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氯硝柳胺通过下调Ku70/80的表达使鼻咽癌对放疗敏感。

Niclosamide sensitizes nasopharyngeal carcinoma to radiation by downregulating Ku70/80 expression.

作者信息

Li Jingjing, Li Haiwen, Zhan Dechao, Xiang Mei, Yang Jun, Zuo Yufang, Yu Yin, Zhou Hechao, Jiang Danxian, Luo Haiqing, Chen Zihong, Yu Zhonghua, Xu Zumin

机构信息

Cancer Center, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524000, Guangdong Province, China.

出版信息

J Cancer. 2018 Feb 6;9(4):736-744. doi: 10.7150/jca.20963. eCollection 2018.

Abstract

The aim of the present study was to investigate whether niclosamide could sensitize the nasopharyngeal carcinoma cells to radiation and further explore the underlying mechanisms. CCK-8 assay was used to determine the effect of niclosamide on the proliferation of NPC cells. Colony formation assay was used to evaluate the radiosensitizing effect of niclosamide on NPC cells. Flow cytometry analysis was used to determine the apoptosis of NPC cells induced by niclosamide. Immunofluorescent staining was used to detect the formation of γ-H2AX foci and the localization of Ku70/80 proteins in NPC cells. Real-time PCR quantification analysis was used to examine the level of Ku70/80 mRNA. DNA damage repair-related proteins were detected by western blot analysis. Our results showed that niclosamide markedly suppressed the proliferation of NPC cells. Niclosamide pretreatment followed by irradiation reduced the colony forming ability of NPC cells. Niclosamide in combination with irradiation significantly increased the apoptotic rate of NPC cells. Niclosamide significantly reduced the transcriptional level of K70/80 but not the translocation of Ku70/80 protein induced by irradiation. In conclusion, our study demonstrated that niclosamide could inhibit the growth of NPC cells and sensitize the NPC cells to radiation via suppressing the transcription of Ku70/80.

摘要

本研究的目的是探讨氯硝柳胺是否能使鼻咽癌细胞对放疗敏感,并进一步探究其潜在机制。采用CCK-8法检测氯硝柳胺对鼻咽癌细胞增殖的影响。采用集落形成试验评估氯硝柳胺对鼻咽癌细胞的放射增敏作用。采用流式细胞术分析氯硝柳胺诱导的鼻咽癌细胞凋亡情况。采用免疫荧光染色检测鼻咽癌细胞中γ-H2AX灶的形成及Ku70/80蛋白的定位。采用实时PCR定量分析检测Ku70/80 mRNA水平。通过蛋白质印迹分析检测DNA损伤修复相关蛋白。我们的结果表明,氯硝柳胺显著抑制鼻咽癌细胞的增殖。氯硝柳胺预处理后再进行照射降低了鼻咽癌细胞的集落形成能力。氯硝柳胺与照射联合使用显著提高了鼻咽癌细胞的凋亡率。氯硝柳胺显著降低K70/80的转录水平,但不影响照射诱导的Ku70/80蛋白易位。总之,我们的研究表明,氯硝柳胺可通过抑制Ku70/80的转录来抑制鼻咽癌细胞的生长并使鼻咽癌细胞对放疗敏感。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a397/5858495/4969a88cdd7a/jcav09p0736g001.jpg

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