SOCS3受miR-183-5p调控影响乳腺癌细胞的增殖、迁移、侵袭和凋亡

MiR-183-5p promotes proliferation,metastasis and inhibits apoptosis of breast cancer cells by targeting SOCS3

目的探究细胞因子信号传送阻抑物3(SOCS3)调节乳腺癌进展的分子机制。方法生物信息学方法分析TCGA数据库中乳腺癌患者的数据;体外培养乳腺癌细胞系,定量反转录聚合酶链式反应检测SOCS3和miR-183-5p的mRNA表达水平,蛋白质印迹法检测SOCS3的蛋白表达水平;对MDA-MB-231、T47D细胞进行过表达或敲低SOCS3和miR-183-5p处理,双萤光素酶报告实验检测SOCS3和miR-183-5p的靶向结合关系;CCK-8和克隆形成实验检测MDA-MB-231和T47D细胞的增殖能力;Transwell实验检测MDA-MB-231和T47D细胞的迁移和侵袭能力;流式细胞术检测MDA-MB-231和T47D细胞的凋亡率。结果SOCS3在乳腺癌组织和细胞系中低表达,而miR-183-5p在乳腺癌组织和细胞系中高表达。细胞功能实验证明了SOCS3抑制乳腺癌细胞的增殖、迁移、侵袭以及促进细胞凋亡。starBase和Targetscan数据库生物信息分析发现miR-183-5p与SOCS3有结合位点,双萤光素酶报告实验证明miR-183-5p与SOCS3具有靶向结合关系,且miR-183-5p靶向下调SOCS3的表达。细胞功能检测实验表明沉默miR-183-5p的同时沉默SOCS3,相比于单独沉默miR-183-5p,细胞的增殖、迁移和侵袭能力升高,凋亡率降低;而过表达miR-183-5p并沉默SOCS3的细胞相比于同时沉默miR-183-5p和SOCS3的增殖、迁移和侵袭能力进一步升高,凋亡率降低。结论SOCS3能抑制乳腺癌细胞的增殖、迁移、侵袭,促进其凋亡,并受miR-183-5p靶向调控。

Objective To explore the molecular mechanism of SOCS3 in regulating the progression of breast cancer.Methods Bioinformatics methods were used to analyse the data of breast cancer patients in the TCGA database;breast cancer cell lines were cultured in vitro,the mRNA expression levels of SOCS3 and miR-183-5p detected by qRT-PCR and the protein expression level of SOCS3 detected by protein blotting;MDAMB-231 and T47D cells were treated with knockdown or overexpression of SOCS3 and miR-183-5p,dual luciferase reporter assay used to test the target-binding relationship of SOCS3 and miR-183-5p;CCK-8 and clone formation assay was used to detect the proliferative ability of MDA-MB-231 and T47D cells;transwell assay used to detect the migratory and invasive ability of MDA-MB-231 and T47D cells;flow cytometry used to detect the apoptotic rate of MDA-MB-231 and T47D cells.Results SOCS3 was lowly expressed in breast cancer tissues and cell lines,whereas miR-183-5p was highly expressed.The cell function experiment proved that SOCS3 inhibited the proliferation,migration and invasion and promoted the apoptosis of breast cancer cells.Bioinformatics analysis of the starBase and Targetscan databases revealed that miR-183-5p has binding sites with SOCS3.Meanwhile,dual luciferase reporting experiments demonstrated a targeted binding relationship between miR-183-5p and SOCS3.MiR-183-5p downregulates SOCS3 expression in a targeted manner.Cell function assay experiments showed that down-regulation of miR-183-5p with simultaneous silencing of SOCS3 resulted in significantly higher proliferation,migration and invasion abilities and lower apoptosis rates compared to down-regulation of miR-183-5p alone,whereas cells overexpressing miR-183-5p and silencing of SOCS3 showed further higher proliferation,migration and invasion abilities and lower apoptosis rates compared to simultaneous silencing of miR-183-5p and SOCS3.Conclusion SOCS3 inhibits proliferation,migration,invasion and promotes apoptosis of breast cancer cells and is regulated by miR-183-5p targeting.