帕博西尼诱导 DNA 损伤并抑制 DNA 修复，从而诱导口腔鳞状细胞癌发生细胞衰老和凋亡。

Palbociclib induces DNA damage and inhibits DNA repair to induce cellular senescence and apoptosis in oral squamous cell carcinoma.

机构信息

Tissue Bank, Chang Gung Memorial Hospital, Tao-Yuan 33305, Taiwan; Graduate Institute of Health Industry Technology and Research Center for Chinese Herbal Medicine, College of Human Ecology, Chang Gung University of Science and Technology, Tao-Yuan 33303, Taiwan; Liver Research Center, Department of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Tao-Yuan 33305, Taiwan.

Tissue Bank, Chang Gung Memorial Hospital, Tao-Yuan 33305, Taiwan; Graduate Institute of Natural Products, Chang Gung University, Tao-Yuan 33303, Taiwan.

出版信息

J Formos Med Assoc. 2021 Sep;120(9):1695-1705. doi: 10.1016/j.jfma.2020.12.009. Epub 2020 Dec 18.

DOI:10.1016/j.jfma.2020.12.009

PMID:33342707

Abstract

BACKGROUND/PURPOSE: Palbociclib is an FDA-approved cyclin-dependent kinase (CDK) 4/6 inhibitor that has been clinically proven to be effective in breast cancer. However, its use in oral cancer is not well researched. In this study, we investigated the inhibitory activity of palbociclib against oral squamous cell carcinoma (OSCC) cells and explored the mechanism of inhibition.

METHODS

The effects of palbociclib on the cytotoxicity of OSCC cells were determined by MTT and colony formation assays. β-Galactosidase staining and cell-cycle analysis were used to determine palbociclib-induced cellular senescence and apoptosis of OSCC cells. Wound healing and transwell assays were performed to assess the effects of palbociclib treatment on migration and invasion ability of OSCC cells. Whole transcriptome sequencing was conducted to show the relationship between DNA damage repair of OSCC cells and palbociclib treatment. Palbociclib-induced DNA damage and repair capacity of OSCC cells were confirmed by comet assay and immunofluorescence confocal microscopy. Western blotting was used to verify the palbociclib-mediated changes in the CDK/pRB/c-Myc/CDC25A pathway. Finally, in vitro findings were tested in a mouse xenograft model.

RESULTS

Our results showed that palbociclib can significantly inhibit the growth, migration, and invasive ability of OSCC cells and can accelerate cellular senescence and apoptosis. We found that palbociclib induced DNA damage and p21 expression through the p53-independent pathway, thereby downregulating c-Myc and CDC25A expression to inhibit cell cycle progression. In addition, palbociclib downregulated RAD51 expression to inhibit DNA damage repair ability of OSCC cell.

CONCLUSION

Palbociclib was found to have anti-oral squamous cell carcinoma activity and to simultaneously induce DNA damage and inhibit its repair, and to accelerated cellular senescence and apoptosis.

摘要

背景/目的：帕博西尼（Palbociclib）是一种已被 FDA 批准的细胞周期蛋白依赖性激酶（CDK）4/6 抑制剂，已被临床证明对乳腺癌有效。然而，其在口腔癌中的应用尚未得到充分研究。在这项研究中，我们研究了帕博西尼对口腔鳞状细胞癌（OSCC）细胞的抑制活性，并探讨了其抑制机制。

方法

通过 MTT 和集落形成实验确定帕博西尼对 OSCC 细胞的细胞毒性作用。β-半乳糖苷酶染色和细胞周期分析用于确定帕博西尼诱导 OSCC 细胞的细胞衰老和凋亡。划痕愈合和 Transwell 实验用于评估帕博西尼处理对 OSCC 细胞迁移和侵袭能力的影响。全转录组测序用于显示 OSCC 细胞的 DNA 损伤修复与帕博西尼处理之间的关系。通过彗星实验和免疫荧光共聚焦显微镜确认帕博西尼诱导的 OSCC 细胞 DNA 损伤和修复能力。Western blot 用于验证帕博西尼介导的 CDK/pRB/c-Myc/CDC25A 通路变化。最后，在体外实验中进行了小鼠异种移植模型的检测。

结果

我们的结果表明，帕博西尼可以显著抑制 OSCC 细胞的生长、迁移和侵袭能力，并加速细胞衰老和凋亡。我们发现，帕博西尼通过 p53 非依赖性途径诱导 DNA 损伤和 p21 表达，从而下调 c-Myc 和 CDC25A 表达以抑制细胞周期进程。此外，帕博西尼下调 RAD51 表达以抑制 OSCC 细胞的 DNA 损伤修复能力。