水飞蓟宾对结直肠癌细胞DNA损伤应答的影响  

Effect of Silibinin on DNA damage response in colorectal cancer cells

作　　者：梁国军 闫志鑫 唐钰梅 任来峰 薛耀勤[2] Liang Guojun;Yan Zhixin;Tang Yumei;Ren Laifeng;Xue Yaoqin(Third Clinical College,Shanxi University of Chinese Medicine,Jinzhong Shanxi 030619,China;Invasive Technology Department,Shanxi Province Cancer Hospital/Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences/Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan Shanxi 030000,China;Department of immunology,Shanxi Province Cancer Hospital/Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences/Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan Shanxi 030000,China)

机构地区：[1]山西中医药大学第三临床学院,山西晋中030619 [2]山西省肿瘤医院,中国医学科学院肿瘤医院山西医院,山西医科大学附属肿瘤医院介入科,山西太原030000 [3]山西省肿瘤医院,中国医学科学院肿瘤医院山西医院,山西医科大学附属肿瘤医院免疫室,山西太原030000

出　　处：《遵义医科大学学报》2023年第4期373-380,共8页Journal of Zunyi Medical University

基　　金：山西省卫健委科研项目(NO:2018066);山西省肿瘤医院肿瘤免疫学创新团队项目(NO:202003)。

摘　　要：目的探究水飞蓟宾对结直肠癌细胞DNA损伤应答的影响,并探讨其相关分子机制。方法将结直肠癌HCT116和HT29细胞分为空白对照组(DMSO)和水飞蓟宾给药组,给药组按药物浓度分为12.5、25、50、100、200μg/mL 5组。通过倒置显微镜观察给药后各组细胞的生长状态变化;采用CCK-8法检测水飞蓟宾处理后各组细胞的活力变化;采用免疫荧光法(IF)、免疫印迹法(WB)检测水飞蓟宾处理后各组细胞的DNA损伤和细胞周期相关蛋白的表达水平,包括p21、CyclinB1、Phospho-Chk2t68、Aurora A的表达水平;流式细胞术检测细胞周期分布。结果与空白组比较,水飞蓟宾对结直肠癌HCT116和HT29细胞具有明显的生长抑制作用,其抑制率随着给药浓度的增加而增强(P<0.05);与空白对照组相比,水飞蓟宾给药组DNA损伤反应蛋白γ-H2Ax表达明显升高,并且呈药物作用时间和浓度依赖性;细胞周期相关蛋白CyclinB1、Aurora A表达下调,且随着浓度的增加和作用时间的延长逐渐降低,p21、P-Chk2蛋白的表达逐渐上调,随着给药浓度的增加逐渐升高(P<0.05)。结论水飞蓟宾可以抑制结直肠癌HCT116和HT29细胞的增殖、诱导细胞DNA双链断裂损伤、细胞周期阻滞,这些结果提示水飞蓟宾可能通过DNA的损伤应答从而抑制结直肠癌的发生发展。Objective To explore the effect of Silibinin on the DNA damage response in colorectal cancer cells,and to explore the relevant molecular mechanisms.Methods Colorectal cancer HCT116 and HT29 cells were divided into blank control(DMSO)and Silibinin administration groups,including the 12.5,25,50,100 and 200μg/ml according to drug concentration.The growth status of cells in each group was observed by inverted microscope;The viability changes of cells in each group after Silibinin treatment were determined by CCK-8 assay;Immunofluorescence(IF)and western blot(WB)were used to detect the expression levels of DNA damage and cell cycle-related proteins in the cells of each group after Silibinin treatment,including p21,CyclinB1,Phospho-Chk2t68 and Aurora A;Flow cytometry was used to examine the cell cycle distribution.Results Compared with the blank group,Silibinin had a significant growth inhibition effect on colorectal cancer HCT116 and HT29 cells,and its inhibition rate was enhanced with increasing administration concentration(P<0.05);Compared with the blank control group,the expression of DNA damage response proteinγ-H2Ax was significantly increased in the Silibinin administration group and in a time and concentration dependent manner;The expression of cell cycle-related proteins Cyclin B1 and Aurora A was donhrgulated,gradually decreased with the increase of concentration and the extersion of action time,the expression of p21 and p-chk2 protein was gradually upregulated and gladually increased with the administered corcentration.(P<0.05).Conclusion Silibinin can inhibit the proliferation of colorectal cancer HCT116 and HT29 cells,induce cellular DNA double-strand break damage,and cell cycle arrest.These results suggest that Silibinin may inhibit the development of colorectal cancer through the damage response of DNA.

关 键 词：水飞蓟宾 结直肠癌 DNA损伤应答 细胞周期 

分 类 号：R735.5[医药卫生—肿瘤]