Effect and mechanism of down-regulating REV7 expression on the radiosensitivity of human colon cancer cell HCT116

中华放射肿瘤学杂志 Pub Date : 2020-02-15 DOI:10.3760/CMA.J.ISSN.1004-4221.2020.02.008

C. Yuan, Lu Ma, Feifei Kong

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Abstract

Objective 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 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. Key words: REV7 gene; HCT116 cell line; Radiosensitivity; Non-homologous end joining

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下调REV7表达对人结肠癌细胞HCT116放射敏感性的影响及机制

目的探讨下调REV7表达对人结肠癌细胞HCT116放射敏感性的影响及其机制。方法培养HCT116细胞，采用RNA干扰技术下调REV7的表达。将HCT116细胞分别分为空白组、阴性对照转染负RNA寡核苷酸组和REV7表达下调转染REV7寡核苷酸组。用菌落形成法测定细胞增殖情况。Western blot检测相关基因蛋白表达水平。观察细胞凋亡和非同源末端连接的水平。结果经6Gy辐照后，The REV7 siRNA组细胞集落形成率明显降低(P < 60%)。在REV7 siRNA组中，γ - h2ax和Caspase9的表达水平显著上调，KU80和XRCC4的表达水平显著下调(均P<0.05)。结论下调REV7的表达可提高人结肠癌细胞HCT116的放射敏感性。其潜在机制可能与非同源末端连接发生率较低有关。关键词:REV7基因;HCT116细胞系;辐射敏感度;非同源端连接

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中华放射肿瘤学杂志

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期刊介绍： The Chinese Journal of Radiation Oncology is a national academic journal sponsored by the Chinese Medical Association. It was founded in 1992 and the title was written by Chen Minzhang, the former Minister of Health. Its predecessor was the Chinese Journal of Radiation Oncology, which was founded in 1987. The journal is an authoritative journal in the field of radiation oncology in my country. It focuses on clinical tumor radiotherapy, tumor radiation physics, tumor radiation biology, and thermal therapy. Its main readers are middle and senior clinical doctors and scientific researchers. It is now a monthly journal with a large 16-page format and 80 pages of text. For many years, it has adhered to the principle of combining theory with practice and combining improvement with popularization. It now has columns such as monographs, head and neck tumors (monographs), chest tumors (monographs), abdominal tumors (monographs), physics, technology, biology (monographs), reviews, and investigations and research.