为了分析miR-484在乳腺癌组织和细胞中的表达情况,研究miR-484在乳腺癌细胞增殖、转移和自噬过程中的作用机制,首先,采用GEO数据库分析乳腺癌组织中差异表达miRNA谱,并用实时荧光定量PCR在临床乳腺癌组织及其配对的癌旁组织中检测miR-484的表达情况;其次,利用miR-484模拟物、抑制剂检测miR-484对MCF-7乳腺癌细胞增殖、转移和自噬能力的影响;再次,预测miR-484的调控基因并进行验证,同时构建 Sorbin 和 SH3 结构域包含蛋白 2(Sorbin and SH3 domain-containing protein 2,SORBS2)过表达载体,检测SORBS2对乳腺癌细胞增殖、转移和自噬能力的影响;最后,用Western-blot分析miR-484调控下MCF-7细胞中丝裂原活化的胞外信号调节激酶(mitogen-activated extracellular signal-regulated kinase,MEK)/p-MEK 和胞外信号调节激酶(extracellular signal-regulated kinase,ERK)/p-ERK的蛋白质含量变化.结果显示,miR-484在乳腺癌组织及细胞中高表达,具有提高乳腺癌细胞增殖、迁移和侵袭能力以及降低乳腺癌细胞自噬能力的作用;而且,miR-484可通过下调SORBS2并激活MEK/ERK通路参与乳腺癌的发生发展.
miR-484 Regulates the Proliferation,Metastasis and Autophagy of Breast Cancer Cells Through SORBS2/MEK-ERK Pathway
In order to analyze the expression of miR-484 in breast cancer tissues and cells,and its mecha-nism in breast cancer cell proliferation,metastasis and autophagy,the differentially expressed microRNAs(miRNAs)in breast cancer tissues were analyzed using GEO database,and the expression of miR-484 was detected by real-time fluorescence quantitative PCR in clinical breast cancer tissues and their paired para-cancerous tissues.The effects of miR-484 on the proliferation,metastasis and autophagy of MCF-7 breast cancer cells were detected by the miR-484 mimic and inhibitor.Then,the target gene of miR-484 was pre-dicted and verified,the Sorbin and SH3 domain-containing protein 2(SORBS2)overexpression vector was con-structed to detect the effects of SORBS2 on the proliferation,metastasis and autophagy of MCF-7 cells.Fi-nally,the protein contents of mitogen-activated extracellular signal-regulated kinase(MEK)/p-MEK and ex-tracellular signal-regulated kinase(ERK)/p-ERK in MCF-7 cells under the regulation of miR-484 were an-alyzed by Western-blot.The results showed that miR-484 was highly expressed in breast cancer tissues and cells,and improved the proliferation,migration,invasion of breast cancer cells and reduced their autophagy ability.Meanwhile,miR-484 could down-regulate SORBS2 and activate MEK/ERK pathway to participate in the occurrence and development of breast cancer.
breast cancermiR-484Sorbin and SH3 domain-containing protein 2(SORBS2)metastasisautophagy
万方数据
维普
