文章摘要
miR-574-3p在三阴性乳腺癌细胞紫杉醇药物敏感性中的作用
Effect of miR-574-3p on taxol sensitivity in triple-negative breast cancer cells
投稿时间:2023-07-09  
DOI:10.3969/j.issn.1000-0399.2024.03.001
中文关键词: 三阴性乳腺癌  MDA-MB-231细胞  紫杉醇  miR-574-3p  色素框同源物2  药物敏感性
英文关键词: Triple negative breast cancer  MDA-MB-231 cells  Paclitaxel  MiR-574-3p  CBX2  Drug sensitivity
基金项目:河南省医学科技攻关计划项目(编号:SBGJ202002038)
作者单位E-mail
何依珊 450052 河南郑州 郑州大学第一附属医院乳腺外科  
苏静 450052 河南郑州 郑州大学第一附属医院乳腺外科  
洪庆 450052 河南郑州 郑州大学第一附属医院乳腺外科 hqdhr@163.com 
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中文摘要:
      目的 研究miR-574-3p在三阴性乳腺癌细胞紫杉醇(PTX)药物敏感性中的作用。方法 构建稳定转染细胞株,根据细胞转染情况分为未转染组、miR-阴性对照(miR-NC)组和miR-574-3p过表达组,细胞计数试剂盒8(CCK-8)法检测不同浓度紫杉醇作用下细胞增殖情况(未转染组设置亚组:空白组和对照组),实时荧光定量聚合酶链式反应(PCR)法检测色素框同源物2(CBX2)mRNA的表达水平,Western blot法检测CBX2蛋白的表达水平,Transwell检测细胞侵袭迁移力,划痕实验检测细胞迁移能力。结果 CCK-8法检测结果显示,紫杉醇对各组三阴性乳腺癌MDA-MB-231细胞的生长抑制呈一定的浓度依赖性,但不同浓度紫杉醇(0.05、0.15、0.45、1.35、4.05、8.10 μmol/L)作用下,miR-574-3p过表达组生长抑制作用均高于对照组与miR-NC组(P<0.05);实时荧光定量PCR结果显示,2.10 μmol/L紫杉醇作用下,miR-574-3p过表达组细胞中CBX2 mRNA表达水平低于未转染组和miR-NC组(P<0.05); Western blot结果显示,2.10 μmol/L紫杉醇作用下,miR-574-3p过表达组细胞中CBX2蛋白表达水平低于未转染组和miRNC组(P<0.05); Transwell检测及划痕实验检测结果显示,2.10 μmol/L紫杉醇作用下,miR-574-3p过表达组细胞侵袭迁移能力低于未转染组和miR-NC组(P均<0.05)。结论 miR-574-3p可能通过下调CBX2增强三阴性乳腺癌细胞对紫杉醇的敏感性,抑制三阴性乳腺癌细胞的侵袭迁移能力。
英文摘要:
      Objective To study the role of miR-574-3p in the sensitivity of triple-negative breast cancer cells to paclitaxel (PTX). Methods Stable transfection cell lines were constructed and divided into the non-transfection group, miR-negative control (miR-NC) group and miR-574-3p overexpression group according to cell transfection conditions. Cell proliferation under different concentrations of paclitaxel was detected by cell counting kit 8 (CCK-8) (subgroup blank group and control group were set up in the non-transfected group), and the mRNA expression level of pigmentation box homology 2(CBX2) was detected by real-time fluorescence quantitative polymerase chain reaction (PCR). The expression level of CBX2 protein was detected by Western blotting, cell invasion and migration were evaluated by Transwell assay, and cell migration was assessed by scratch assay. Results The results of CCK-8 assay showed that paclitaxel inhibited the growth of triple negative breast cancer MDA-MB-231 cells in a dose-dependent manner in all groups, but under different concentrations of paclitaxel (0.05, 0.15, 0.45, 1.35, 4.05, 8.10 μmol/L), the growth inhibition effect of miR-574-3p overexpression group was higher than that of control group and miR-NC group (P<0.05). Real-time fluorescence quantitative PCR results showed that under the treatment of 2.10 μmol/L paclitaxel, the mRNA expression level of CBX2 in the miR-574-3p overexpression group was lower than that in the untransfected group and miR-NC group (P<0.05). Western blot results showed that under the treatment of 2.10 μmol/L paclitaxel, the expression level of CBX2 protein in the miR-574-3p overexpression group was lower than that in the untransfected group and miR-NC group (P<0.05). The results of Transwell detection and scratch test showed that under the treatment of 2.10 μmol/L paclitaxel, the cell invasion and migration ability of the miR-574-3p overexpression group was lower than that of the untransfected group and miR-NC group (P<0.05). Conclusions MiR-574-3p may enhance the sensitivity of triple-negative breast cancer cells to paclitaxel by down-regulating CBX2, and inhibit the invasion and migration ability of triple-negative breast cancer cells.
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