miR-484通过SORBS2/MEK-ERK通路调控乳腺癌细胞增殖、转移和自噬

miR-484 Regulates the Proliferation, Metastasis and Autophagy of Breast Cancer Cells Through SORBS2/MEK-ERK Pathway

摘要

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。

Abstract

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.

关键词

乳腺癌 / miR-484 / Sorbin和SH3结构域包含蛋白2 (SORBS2) / 转移 / 自噬

Key words

breast cancer / miR-484 / Sorbin and SH3 domain-containing protein 2 (SORBS2) / metastasis / autophagy