军事医学2024，Vol.48Issue(11)：809-814,6.DOI:10.7644/j.issn.1674-9960.2024.11.002
CDK4/6抑制剂帕博西尼通过抑制RB磷酸化防护肠上皮细胞放射损伤
CDK4/6 inhibitor palbociclib protects intestinal epithelial cells from radiation injury by inhibiting RB phosphorylation
纪奥强 1王训 2张雪文 2乔子肢 2邢爽 2申星 2余祖胤1
作者信息
- 1. 安徽医科大学生命科学学院,合肥 230032||军事科学院军事医学研究院,北京 100850
- 2. 军事科学院军事医学研究院,北京 100850
- 折叠
摘要
Abstract
Objective To investigate the protective effect and mechanism of CDK4/6 inhibitor palbociclib against radiation-induced injury in rats'small intestinal crypt epithelial cell line(IEC-6).Methods(1)Using CCK-8 colorimetric assay,colony formation assay,the PI staining method,Annexin V-FITC/PI dual staining and Western blot,the impact of palbociclib on proliferation,cell cycle dynamics,apoptosis,DNA damage,as well as phosphorylation events at Rb-780 and 795 sites in irradiated IEC-6 cells was investigated.(2)An IEC-6 cell line overexpressing Rb was established,and the effects of Rb overexpression on proliferation,cell cycle,and apoptosis in the IEC-6 cell line were assessed using the CCK-8 colorimetric assay,PI staining method,Annexin V-PE/7-AAD dual staining and Western blot.Additionally,the impact of Rb overexpression on the protective effect of palbociclib against radiation damage was verified in the Rb-overexpressing IEC-6 cell line.Results(1)Administration of palbociclib was demonstrated to enhance the proliferative capacity of IEC-6 cells following exposure to 10 Gy X-ray irradiation,increase colony formation ability post 4-8 Gy X-ray exposure while concomitantly reducing the proportion of cells in the G2/M phase after 10 Gy irradiation and diminishing apoptosis in irradiated IEC-6 cells.Protein expression analysis revealed that palbociclib inhibited phosphorylation at Rb-780 and 795 sites and attenuated the production of γ-H2AX in IEC-6 cells following irradiation.(2)After transfection of IEC-6 cells with Rb overexpression lentivirus,there was a significant increase in the total protein expression of Rb.The phosphorylation levels at Rb-780 and 795 sites were markedly elevated.Consequently,cell proliferation was accelerated,the proportion of cells in the S phase of the cell cycle increased,and the apoptotic rate significantly rose after exposure to 10 Gy X-ray irradiation.(3)Treatment with a lower dose of palbociclib(0.05 μmol/L)was found to suppress the phosphorylation of Rb at 780 and 795 sites in IEC-6 cells overexpressing Rb.Additionally,it reduced apoptosis in IEC-6 cells overexpressing Rb following exposure to 10 Gy X-ray irradiation.However,this treatment did not inhibit the phosphorylation of Rb at 780 and 795 sites in IEC-6 cells transfected with the empty vector,nor did it exert a protective effect against radiation-induced damage in these cells.Conclusion The CDK4/6 inhibitor palbociclib demonstrates significant protective effects against radiation-induced damage in rats'intestinal crypt epithelial cell line(IEC-6),with the Rb protein potentially playing a crucial role in mediating this radioprotective response.关键词
帕博西尼/电离辐射/IEC-6细胞/集落形成/凋亡/细胞周期/Rb蛋白磷酸化Key words
palbociclib/ionization radiation/IEC-6 cell/colony formation/apoptosis/cell cycle/RB phosphorylation分类
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纪奥强,王训,张雪文,乔子肢,邢爽,申星,余祖胤..CDK4/6抑制剂帕博西尼通过抑制RB磷酸化防护肠上皮细胞放射损伤[J].军事医学,2024,48(11):809-814,6.
