依鲁替尼的Ibr-7衍生物通过下调磷酸化表皮生长因子受体（p-EGFR）使胰腺癌细胞对放疗增敏。

The Ibr-7 derivative of ibrutinib radiosensitizes pancreatic cancer cells by downregulating p-EGFR.

作者信息

Tan Biqin, Dong Rong, Zhang Bo, Yan Youyou, Li Qingyu, Wang Fei, Lin Nengming

机构信息

Department of Clinical Pharmacology, Key Laboratory of Clinical Cancer Pharmacology and Toxicology Research of Zhejiang Province, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, 310006 Zhejiang China.

Translational Medicine Research Center, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, 310006 Zhejiang China.

出版信息

Cancer Cell Int. 2020 Sep 17;20:458. doi: 10.1186/s12935-020-01548-6. eCollection 2020.

DOI:10.1186/s12935-020-01548-6

PMID:32963499

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7500014/

Abstract

BACKGROUND

Radiotherapy is one of the main treatments for pancreatic cancer, but radiation resistance limits its clinical application. As a result, novel therapeutic agents to improve radiosensitivity are urgently needed. This study aimed to investigate the effect of Ibr-7 (a derivative of ibrutinib) on the radiosensitivity of human pancreatic cancer cells.

METHODS

The effect of Ibr-7 on pancreatic cancer cell proliferation was detected by CCK-8 assays. Radiosensitivity was assessed by clonogenic formation assays. Cell cycle and cell apoptosis were analysed by flow cytometry. DNA damage was evaluated by immunofluorescence analysis. The expression levels of PARP, Cleaved caspase 3, p-EGFR and EGFR were determined by western blot.

RESULTS

Ibr-7 showed an anti-proliferative effect on PANC-1 and Capan2 cells in a dose- and time-dependent manner. Ibr-7 (2 μmol/L) enhanced the effect of radiation on PANC-1 and Capan2 cells. Further findings showed that this combination enhanced G2/M phase arrest and increased cell apoptosis. Additional molecular mechanism studies revealed that the expression of p-EGFR was decreased by Ibr-7 alone or in combination with radiation. Overexpression of p-EGFR reversed the cell apoptosis induced by Ibr-7 combined with radiation. Moreover, the expression of γ-H2AX was significantly decreased in the Ibr-7 plus radiation group.

CONCLUSIONS

Our study indicated the potential application of Ibr-7 as a highly effective radiosensitizer for the treatment of pancreatic cancer cells.

摘要

背景

放射治疗是胰腺癌的主要治疗方法之一，但辐射抗性限制了其临床应用。因此，迫切需要新型治疗药物来提高放射敏感性。本研究旨在探讨Ibr-7（依鲁替尼衍生物）对人胰腺癌细胞放射敏感性的影响。

方法

采用CCK-8法检测Ibr-7对胰腺癌细胞增殖的影响。通过克隆形成试验评估放射敏感性。采用流式细胞术分析细胞周期和细胞凋亡。通过免疫荧光分析评估DNA损伤。通过蛋白质印迹法测定PARP、裂解的半胱天冬酶3、p-EGFR和EGFR的表达水平。

结果

Ibr-7对PANC-1和Capan2细胞具有剂量和时间依赖性的抗增殖作用。Ibr-7（2 μmol/L）增强了辐射对PANC-1和Capan2细胞的作用。进一步研究发现，这种联合作用增强了G2/M期阻滞并增加了细胞凋亡。额外的分子机制研究表明，单独使用Ibr-7或与辐射联合使用时，p-EGFR的表达均降低。p-EGFR的过表达逆转了Ibr-7联合辐射诱导的细胞凋亡。此外，Ibr-7加辐射组中γ-H2AX的表达显著降低。