■迷迭香酸(RA),一种天然的酚酸,展现出有希望的抗癌特性。microRNA(miRNA)的异常表达调节基因表达,并在TNBC中作为致癌或肿瘤抑制因子发挥作用。然而,RA中miR-30a-5p对BCL2L11在MDA-MB-231诱导的乳腺癌干细胞样细胞(BCSCs)进展过程中的生物学作用及其调控机制尚未阐明。
■观察RA是否抑制miR-30a-5p对BCL2L11基因的沉默作用,促进BCSC细胞凋亡。
■我们评估了迁移,菌落形成,扩散,细胞周期,和使用伤口愈合测定法治疗RA后BCSCs的凋亡,集落形成试验,CCK-8测定,和流式细胞术,分别。BCL-2,Bax的mRNA和蛋白水平的表达,通过实时聚合酶链反应和Westernblot获得RA治疗后BCSCs中的BCL2L11和P53基因。通过高通量测序分析BCSC中的差异miRNA表达。利用Targetscan预测miR-30a-5p的靶标。双荧光素酶报告系统用于miR-30a-5p靶标的验证。
■伤口愈合试验,集落形成试验,CCK-8测定,细胞周期检测结果显示RA抑制细胞迁移,BCSC的集落形成和活力,细胞周期阻滞在G0-G1期。在最高剂量的RA,我们注意到细胞萎缩,而在100μg/mLRA时的阻滞率超过200μg/mLRA时的阻滞率。凋亡细胞早期出现(膜相关蛋白VFITC+,PI-)或晚期(膜相关蛋白VFITC+,PI+)在施用200μg/mLRA后,使用高通量测序来比较miRNA表达的差异,我们检测到miR-30a-5p表达下调,双荧光素酶报告基因分析结果表明BCL2L11是miR-30a-5p的直接靶标。
■RA抑制miR-30a-5p对BCL2L11基因的沉默作用,增强BCSC细胞凋亡。
UNASSIGNED: Rosmarinic acid (RA), a natural phenolic acid, exhibits promising anti-cancer properties. The abnormal expression of microRNA (miRNA) regulates the gene expression and plays a role as an oncogenic or tumor suppressor in TNBC. However, the biological role of RA in miR-30a-5p on BCL2L11 during MDA-MB-231 induced breast cancer stem-like cells (
BCSCs) progression and its regulatory mechanism have not been elucidated.
UNASSIGNED: To investigate whether RA inhibited the silencing effect of miR-30a-5p on the BCL2L11 gene and promoted apoptosis in
BCSCs.
UNASSIGNED: We assessed the migration, colony formation, proliferation, cell cycle, and apoptosis of BCSCs after RA treatment using the wound-healing assay, colony formation assay, CCK-8 assay, and flow cytometry, respectively. The expression of mRNA and protein levels of BCL-2, Bax, BCL2L11, and P53 genes in
BCSCs after RA treatment was obtained by real-time polymerase chain reaction and Western blot. Differential miRNA expression in
BCSCs was analyzed by high-throughput sequencing. Targetscan was utilized to predict the targets of miR-30a-5p. The dual luciferase reporter system was used for validation of the miR-30a-5p target.
UNASSIGNED: Wound-healing assay, colony formation assay, CCK-8 assay, and cell cycle assay results showed that RA inhibited migration, colony formation and viability of
BCSCs, and cell cycle arrest in the G0-G1 phase. At the highest dose of RA, we noticed cell atrophy, while the arrest rate at 100 μg/mL RA surpassed that at 200 μg/mL RA. Apoptotic cells appeared early (Membrane Associated Protein V FITC+, PI-) or late (Membrane Associated Protein V FITC+, PI+) upon administration of 200 μg/mL RA, Using high-throughput sequencing to compare the differences in miRNA expression, we detected downregulation of miR-30a-5p expression, and the results of dual luciferase reporter gene analysis indicated that BCL2L11 was a direct target of miR-30a-5p.
UNASSIGNED: RA inhibited the silencing effect of miR-30a-5p on the BCL2L11 gene and enhanced apoptosis in BCSCs.