Inhibition of miR-20a-5p Suppresses Epithelial-Mesenchymal Transition of Colorectal Cancer Cells Through GJA1.

IF 2.4 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Biotechnology Pub Date : 2024-11-22 DOI:10.1007/s12033-024-01315-2
Lu Zhang, Jun-Bin Wang, Zhen-Yuan Gao, Xiao Wu, Hai-Rong Zhou
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Abstract

This study was designed to clarify the role of GJA1 in colorectal cancer. qPCR was adopted to detect the GJA1 and miR-20a-5p expression levels in tumor tissues and cells; and EdU, Transwell assay, Scratch test to examine the migration, invasion, and proliferation of colorectal cancer cells. The EMT-related protein expression was measured by immunofluorescence and western Blot. The binding relationship between GJA1 and miR-20a-5p was examined using dual luciferase reporting subsystem. In situ hybridization was utilized to examine the miR-20a-5p expression in tumor tissues and metastases. Rescue experiments were performed by simultaneous transfection of sh-GJA1 inhibitor and miR-20a-5p inhibitor. The miR-20a-5p expression was high and the GJA1 expression was low in colorectal cancer tissues and cells. A targeting relationship was found in GJA1 and miR-20a-5p targets. The invasion, migration, and proliferation of colorectal cancer cells can be inhibited by overexpression of GJA1. Meanwhile, overexpression of GJA1 markedly elevated the e-cadherin expression, but reduced the levels of vimentin, α-SMA and n-cadherin expression. miR-20a-5p inhibitor + sh-GJA1 promoted the invasion, migration, and proliferation of colon cancer cells and EMT process. Overall, miR-20a-5p could target GJA1 to down-regulate the GJA1 expression, thereby regulating the EMT response, and ultimately promoting the progression of colorectal cancer.

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抑制 miR-20a-5p 可通过 GJA1 抑制结直肠癌细胞的上皮-间质转化
本研究旨在阐明 GJA1 在结直肠癌中的作用。研究采用 qPCR 方法检测 GJA1 和 miR-20a-5p 在肿瘤组织和细胞中的表达水平;采用 EdU、Transwell 试验和 Scratch 试验检测结直肠癌细胞的迁移、侵袭和增殖。免疫荧光和 Western Blot 检测了 EMT 相关蛋白的表达。利用双荧光素酶报告子系统检测了 GJA1 和 miR-20a-5p 之间的结合关系。利用原位杂交技术检测了肿瘤组织和转移灶中 miR-20a-5p 的表达。通过同时转染 sh-GJA1 抑制剂和 miR-20a-5p 抑制剂进行拯救实验。结果表明,在结直肠癌组织和细胞中,miR-20a-5p 的表达量较高,而 GJA1 的表达量较低。研究发现 GJA1 和 miR-20a-5p 的靶点存在靶向关系。过表达 GJA1 可抑制结直肠癌细胞的侵袭、迁移和增殖。miR-20a-5p抑制剂+sh-GJA1促进了结肠癌细胞的侵袭、迁移和增殖以及EMT过程。总之,miR-20a-5p可以靶向GJA1,下调GJA1的表达,从而调节EMT反应,最终促进结直肠癌的进展。
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来源期刊
Molecular Biotechnology
Molecular Biotechnology 医学-生化与分子生物学
CiteScore
4.10
自引率
3.80%
发文量
165
审稿时长
6 months
期刊介绍: Molecular Biotechnology publishes original research papers on the application of molecular biology to both basic and applied research in the field of biotechnology. Particular areas of interest include the following: stability and expression of cloned gene products, cell transformation, gene cloning systems and the production of recombinant proteins, protein purification and analysis, transgenic species, developmental biology, mutation analysis, the applications of DNA fingerprinting, RNA interference, and PCR technology, microarray technology, proteomics, mass spectrometry, bioinformatics, plant molecular biology, microbial genetics, gene probes and the diagnosis of disease, pharmaceutical and health care products, therapeutic agents, vaccines, gene targeting, gene therapy, stem cell technology and tissue engineering, antisense technology, protein engineering and enzyme technology, monoclonal antibodies, glycobiology and glycomics, and agricultural biotechnology.
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