[關(guān)鍵詞]
[摘要]
目的 探討姜黃素通過調(diào)控miR-7641/PTPN14分子軸抑制乳腺癌發(fā)展進(jìn)程的分子機制。方法 采用實時熒光定量PCR(qRT-PCR)檢測乳腺癌患者癌組織及細(xì)胞系中miR-7641表達(dá)情況;使用Kaplan-Meier方法作乳腺癌患者生存曲線;采用不同濃度的姜黃素處理細(xì)胞,或轉(zhuǎn)染miR-7641 mimic、Anti-miR-7641及pcDNA-PTPN14載體,采用qRT-PCR檢測miR-7641表達(dá)情況,MTT實驗及克隆形成實驗檢測細(xì)胞增殖能力,Transwell小室法檢測細(xì)胞遷移及侵襲,western blotting檢測Ki67、pcDNA、CyclinD1、Bax、Bcl-2、caspase-3、caspase-8蛋白表達(dá)水平,采用雙熒光素酶報告基因系統(tǒng)檢測miR-7641與PTPN14靶向調(diào)控關(guān)系。結(jié)果 與癌旁組織或乳腺正常上皮細(xì)胞比較,miR-7641在乳腺癌患者癌組織及乳腺癌細(xì)胞系中高表達(dá)(P<0.01、0.001),且miR-7641能夠明顯促進(jìn)乳腺癌細(xì)胞的增殖、遷移及侵襲(P<0.05、0.01、0.001),并促進(jìn)Ki67、pcDNA、CyclinD1、Bcl-2蛋白表達(dá),抑制Bax、caspase-3、caspase-8蛋白表達(dá);miR-7641與PTPN14 3'-UTR靶向結(jié)合,姜黃素通過miR-7641/PTPN14分子軸抑制乳腺癌細(xì)胞的增殖、遷移及侵襲(P<0.01、0.001),并抑制Ki67、pcDNA、CyclinD1、Bax蛋白表達(dá),促進(jìn)Bcl-2、caspase-3、caspase-8蛋白表達(dá)。結(jié)論 姜黃素可通過下調(diào)miR-7641促進(jìn)PTPN14表達(dá),進(jìn)而抑制乳腺癌細(xì)胞的增殖、遷移及侵襲。
[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.
[中圖分類號]
R965
[基金項目]