Tian Jing, Kong Enqi, Wang Xiangyu, Xie Zhaoguang, Chang Cherry Yin-Yi, Sheu Jim Jinn-Chyuan, Hao Quan, Sun Li
Department of Gynecological Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin, People's Republic of China, Key Laboratory of Cancer Prevention and Therapy, Tianjin, Tianjin's Clinical Research Center for Cancer, Tianjin 300060, People's Republic of China.
School of Medicine and Life Sciences, University of Jinan-Shandong Academy of Medical Sciences, Jinan 250200, People's Republic of China.
Onco Targets Ther. 2020 Apr 9;13:3061-3071. doi: 10.2147/OTT.S246632. eCollection 2020.
Remodeling and spacing factor-1 (RSF-1) is an identified tumor biomarker that is overexpressed in a variety of human cancers, but its effect on radiotherapy remains unclear. In this study, we aimed to explore the effect of RSF-1 siRNA on sensitizing cervical cancer cells to radiation and its underlying mechanism.
The mRNA and protein expression of RSF-1 in tissue and cells were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Cell counting kit-8 (CCK-8) and colony formation assay were used to examine cell proliferation. Flow cytometry was used to analyzed the cell cycle and cell apoptosis. DNA damage was examined by the comet assay. ATM, ATR, CHK1, CHK2, H2AX, γH2AX and phosphorylated ATM, ATR, CHK1 and CHK2 were detected by Western blotting. γH2AX foci were demonstrated by immunofluorescence staining.
RSF-1 was upregulated in cervical cancer tissue and decreased after effective treatment. RSF-1 siRNA in combination with radiation suppressed cell viability, redistributed cell cycles and also induced cell apoptosis in HeLa and SiHa cell lines. Further, knockdown of RSF-1 induced DNA damage by attenuating DNA repair capability, thereby sensitizing cervical cancer cells to radiation.
These data demonstrate that RSF-1 siRNA enhanced the sensitivity of radiotherapy, and targeting RSF-1 may be a promising approach for the development of novel radiosensitizing agents for the treatment of cervical cancer.
重塑与间距因子-1（RSF-1）是一种已确定的肿瘤生物标志物，在多种人类癌症中过表达，但其对放疗的影响仍不清楚。在本研究中，我们旨在探讨RSF-1小干扰RNA（siRNA）对宫颈癌细胞放疗增敏的作用及其潜在机制。
采用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测组织和细胞中RSF-1的mRNA和蛋白质表达。使用细胞计数试剂盒-8（CCK-8）和集落形成试验检测细胞增殖。采用流式细胞术分析细胞周期和细胞凋亡。通过彗星试验检测DNA损伤。通过蛋白质免疫印迹法检测ATM、ATR、CHK1、CHK2、H2AX、γH2AX以及磷酸化的ATM、ATR、CHK1和CHK2。通过免疫荧光染色显示γH2AX焦点。
RSF-1在宫颈癌组织中上调，有效治疗后降低。RSF-1 siRNA联合放疗可抑制HeLa和SiHa细胞系的细胞活力，重新分布细胞周期并诱导细胞凋亡。此外，敲低RSF-1通过减弱DNA修复能力诱导DNA损伤，从而使宫颈癌细胞对放疗敏感。
这些数据表明，RSF-1 siRNA增强了放疗敏感性，靶向RSF-1可能是开发新型宫颈癌放疗增敏剂的一种有前景的方法。
