Pei Bei, Li Tongyang, Qian Qi, Fan Wenqiang, He Xiao, Zhu Yulan, Xu Lingyun
Changzhou No. 2 People's Hospital, Xinglong Lance, Changzhou, China.
Department of Breast Surgery, Changzhou No. 2 People's Hospital, Changzhou 213003, China.
Gland Surg. 2020 Jun;9(3):747-758. doi: 10.21037/gs-20-472.
Breast cancer metastasis is the main problem that affects the therapy and prognosis of breast cancer patients. Studies have indicated the role of microRNAs in breast cancer regulation, but the mechanisms are largely unknown.
In this study, we determined the expression of microRNA-30c-5p (miR-30c-5p) and coactosin-like protein 1 () gene in breast cancer tissues, and revealed their effects on breast cancer metastasis regulation. Breast cancer and paracancerous tissues were collected. Reverse transcriptase polymerase chain reaction RT-PCR) was used to analyze the expression of miR-30c-5p and , and breast cancer cell line (MCF-7) was employed to verify the relationship between miR-30c-5p and COTL1. Western blot analysis and immunofluorescence were used for proteins analysis and microfilament observation, respectively. A dual-luciferase reporter gene was used for microRNA-gene interaction assay.
The results showed that the expression of miR-30c-5p decreased, while the expression of COTL1 increased in breast cancer tissues. The results of luciferase reporting gene assay showed that, COTL1 was the target of miR-30c-5p. After miR-30c-5p was upregulated, the expression of COTL1 was reduced, microfilament arrangement was in disorder, and cell migration ability was inhibited. After miR-30c-5p was downregulated, the expression of COTL1 was increased, and the cell migration ability was enhanced. COTL1 protein expression levels were significantly higher in cancer tissues with lymph node metastasis.
These findings indicate that miR-30c-5p/COTL1 pathway regulates breast cancer metastasis and can be used as a potential therapy target.
乳腺癌转移是影响乳腺癌患者治疗和预后的主要问题。研究表明微小RNA在乳腺癌调控中发挥作用,但其机制尚不清楚。
在本研究中,我们检测了微小RNA-30c-5p(miR-30c-5p)和类肌动蛋白结合蛋白1(COTL1)基因在乳腺癌组织中的表达,并揭示了它们对乳腺癌转移调控的影响。收集乳腺癌组织及癌旁组织。采用逆转录聚合酶链反应(RT-PCR)分析miR-30c-5p和COTL1的表达,并利用乳腺癌细胞系(MCF-7)验证miR-30c-5p与COTL1之间的关系。分别采用蛋白质免疫印迹分析和免疫荧光进行蛋白质分析和微丝观察。采用双荧光素酶报告基因进行微小RNA-基因相互作用检测。
结果显示,乳腺癌组织中miR-30c-5p表达降低,而COTL1表达升高。荧光素酶报告基因检测结果表明,COTL1是miR-30c-5p的靶标。上调miR-30c-5p后,COTL1表达降低,微丝排列紊乱,细胞迁移能力受到抑制。下调miR-30c-5p后,COTL1表达增加,细胞迁移能力增强。有淋巴结转移的癌组织中COTL1蛋白表达水平显著更高。
这些研究结果表明,miR-30c-5p/COTL1通路调控乳腺癌转移,可作为潜在的治疗靶点。