Nie Fangfang, Zhang Qinfang, Ma WeiNa, Yan Jun
Department of Oncology, Jiading District Central Hospital Affiliated Shanghai University of Medicine & Health Sciences, No. 1 Chengbei Road, Jiading District, Shanghai, 201800, China.
Department of Pharmacy, Jiading District Central Hospital Affiliated Shanghai University of Medicine & Health Sciences, Shanghai, 201800, China.
Discov Oncol. 2024 May 8;15(1):146. doi: 10.1007/s12672-024-01004-1.
Epithelial-mesenchymal transition (EMT) plays an important role in malignant progression of Triple-negative breast cancer (TNBC). Many studies have confirmed that miRNA-200c-3p is related to EMT. And we found that it is involved in the regulation of EMT, but the exact mechanism is unclear. CRKL is highly expressed in a variety of tumors and plays a role in EMT. In this study, the potential targets of miRNA-200c-3p were searched in miRPathDB, Targetscan and PicTar. And there are 68 potential targets at the intersection of the three databases. Then, bioinformatics and text mining performed by Coremine Medica, and found that among 68 potential targets, CRKL has the strongest correlation with EMT in TNBC. Therefore, we speculated that miRNA-200c-3p involvement in EMT might be related to CRKL. To verify miRNA-200c-3p inhibits the malignant phenotype of TNBC by regulating CRKL, RT‒PCR, western blotting, Clonal formation assays,CCK-8 proliferation assays, transwell invasion assays, Luciferase reporter assay and nude mouse transplantation tumor assay were performed. In this study, we found that miRNA-200c-3p is under-expressed and EMT-related genes are up-regulated in TNBC, and miRNA-200c-3p can inhibit cancer cell proliferation, invasion and the expression of EMT-related genes and proteins in TNBC. Further research confirmed that miRNA-200c-3p could inhibit EMT by inhibiting the expression of CRKL that directly combining CRKL gene.
上皮-间质转化（EMT）在三阴性乳腺癌（TNBC）的恶性进展中起重要作用。许多研究证实miRNA-200c-3p与EMT相关。并且我们发现它参与EMT的调控，但确切机制尚不清楚。CRKL在多种肿瘤中高表达并在EMT中发挥作用。在本研究中，在miRPathDB、Targetscan和PicTar中搜索miRNA-200c-3p的潜在靶标。三个数据库的交集处有68个潜在靶标。然后，通过Coremine Medica进行生物信息学和文本挖掘，发现在68个潜在靶标中，CRKL与TNBC中EMT的相关性最强。因此，我们推测miRNA-200c-3p参与EMT可能与CRKL有关。为了验证miRNA-200c-3p通过调节CRKL抑制TNBC的恶性表型，进行了RT-PCR、蛋白质免疫印迹、克隆形成试验、CCK-8增殖试验、Transwell侵袭试验、荧光素酶报告基因试验和裸鼠移植瘤试验。在本研究中，我们发现miRNA-200c-3p在TNBC中表达下调且EMT相关基因上调，并且miRNA-200c-3p可以抑制TNBC中癌细胞的增殖、侵袭以及EMT相关基因和蛋白质的表达。进一步研究证实miRNA-200c-3p可通过抑制直接结合CRKL基因的CRKL表达来抑制EMT。
