[关键词]
[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(qRT-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pcDNA-PTPN14载体,采用qRT-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、pcDNA、CyclinD1、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、pcDNA、CyclinD1、Bcl-2蛋白表达,抑制Bax、caspase-3、caspase-8蛋白表达;miR-7641与PTPN14 3'-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、pcDNA、CyclinD1、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。
[Key word]
[Abstract]
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 pcDNA-PTPN14, qRT-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, pcDNA, clinD1, 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, pcDNA, CyclinD1, Bax levels, inversely, inhibited Bcl-2, caspase-3, caspase-8 levels, miR-7641 directly targeted with PTPN14. mir-7641 and PTPN14 3' - 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, pcDNA, CyclinD1, 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.
[中图分类号]
R965
[基金项目]