Gharakhyli Elaheh Asghari, Tabar Molla Hassan Agheel, Alipour Majid, Vahidi Sogand, Samadani Ali Akbar
Department of Microbiology, Gorgan Branch, Islamic Azad University, Gorgan, Iran.
Department of Cell and Molecular Biology, Babol Branch, Islamic Azad University, Babol, Iran.
Horm Mol Biol Clin Investig. 2023 Feb 28;44(3):271-276. doi: 10.1515/hmbci-2022-0045. eCollection 2023 Sep 1.
MicroRNA expression disruptions play an important function in the expansion of gastric cancer. Previous investigation has indicated that miR-372-5p doing as an oncogene in several malignancies. CDX1 and CDX2, as target genes of miR-372-5p, play the role of tumor suppressors and oncogenes in gastric cancer cells, respectively. The current investigation explored the effects of miR-372-5p regulation on CDX2 and CDX1 in AGS cell lines and studied their molecular mechanism.
hsa-miR-372-5p miRCURY LNA miRNA Inhibitors and Mimic were transfected into AGS cell line. The cell viability and cell cycle calculation were defined by MTT assay and flow cytometry, respectively. The Expression levels of miR-372-5p, CDX1, CDX2 and transfection efficiency were measured using Real-time PCR. Statistical investigation p values <0.05 were considered to be meaningful.
miR-372-5p particularly was upregulated in control cells and also after transfection by mimic. While its expression was reduced by the inhibitor. Upregulation of miR-372-5p remarkably increased cell growth and led to accumulation in the G2/M phase, although the inhibitor decreased cell growth and accumulation in the S phase. Accordingly, upregulation of miR-372-5p increased CDX2 and decreased CDX1 expression. By inhibition of miR-372-5p, expression of CDX2 was decreased and expression of CDX1 was increased.
Up and down-regulation of miR-372-5P has a potential effect on the expression levels of its target genes, CDX1 and CDX22. Accordingly, the downregulation of miR-372-5p may be assumed as a possible therapeutic target in treating gastric cancer.
微小RNA表达紊乱在胃癌进展中发挥重要作用。先前的研究表明,miR - 372 - 5p在多种恶性肿瘤中作为癌基因发挥作用。CDX1和CDX2作为miR - 372 - 5p的靶基因,在胃癌细胞中分别发挥肿瘤抑制因子和癌基因的作用。本研究探讨了miR - 372 - 5p调控对AGS细胞系中CDX2和CDX1的影响,并研究其分子机制。
将hsa - miR - 372 - 5p微小RNA抑制物和模拟物转染到AGS细胞系中。分别通过MTT法和流式细胞术测定细胞活力和细胞周期。使用实时定量PCR测量miR - 372 - 5p、CDX1、CDX2的表达水平和转染效率。统计学分析中,p值<0.05被认为具有统计学意义。
miR - 372 - 5p在对照细胞中以及转染模拟物后均显著上调。而其表达被抑制剂降低。miR - 372 - 5p的上调显著促进细胞生长,并导致G2/M期的积累,尽管抑制剂降低了细胞生长并导致S期的积累。相应地,miR - 372 - 5p的上调增加了CDX2的表达并降低了CDX1的表达。通过抑制miR - 372 - 5p,CDX2的表达降低,CDX1的表达增加。
miR - 372 - 5p的上调和下调对其靶基因CDX1和CDX2的表达水平具有潜在影响。因此,miR - 372 - 5p的下调可能被认为是治疗胃癌的一个潜在治疗靶点。