Involvement of APOBEC3B in mutation induction by irradiation.
Involvement of APOBEC3B in mutation induction by irradiation.
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DOI：
10.1093/jrr/rraa069
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发表时间：
2020-11-16
影响因子：
2
通讯作者：
Yamamoto F
中科院分区：
文献类型：
--
作者：
Saito Y;Miura H;Takahashi N;Kuwahara Y;Yamamoto Y;Fukumoto M;Yamamoto F
为了更好地了解辐射带来的癌症风险以及对辐射治疗耐药的癌细胞的发展，我们研究了癌症风险因子 APOBEC3B 在辐射诱导突变的产生中的参与情况。通过实时定量 PCR 测定了三种人类癌细胞系中 APOBEC3B 响应辐射的表达。使用次黄嘌呤-鸟嘌呤磷酸核糖转移酶 (HPRT) 突变测定，比较了 APOBEC3B 缺陷型人肝细胞癌 (HepG2) 细胞 [使用 CRISPR-Cas9 基因组编辑敲除 (KO) APOBEC3B] 和亲本细胞系之间由辐射引起的 HPRT 基因突变。然后，单独培养HPRT突变细胞以进行HPRT外显子的PCR和DNA测序。 X 射线照射诱导 HepG2、人宫颈癌上皮癌 (HeLa) 和人口腔鳞状细胞癌 (SAS) 细胞中 APOBEC3B 表达。 APOBEC3B 的强制表达增加了自发突变。相比之下，APOBEC3B KO不仅降低了自发突变率，而且强烈抑制了亲代细胞系辐射后突变频率的增加。虽然 APOBEC3B 在细胞核中的强制表达会导致 DNA 损伤，但较高水平的 APOBEC3B 往往会减少 APOBEC3B 诱导的 γ-H2AX 病灶形成（DNA 损伤修复的一种测量方法）。此外，在辐射前后，稳定表达APOBEC3B的细胞中γ-H2AX焦点的数量并不比对照高很多，表明DNA修复途径可能被激活。这项研究表明，辐射诱导 HepG2、HeLa 和 SAS 细胞中 APOBEC3B 持续表达，并且 APOBEC3B 增强辐射诱导的部分缺失。
To better understand the cancer risk posed by radiation and the development of radiation therapy resistant cancer cells, we investigated the involvement of the cancer risk factor, APOBEC3B, in the generation of radiation-induced mutations. Expression of APOBEC3B in response to irradiation was determined in three human cancer cell lines by real-time quantitative PCR. Using the hypoxanthine-guanine phosphoribosyl transferase (HPRT) mutation assay, mutations in the HPRT gene caused by irradiation were compared between APOBEC3B-deficient human hepatocellular carcinoma (HepG2) cells [APOBEC3B knocked out (KO) using CRISPR-Cas9 genome editing] and the parent cell line. Then, HPRT-mutated cells were individually cultured to perform PCR and DNA sequencing of HPRT exons. X-Irradiation induced APOBEC3B expression in HepG2, human cervical cancer epithelial carcinoma (HeLa) and human oral squamous cell carcinoma (SAS) cells. Forced expression of APOBEC3B increased spontaneous mutations. By contrast, APOBEC3B KO not only decreased the spontaneous mutation rate, but also strongly suppressed the increase in mutation frequency after irradiation in the parent cell line. Although forced expression of APOBEC3B in the nucleus caused DNA damage, higher levels of APOBEC3B tended to reduce APOBEC3B-induced γ-H2AX foci formation (a measure of DNA damage repair). Further, the number of γ-H2AX foci in cells stably expressing APOBEC3B was not much higher than that in controls before and after irradiation, suggesting that a DNA repair pathway may be activated. This study demonstrates that irradiation induces sustained expression of APOBEC3B in HepG2, HeLa and SAS cells, and that APOBEC3B enhances radiation-induced partial deletions.
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影响因子：
30.8
作者：
Burns, Michael B.;Temiz, Nuri A.;Harris, Reuben S.
通讯作者：
Harris, Reuben S.
影响因子：
3.7
作者：
Akre MK;Starrett GJ;Quist JS;Temiz NA;Carpenter MA;Tutt AN;Grigoriadis A;Harris RS
通讯作者：
Harris RS
影响因子：
13.6
作者：
Law EK;Sieuwerts AM;LaPara K;Leonard B;Starrett GJ;Molan AM;Temiz NA;Vogel RI;Meijer-van Gelder ME;Sweep FC;Span PN;Foekens JA;Martens JW;Yee D;Harris RS
通讯作者：
Harris RS
影响因子：
16.6
作者：
Adewoye, Adeolu B.;Lindsay, Sarah J.;Dubrova, Yuri E.;Hurles, Matthew E.
通讯作者：
Hurles, Matthew E.
影响因子：
2.7
作者：
Shimizu A;Fujimori H;Minakawa Y;Matsuno Y;Hyodo M;Murakami Y;Yoshioka KI
通讯作者：
Yoshioka KI