摘要
目的探讨姜黄素通过调控miR-7641/PTPN14分子轴抑制乳腺癌发展进程的分子机制。方法采用实时荧光定量PCR(q RT-PCR)检测乳腺癌患者癌组织及细胞系中miR-7641表达情况;使用Kaplan-Meier方法作乳腺癌患者生存曲线;采用不同浓度的姜黄素处理细胞,或转染miR-7641 mimic、Anti-miR-7641及pc DNA-PTPN14载体,采用q RT-PCR检测miR-7641表达情况,MTT实验及克隆形成实验检测细胞增殖能力,Transwell小室法检测细胞迁移及侵袭,western blotting检测Ki67、pc DNA、Cyclin D1、Bax、Bcl-2、caspase-3、caspase-8蛋白表达水平,采用双荧光素酶报告基因系统检测miR-7641与PTPN14靶向调控关系。结果与癌旁组织或乳腺正常上皮细胞比较,miR-7641在乳腺癌患者癌组织及乳腺癌细胞系中高表达(P<0.01、0.001),且miR-7641能够明显促进乳腺癌细胞的增殖、迁移及侵袭(P<0.05、0.01、0.001),并促进Ki67、pc DNA、Cyclin D1、Bcl-2蛋白表达,抑制Bax、caspase-3、caspase-8蛋白表达;miR-7641与PTPN143′-UTR靶向结合,姜黄素通过miR-7641/PTPN14分子轴抑制乳腺癌细胞的增殖、迁移及侵袭(P<0.01、0.001),并抑制Ki67、pc DNA、Cyclin D1、Bax蛋白表达,促进Bcl-2、caspase-3、caspase-8蛋白表达。结论姜黄素可通过下调miR-7641促进PTPN14表达,进而抑制乳腺癌细胞的增殖、迁移及侵袭。
Objective To explore the molecular mechanism of curcumin inhibiting the development of breast cancer by regulating mir-7641/ptpn14 molecular axis.Methods Detection of mir-7641 expression in breast cancer tissues and cell lines by real-time fluorescent quantitative PCR,the survival curve of breast cancer patients were analyzed using Kaplan-Meier.Breast cancer cells were treated with curcumin at different concentrations,or transfected with miR-7641 mimic,Anti-miR-7641 or pc DNA-PTPN14,q RT-PCR was used to measure miR-7641 expression,MTT assay and colony formation assay was used to determine cell viability,and Transwell assay used to performed to detect cell migration and invasion,Ki67,pc DNA,clin D1,Bax,Bcl-2,caspase-3 and caspase-8 protein level were analyzed by western blotting.Dual-luciferase report assay was performed to verify miR-7641 and PTPN14 relationship.Results miR-7641 expression was up-regulated in breast cancer tissues and cell lines(P<0.01 and 0.001).miR-7641 promoted cell proliferation,migration and invasion(P<0.05,0.01 and 0.001),up-regulated Ki67,pc DNA,Cyclin D1,Bax levels,inversely,inhibited Bcl-2,caspase-3,caspase-8 levels,miR-7641 directly targeted with PTPN14.mir-7641 and PTPN143′-UTR targeted binding,curcumin through mir-7641/PTPN14 molecular axis inhibit the proliferation,migration and invasion of breast cancer cells(P<0.01,0.001),and inhibit the expression of Ki67,pc DNA,Cyclin D1,Bax protein,promote the expression of Bcl-2,Caspase-3,Caspase-8 protein.Conclusion Curcumin inhibited breast cancer cells proliferation,migration and invasion by down-regulating miR-7641 to increasing PTPN14 expression.
作者
孟亚
张晓雯
刘玉国
MENG Ya;ZHANG Xiaowen;LIU Yuguo(College of Medicine and Life Sciences,Shandong Academy of Medical Sciences,University of Jinan,Jinan 250200,China;Department of Pharmacy,Shandong Cancer Hospital,Shandong University,Jinan 250117,China)
出处
《药物评价研究》
CAS
2020年第1期42-51,共10页
Drug Evaluation Research
