期刊文献+

miR-339-5p调控EMT影响乳腺癌细胞化疗耐药的机制

Mechanism of miR-339-5p influencing chemotherapy resistance of breast cancer cells via regulating EMT
下载PDF
导出
摘要 目的分析miR-339-5p调控上皮细胞-间充质转化(EMT)影响乳腺癌细胞化疗耐药的机制。方法于2023年2月至2024年2月体外培养正常乳腺上皮细胞系(MCF10A)及乳腺癌细胞系MCF-7,构建乳腺癌紫杉醇耐药细胞系MCF-7/紫杉醇,将MCF-7/紫杉醇细胞分别转染miR-339-5p模拟物(miR-339-5p mimics组)、miR-339-5p模拟物阴性对照(miR-NC组)、miR-339-5p抑制剂(miR-339-5p inhibitor组)及miR-339-5p抑制剂对照(NC组)。采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)对miR-339-5p水平予以检测,以噻唑蓝(MTT)法测定细胞增殖抑制率,经流式细胞仪测定细胞凋亡情况,采用Western Blot法检测EMT相关蛋白[E-cadhesin(E-cad)、Vimentin(Vim)]的表达。采用独立样本t检验、ONE-WAY ANOVA分析及LSD-t检验对所获得的数据进行统计学分析。结果miR-339-5p在MCF-7细胞的表达水平低于MCF10A细胞[(0.36±0.07)比(0.73±0.08),P<0.05];转染48 h后,miR-339-5p mimics组细胞增殖抑制率、细胞凋亡率高于miR-NC组[(45.86±4.92)%比(20.44±2.16)%、(16.54±1.67)%比(4.23±0.45)%,均P<0.05],miR-339-5p inhibitor组细胞增殖抑制率、细胞凋亡率低于NC组[(9.74±1.05)%比(17.18±1.84)%、(2.28±0.46)%比(5.43±0.59)%,均P<0.05];miR-339-5p mimics组E-cad水平高于miR-NC组,Vim低于miR-NC组[(0.78±0.07)比(0.42±0.05)、(0.42±0.05)比(0.61±0.07),均P<0.05],miR-339-5p inhibitor组E-cad低于NC组,Vim高于NC组[(0.23±0.04)比(0.34±0.05)、(0.84±0.09)比(0.69±0.08),均P<0.05]。结论miR-339-5p可降低乳腺癌细胞化疗耐药性,抑制细胞增殖并促进细胞凋亡,其机制可能是通过调控EMT相关蛋白E-cad、Vim而抑制EMT。 Objective To analyze the mechanism of miR-339-5p influencing chemotherapy resistance of breast cancer cells via regulating epithelial mesenchymal transformation(EMT).Methods From February 2023 to February 2024,MCF10A mammary epithelial cells and MCF-7 breast cancer cell lines were cultured in vitro.The paclitaxel resistant breast cancer cell line(MCF-7/paclitaxel)was constructed.The MCF-7/paclitaxel cells were divided into a miR-339-5p mimics group,a miR-339-5p mimetic negative control group(miR-NC group),a miR-339-5p inhibitor group,and a miR-339-5p inhibitor control group(NC group).The real-time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression level of miR-339-5p.The methyl thiazolyl tetrazolium(MTT)assay was used to determine the inhibition rate of cell proliferation.The flow cytometry was used to determine cell apoptosis.The expression of EMT related proteins[E-cadherin(E-cad)and Vimentin(Vim)]was detected using the Western blot method.The independent sample t test,one-way ANOVA,and LSD-t test were used for the statistical analysis of the obtained data.Results The expression level of miR-339-5p in the MCF-7 cells was lower than that in the MCF10A cells[(0.36±0.07)vs.(0.73±0.08);P<0.05].Forty-eight h after transfection,the proliferation inhibition rate and apoptosis rate in the miR-339-5p mimics group were higher than those in the miR-NC group[(45.86±4.92)%vs.(20.44±2.16)%and(16.54±1.67)%vs.(4.23±0.45)%;both P<0.05].The cell proliferation inhibition rate and apoptosis rate of the miR-339-5p inhibitor group were lower than those of the NC group[(9.74±1.05)%vs.(17.18±1.84)%and(2.28±0.46)%vs.(5.43±0.59)%,both P<0.05].The E-cad level in the miR-339-5p mimics group was higher than that in the miR-NC group[(0.78±0.07)vs.(0.42±0.05);P<0.05];the Vim level in the miR-339-5p mimics group was lower than that in the miR-NC group[(0.42±0.05)vs.(0.61±0.07);P<0.05].The E-cad level in the miR-339-5p inhibitor group was lower than that in the NC group[(0.23±0.04)vs.(0.34±0.05);P<0.05];the Vim level in the miR-339-5p inhibitor group was higher than that in the NC group[(0.84±0.09)vs.(0.69±0.08);P<0.05].Conclusions MiR-339-5p can reduce chemotherapy resistance of breast cancer cells,inhibit cell proliferation,and promote cell apoptosis.The mechanism may be that it inhibits EMT regulating EMT related proteins--E-cad and Vim.
作者 李志路 乔喜婷 焦婉 王小娟 司小敏 Li Zhilu;Qiao Xiting;Jiao Wan;Wang Xiaojuan;Si Xiaomin(First ward and Anning Ward,Cancer Center,Xianyang Central Hospital,Xianyang 712000,China)
出处 《国际医药卫生导报》 2024年第19期3250-3254,共5页 International Medicine and Health Guidance News
基金 吴阶平医学基金会临床科研专项资助基金(320.6750.2022-18-45)。
关键词 乳腺癌 miR-339-5p 上皮细胞-间充质转化 化疗耐药 机制 Breast cancer MiR-339-5p Epithelial mesenchymal transformation Chemotherapy resistance Mechanism
  • 相关文献

参考文献13

二级参考文献126

  • 1Carthew R W, Sontheimer E J. Origins and mechanisms of miR- NAs and siRNAs [J]. Cell, 2009,136(4) :642 -55.
  • 2Lu J, Getz G, Miska E A, et al. MicroRNA expression profiles classify human cancers[ J ]. Nature, 2005,435 (7043) :834 -8.
  • 3Bartel D P. MicroRNAs : genomics, biogenesis, mechanism, and function[J]. Cell, 2004,116(2) :2Sl -97.
  • 4Wu Z S, Wu Q, Wang C Q, et al. MiR-339-Sp inhibits breast cancer cell migration and invasion in vitro and may be a potential biomarker for breast cancer prognosis cancer [ J ]. BMC Cancer, 2010,10:542.
  • 5Wu Z S, Wang C Q, Xiang R, et al. Loss of miR-133a expression associated with poor survival of breast cancer and restoration of miR-133a expression inhibited breast cancer cell growth and inva- sion[J]. BMC Cancer, 2012,12:51.
  • 6Lander E S, Linton L M, Birren B, et al. Initial sequencing and analysis of the human genome [ J ]. Nature, 2001,409 (6822) : 860 - 921.
  • 7Calin G A, Dumitru C D, Shimizu M, et al. Frequent deletions and down-regulation of microRNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia[ J ]. Proc Natl Acad Sci USA, 2002,99 : 15524 - 9.
  • 8Hurst D R, Edmonds M D, Welch D R, et al. Metastamir: the field of metastasis-regulatory microRNA is spreading [ J ]. Cancer Res, 2009,69 ( 19 ) :7495 - 8.
  • 9Tie J, Pan Y, Zhao L, et al. MiR-218 inhibits invasion and me- tastasis of gastric cancer by targeting the Robol receptor [ J . PLoS Genet, 2010,6(3) :e1000879.
  • 10Parekh S, Prive G, Melnick A, et al. Therapeutic targeting of the BCL-6 oncogene for diffuse large B-cell lymphomas [ J ]. Leuk Lymphoma, 2008,49(5 ) :874 - 82.

共引文献50

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部