XRCC1 deficiency increased the DNA damage induced by γ-ray in HepG2 cell: Involvement of DSB repair and cell cycle arrest
XRCC1 deficiency increased the DNA damage induced by γ-ray in HepG2 cell: Involvement of DSB repair and cell cycle arrest
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DOI：
10.1016/j.etap.2013.04.009
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发表时间：
2013-09-01
影响因子：
4.3
通讯作者：
Zhang, Rong
中科院分区：
文献类型：
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作者：
Niu, Yujie;Zhang, Xing;Zhang, Rong
伽玛射线照射可引起不同类型细胞的DNA损伤，包括碱基损伤、单链断裂和双链断裂。DNA修复蛋白XRCC1作为BER途径的一部分，在DNA单链断裂修复中与DNA聚合酶β、DNA连接酶III和多聚ADP核糖聚合酶(PARP)形成复合体，也影响双链断裂的修复。然而，XRCC1是否参与影响HepG2细胞的辐射敏感性和DNA损伤及其可能的机制尚不清楚。因此，本研究的目的是探讨shRNA阻断XRCC1基因的表达是否会降低DNA修复，从而增强HepG2细胞对伽玛射线的敏感性。台盼蓝染色和克隆形成率测定细胞存活率。用彗星试验检测DNA损伤。流式细胞仪检测细胞凋亡率和细胞周期。荧光定量聚合酶链式反应检测DNA-PKcs和Gadd153基因的表达。我们的结果表明，取消XRCC-1可以增加HepG2细胞对伽玛射线的敏感性。这种敏感性的增强可能与DNA损伤增加和细胞周期停滞增加有关，这可能与DNA-PKcs和Gadd153mRNA表达增加有关。因此，我们的结果表明，靶向抑制XRCC1可以提高HepG2细胞的伽马射线照射敏感性。(C)2013爱思唯尔B.V.保留所有权利。
gamma-ray irradiation can induce DNA damages which include base damages, single-strand breaks and double-strand breaks in various type cells. The DNA repair protein XRCC1, as a part of the BER pathway, forms complexes with DNA polymerase beta, DNA ligase III and poly-ADP-ribose polymerase (PARP) in the repair of DNA single strand breaks and also affects the repair of double strand breaks. However, it is still not known well whether XRCC1 contributes to affect the irradiation sensitivity and DNA damage in HepG2 cell and the potential mechanism. Hence, the purpose of this study was to explore whether abrogation of XRCC1 gene expression by shRNA could reduce DNA repair and thus sensitize HepG2 cells to gamma-ray. Cell viability was measured by Trypan blue staining and cloning efficiency assay. The DNA damage was detected by Comet assay. Apoptosis and cell cycle were detected by flow cytometry. The DNA-PKcs and gadd153 mRNA expression were determined by Real-time PCR. Our results showed that abrogation of XRCC 1 could sensitize HepG2 cells to gamma-ray. This enhanced sensitivity could be attributed to the increased DNA damage and increased cell cycle arrest, which might be related with the increasing of DNA-PKcs and gadd153 mRNA expression. Therefore, our results suggested that the gamma-ray irradiation sensitivity could be increased by targeting inhibition of XRCC1 in HepG2 cell. (C) 2013 Elsevier B.V. All rights reserved.