p53对核仁应激诱导卵巢癌细胞凋亡的影响  

Effect of p53 on nucleolar stress-induced apoptosis of ovarian cancer cells

作　　者：陈松斌 徐可欣 潘炫豪 胡锴克 高维浩 李松岩[1] 徐冶[1,2] CHEN Songbin;XU Kexin;PAN Xuanhao;HU Kaike;GAO Weihao;LI Song-yan;XU Ye(Laboratory of Tumor Targeted Therapy and Translational Medicine,Jilin Medical College,Jilin 132013,China;Key Laboratory of Traditional Chinese Medicine Intervention Technology for Reproductive Function Injury,Jilin Medical Col-lege,Jilin 132013,China;Department of Clinical Medicine,Jilin Medical College,Jilin 132013,China)

机构地区：[1]吉林医药学院肿瘤靶向治疗与转化医学实验室,吉林吉林132013 [2]吉林医药学院生殖功能损伤中药干预技术重点研究室,吉林吉林132013 [3]吉林医药学院临床医学院,吉林吉林132013

出　　处：《中国病理生理杂志》2023年第3期496-502,共7页Chinese Journal of Pathophysiology

基　　金：国家级大学生创新创业训练计划项目(No.201913706016);吉林省卫健委科技能力提升项目(No.2021JC048)。

摘　　要：目的:探究p53对核仁应激诱导人卵巢癌SKOV3细胞凋亡的影响。方法:体外培养SKOV3细胞,根据实验药物不同分成4个实验组:control组、BMH-21(核仁应激诱导剂)组、pifithrin-α(PFT-α;p53抑制剂)+BMH-21组和RG7112(抑制p53降解并激活p53通路)+BMH-21组。MTT法检测细胞活力;免疫荧光染色检测细胞中p53蛋白定位和表达水平,以及BMH-21诱导的核仁应激的发生;Western blot法检测蛋白表达水平;流式细胞术检测细胞凋亡率。结果:MTT结果显示,BMH-21组细胞活力比control组显著下降(P<0.01),PFT-α+BMH-21组细胞活力与BMH-21组相比无显著变化(P>0.05),而RG7112+BMH-21组比BMH-21组显著下降(P<0.01);荧光显微镜观察发现,BMH-21组p53蛋白表达水平比control组显著升高,PFT-α+BMH-21组p53蛋白表达水平比BMH-21组显著降低,而RG7112+BMH-21组比BMH-21组显著升高,且大部分入核堆积;Western blot结果显示,BMH-21组p53蛋白及线粒体途径凋亡相关蛋白表达水平比control组显著升高(P<0.01),PFT-α+BMH-21组p53蛋白表及线粒体途径凋亡相关蛋白表达水平比BMH-21组显著降低(P<0.05),而RG7112+BMH-21组比BMH-21组显著升高(P<0.01);流式细胞术结果显示,BMH-21组细胞凋亡率比control组显著升高(P<0.01),PFT-α+BMH-21组细胞凋亡率与BMH-21组相比无显著变化(P>0.05),而RG7112+BMH-21组比BMH-21组显著升高(P<0.01)。结论:BMH-21诱导人卵巢癌SKOV3细胞发生核仁应激时,抑制p53对细胞凋亡无明显影响,而激活和稳定p53会促进细胞凋亡。AIM To explore the role of p53 in the apoptosis of human ovarian cancer SKOV3 cells induced by nucleolar stress.METHODS The SKOV3 cells were divided into control group,BMH-21(nucleolar stress inducer)group,pifithrin-α(PFT-α;p53 inhibitor)+BMH-21 group and RG7112(inhibiting p53 degradation and activating p53 pathway)+BMH-21 group.Cell viability was measured by MTT assay.The localization and expression of p53 as well as the occurrence of nucleolar stress were determined by immunofluorescence.Apoptosis was evaluated by flow cytometry.RESULTS The results of MTT assay showed that the cell viabilty in BMH-21 group was significantly lower than that in control group(P<0.01),that in PFT-α+BMH-21 group was not significantly altered compared with BMH-21 group(P>0.05),and that in RG7112+BMH-21 group was significantly lower than that in BMH-21 group(P<0.01).Immunofluorescence showed that the expression of p53 protein in BMH-21 group was significantly higher than that in control group,that in PFT-α+BMH-21 group was significantly lower than that in BMH-21 group,and that in RG7112+BMH-21 group was significantly higher than that in BMH-21 group.Most p53 expression was accumulated in the nucleus.Western blotting results showed that the expression levels of p53 protein and proteins related to mitochondrial apoptosis in BMH-21 group were significantly higher than those in control group(P<0.01),those in PFT-α+BMH-21 group were significantly lower than those in BMH-21 group(P<0.05),and those in RG7112+BMH-21 group were significantly higher than those in BMH-21 group(P<0.01).Flow cytometry showed that the apoptosis rate of the cells in BMH-21 group was significantly higher than that in control group(P<0.01),that in PFT-α+BMH-21 group was not significantly altered compared with BMH-21 group(P>0.05),and that in RG7112+BMH-21 group was significantly higher than that in BMH-21 group(P<0.05).CONCLUSION Inhibition of p53 has no effect on apoptosis of human ovarian cancer SKOV3 cells after treatment with BMH-21 to induce nucleolar stres

关 键 词：卵巢癌 核仁应激 P53蛋白 细胞凋亡 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R737.31[医药卫生—基础医学]