沉默1型免疫缺陷病毒短转录诱导物连接因子-1对乳腺癌细胞多西他赛耐药性的逆转作用  

Influence of silencing factor that binds to the inducer of short transcripts of immunodeficiency virus on reversing chemoresistance to docetaxel

作　　者：徐娟[1] 付南燕[2] 傅小一[3] 崔家骏 XU Juan;FU Nan-yan;FU Xiao-yi;CUI Jia-jun(Department of Physiology and Pathophysiology,Yichun 336000,Jiangxi Province,China;Department of Pathogenic Microorganisms,Yichun 336000,Jiangxi Province,China;Department of Pathology,Yichun 336000,Jiangxi Province,China;Department of Biochemistry and Molecular Biology,Yichun College,Yichun 336000,Jiangxi Province,China)

机构地区：[1]宜春学院生理与病理生理学教研室,江西宜春336000 [2]宜春学院病原微生物教研室,江西宜春336000 [3]宜春学院病理教研室,江西宜春336000 [4]宜春学院生物化学与分子生物学教研室,江西宜春336000

出　　处：《中国临床药理学杂志》2022年第12期1334-1338,1353,共6页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(31660325)。

摘　　要：目的探讨沉默1型免疫缺陷病毒短转录诱导物连接因子-1(FBI-1)表达对乳腺癌细胞MCF-7多西他赛耐药性的逆转作用,及其可能的调控机制。方法用梯度浓度递增法建立MCF-7多西他赛耐药(MCF-7/DR)细胞株,并设置空白组、对照组和实验组。空白组不作任何转染处理;对照组转染空载体;实验组转染FBI-1特异性siRNA干扰序列载体。用实时荧光定量聚合酶链反应法检测细胞FBI-1和B细胞特异性moloney鼠白血病病毒插入位点1(Bmi-1)mRNA的表达水平,用噻唑蓝法检测MCF-7/DR细胞对多西他赛的敏感性,用流式细胞术检测10 nmol·L^(-1)多西他赛作用24 h后MCF-7/DR细胞周期和凋亡率的变化,用Western blot法检测Bmi-1蛋白的表达水平。结果多西他赛抑制空白组和实验组MCF-7/DR细胞增殖的耐药指数分别为33.08±2.33和5.24±0.35。干预24 h后,实验组、对照组和空白组的细胞凋亡率分别为(28.97±5.96)%,(7.69±1.92)%和(5.25±1.78)%,G1期细胞比例分别为(67.23±5.89)%,(58.97±5.24)%和(56.28±4.86)%,Bmi-1蛋白相对表达水平分别为0.14±0.02,0.65±0.08和0.63±0.05,实验组的上述指标与对照组和空白组比较,差异均有统计学意义(均P<0.05)。结论沉默FBI-1表达可引起MCF-7/DR细胞G1期阻滞,促使细胞凋亡,同时通过下调Bmi-1蛋白的表达,逆转MCF-7/DR细胞对多西他赛的耐药性。Objective To discuss the influence of silencing factor that binds to inducer of short transcripts-1 of human immunodeficiency virus-1(FBI-1)on chemoresistance to docetaxel of breast cancer cells MCF-7 and its possible mechanism.Methods The docetaxel-resistant MCF-7 breast cancer cell line(MCF-7/DR)was established by exposure of parental MCF-7 cells to progressively increased docetaxel concentrations,and divided into blank group,control group and experimental group.The blank group was cultured without any transfection,the control group was transfected with blank vector,and the experimental group was transfected with silencing FBI RNA sequence vector.The expression levels of FBI-1 and B-cell-specific moloney murine leukemia virus insertion site 1(Bmi-1)mRNA were measured by real-time fluorescent quantitative polymerase chain reaction.The sensitivity of MCF-7/DR cells to docetaxel was tested by methyl thiazolyl tetrazolium.Flow cytometry was used to detect the apoptosis and cell cycle of MCF-7/DR cells treated with docetaxel(10 nmol·L^(-1))for 24 h.Western blot was used to detect the expression of Bmi-1 protein.Results Resistant indexes of docetaxel on inhibiting the proliferation of MCF-7/DR cells in blank group and experimental group were 33.08±2.33 and 5.24±0.35.After treatment for 24 h,the apoptosis rates of MCF-7/DR cells in experimental group,control group and blank group were(28.97±5.96)%,(7.69±1.92)%and(5.25±1.78)%;the cell ratios in G1 phase were(67.23±5.89)%,(58.97±5.24)%and(56.28±4.86)%;the expression levels of Bmi-1 protein were 0.14±0.02,0.65±0.08 and 0.63±0.05.The above indexes in the experimental group were significantly different from those in the control group and the blank group(all P<0.05).Conclusions Silencing FBI-1 can arrest MCF-7/DR cells in G1 phase,increase the apoptosis activity and down-regulate Bmi-1 expression,therefore reversing the resistance to docetaxel of MCF-7/DR cells.

关 键 词：乳腺癌 1型免疫缺陷病毒短转录诱导物连接因子-1 B细胞特异性moloney鼠白血病病毒插入位点1 多西他赛 耐药性 

分 类 号：R97[医药卫生—药品]