Silencing Artemis Enhances Colorectal Cancer Cell Sensitivity to DNA-Damaging Agents.
Silencing Artemis Enhances Colorectal Cancer Cell Sensitivity to DNA-Damaging Agents.
复制标题
沉默 Artemis 可增强结直肠癌细胞对 DNA 损伤剂的敏感性。
DOI：
10.3727/096504018x15179694020751
复制
发表时间：
2018-12-27
影响因子：
3.1
通讯作者：
Sun X
中科院分区：
文献类型：
--
作者：
Liu H;Wang X;Huang A;Gao H;Sun Y;Jiang T;Shi L;Wu X;Dong Q;Sun X
Artemis是非同源末端连接（nonhomologous end joining，NHEJ）的关键蛋白，NHEJ是哺乳动物细胞修复IR诱导的DSB的主要途径。然而，Artemis在肿瘤中的表达以及沉默Artemis对肿瘤对辐射敏感性的影响尚未得到充分研究。在这项研究中，我们研究了Artemis的表达水平如何影响结直肠癌细胞放疗和化疗的治疗结果。首先，我们发现Artemis在一些人直肠癌样本中的表达很强，使用免疫组织化学染色高于邻近的正常组织。然后我们使用慢病毒介导的siRNA敲低了人结直肠癌细胞系（RKO）中的Artemis基因。与对照组相比，Artemis基因敲除的RKO细胞对博莱霉素、依托泊苷、喜树碱和IR的敏感性显著增加; γ-H2 AX焦点法检测发现DNA修复动力学延迟;凋亡检测和Western blot检测发现Artemis基因敲除的RKO细胞凋亡明显增加。我们还发现p53/p21信号通路可能参与了细胞凋亡过程。总之，我们的研究表明，操纵Artemis可以增强结直肠癌细胞对DNA损伤剂的敏感性。因此，Artemis可以作为直肠癌治疗的治疗靶点。
Artemis is a key protein of NHEJ (nonhomologous end joining), which is the major pathway for the repair of IR-induced DSBs in mammalian cells. However, the expression of Artemis in tumors and the influence of silencing Artemis on tumor sensitivity to radiation have not been investigated fully. In this study, we investigated how the expression levels of Artemis may affect the treatment outcome of radiotherapy and chemotherapy in colorectal cancer cells. First, we found that the expression of Artemis is strong in some human rectal cancer samples, being higher than in adjacent normal tissues using immunohistochemical staining. We then knocked down Artemis gene in a human colorectal cancer cell line (RKO) using lentivirus-mediated siRNAs. Compared to the control RKO cells, the Artemis knockdown cells showed significantly increased sensitivity to bleomycin, etoposide, camptothecin, and IR. Induced by DNA-damaging agents, delayed DNA repair kinetics was found by the γ-H2AX foci assay, and a significantly increased cell apoptosis occurred in the Artemis knockdown RKO cells through apoptosis detection methods and Western blot. We also found that the p53/p21 signaling pathway may be involved in the apoptosis process. Taken together, our study indicates that manipulating Artemis can enhance colorectal cancer cell sensitivity to DNA-damaging agents. Therefore, Artemis can serve as a therapeutic target in rectal cancer therapy.
影响因子：
14.9
作者：
Mohapatra S;Kawahara M;Khan IS;Yannone SM;Povirk LF
通讯作者：
Povirk LF