<0.05). Conclusions The radiosensitivity of human colon cancer cell HCT116 can be increased by down-regulating the expression of REV7. The underlying mechanism may be related to the lower incidence rate of non-homologous end joining.
Yuan Chengyang,Ma Lu,Kong Feifei et al. Effect and mechanism of down-regulating REV7 expression on the radiosensitivity of human colon cancer cell HCT116[J]. Chinese Journal of Radiation Oncology, 2020, 29(2): 115-117.
Yuan Chengyang,Ma Lu,Kong Feifei et al. Effect and mechanism of down-regulating REV7 expression on the radiosensitivity of human colon cancer cell HCT116[J]. Chinese Journal of Radiation Oncology, 2020, 29(2): 115-117.
[1] Gaotai X. REV7 counteracts DNA double-strand break resection and affects PARPinhibition[J]. Nature,2015,521(7553):541-544. DOI:10.1038/nature14328.
[2] Vera B. MAD2L2 controls DNA repair at telomeres and DNA breaks by inhibiting 5′endresection[J]. Nature,2015,521(7553):537-540. DOI:10.1038/nature14216.
[3] Junya T. REV7 is essential for DNA damage tolerance via two REV3L binding sites in mammalian DNA polymerase[J]. Nucl Acid Res,2015,43(2):1000-1011. DOI:10,1093/nar/gku1385.
[4] Sale JE. REV7/MAD2L2:the multitasking maestro emerges as a barrier to recombination[J]. EMBO J,2015,34(12):1609-1611. DOI:10.15252/embj.201591697.
[5] Yoshihiko H.3D-structured illumination microscopy reveals clustered DNA double-strand break formation in widespread γH2AX foci after high LET heavy-ion particle radiation[J]. Oncotarget,2017,8(65):109370-109381. DOI:10.18632/oncotarget.22679.
[6] 张占春. 电离辐射损伤与DNA修复基因[J]. 国际放射医学核医学杂志,2004,28(1):26-29. DOI:10.3760/cma.j.issn.1673-4114.2004.01.010.
Zhang ZC. Ionizing radiation damage and DNA repair genes[J]. Int J Radiat Med Nucl Med,2004,28(1):26-29. DOI:10.3760/cma.j.issn.1673-4114.2004.01.010.
[7] 区永聪.miRNA沉默ATM基因对HepG2在电离辐射后DNA的损伤及修复的影响[D]. 广州:南方医科大学,2011.DOI:10.7666/d.y1997587
Ou YC. Effect of microRNA silencing ATM gene on DNA damage and repair of HepG2 after ionizing radiation[D]. Guangzhou:Southern medical university,2011.DOI:10.7666/d.y1997587.
[8] Rothkamm K,Löbrich M. Evidence for a lack of DNA double-strand break repair in human cells exposed to very low x-ray doses[J]. Proc Natl Acad Sci USA,2003,100(9):5057-5062. DOI:10.1073/pans.0830918100.
[9] Yano K,Chen DJ. Live cell imaging of XLF and XRCC4 reveals a novel view of protein assembly in the non-homologous end-joining pathway[J]. Cell Cycle,2008,7(10):1321-1325. DOI:10.4161/cc.7.10.5898.
[10] Yano K,Morotomiyano K,Adachi N,et al. Molecular mechanism of protein assembly on DNA double-strand breaks in the non-homologous end-joining pathway[J]. J Radiat Res,2009,50(2):97-108. DOI:10.1269/jrr.08119.