DNA damage response following X-irradiation in oral cancer cell lines HSC3 and HSC4
DNA damage response following X-irradiation in oral cancer cell lines HSC3 and HSC4
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DOI：
10.1016/j.archoralbio.2018.02.016
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发表时间：
2018-06-01
影响因子：
3
通讯作者：
Miura, Masahiko
中科院分区：
文献类型：
--
作者：
Jiaranuchart, Sirimanas;Kaida, Atsushi;Miura, Masahiko
目的：本研究的目的是描述两个人口腔癌细胞系的DNA损伤反应后X-irradiation.Design：可视化辐射诱导的细胞周期改变，两个人口腔癌细胞系，HSC 3和HSC 4，表达荧光泛素化的细胞周期指示剂（Fucci）在这项研究中建立。从双色流式细胞术分析和Fucci荧光谱系获得照射后G2期阻滞动力学。结果：10戈伊照射后，两种细胞均出现明显的G2期阻滞，但HSC 3-Fucci细胞G2期阻滞更为明显。HSC 3-Fucci细胞的放射敏感性高于HSC 4-Fucci细胞。Fucci荧光的谱系分析显示，当在红相（G1相）中照射时，HSC 3-Fucci细胞比在红相或绿色相中照射的HSC 4-Fucci细胞表现出显著更长的绿色相（通常指示S/G2/M相，但这里反映G2停滞）。在HSC 3-Fucci细胞中，非同源末端连接在G1期期间受到轻微抑制，并且在S/G2期期间明显更可能受到损害。当G2期阻滞被检查点激酶I或Weel抑制剂废除时，只有HSC 4-Fucci细胞表现出radiosensitization.Conclusions：我们表征了照射后HSC 3-Fucci和HSC 4-Fucci细胞的DNA损伤反应，前者表现出低效的非同源末端连接，特别是在S/G2期，导致G2期阻滞增强。这些发现可能对口腔癌有临床意义。
Objective: The objective of this study was to characterize the DNA damage response in two human oral cancer cell lines following X-irradiation.Design: To visualize radiation-induced cell cycle alterations, two human oral cancer cell lines, HSC3 and HSC4, expressing fluorescent ubiquitination-based cell cycle indicator (Fucci) were established in this study. G2 arrest kinetics following irradiation were obtained from two-color flow cytometric analysis and pedigrees of Fucci fluorescence. DNA double strand break repair kinetics were obtained from immunofluorescence staining for phosphorylated histone H2AX, p53-binding protein 1, phosphorylated DNA-dependent protein kinase catalytic subunit, and breast cancer susceptibility gene 1.Results: Both cell lines showed apparent G2 arrest after 10 Gy of irradiation, but it was more enhanced in the HSC3-Fucci cells. Radiosensitivity was higher in the HSC3-Fucci cells than in HSC4-Fucci cells. Pedigree analysis of Fucci fluorescence revealed that the HSC3-Fucci cells exhibited a significantly longer green phase (normally indicating S/G2/M phases, but here reflective of G2 arrest) when irradiated in the red phase (G1 phase) than HSC4-Fucci cells irradiated in either red or green phases. Non-homologous end joining was marginally suppressed during the G1 phase and markedly more likely to be impaired during the S/G2 phases in HSC3-Fucci cells. When G2 arrest was abrogated by checkpoint kinase I or Weel inhibitors, only HSC4-Fucci cells exhibited radiosensitization.Conclusions: We characterized DNA damage response in HSC3-Fucci and HSC4-Fucci cells following irradiation and the former demonstrated inefficient non-homologous end joining, especially during the S/G2 phases, resulting in enhanced G2 arrest. These findings may have clinical implications for oral cancer.