Hosseinahli Nayer, Zeinali Tahereh, Hosseinahli Nasrin, Karimi Leila, Shanehbandi Dariush, Mansoori Behzad, Mohammadi Ali, Kazemi Tohid, Hajiasgharzadeh Khalil, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Gastrointestinal and Liver Diseases Research Center, Guilan University of Medical Sciences, Rasht, Iran.
Adv Pharm Bull. 2022 Jan;12(1):183-190. doi: 10.34172/apb.2022.020. Epub 2020 Oct 20.
Gastric cancer (GC) is one of the main causes of death from diseases, especially in developing countries. MicroRNAs (miRNAs) are important modulators of the messenger RNAs expression. Among these miRNAs, MiR-143 is a tumor suppressor miRNA and its irregular expression has been revealed in a diversity of malignancies such as GC. In this study, we have attempted to restore the miR-143 expression in MKN-45 cells by introducing pCMV-miR-143 plasmid vectors. The consequences of exogenous expression of miR-143 on cell proliferation and migration were assessed by MTT and scratch tests, respectively. In addition, the DAPI staining assay was applied for apoptosisquantification. Following miR-143 transfection, the changes in K-Ras, C-Myc, MMP9, Bax, Caspase-3, and Caspase-9 mRNA levels were assessed. The results indicated that the enhanced expression of miR-143 had negative effects on MKN-45 cells proliferation and invasion. Moreover, decreased expressions of K-Ras, MMP9, and C-Myc and up-regulation of Bax, Caspase-3, and Caspase-9 as downstream targets of miR-143 were recognized. These experimental results indicate that reversing the miR-143 expression, by novel techniques, including miRNA replacement could be considered as an efficient approach to reduce cell survival and metastasis.
胃癌(GC)是主要的致死疾病之一,在发展中国家尤为如此。微小RNA(miRNA)是信使RNA表达的重要调节因子。在这些miRNA中,MiR-143是一种肿瘤抑制性miRNA,其异常表达已在多种恶性肿瘤如胃癌中被发现。在本研究中,我们试图通过导入pCMV-miR-143质粒载体来恢复MKN-45细胞中MiR-143的表达。分别通过MTT和划痕试验评估了MiR-143外源表达对细胞增殖和迁移的影响。此外,采用DAPI染色法进行凋亡定量分析。在转染MiR-143后,评估了K-Ras、C-Myc、MMP9、Bax、Caspase-3和Caspase-9 mRNA水平的变化。结果表明,MiR-143表达增强对MKN-45细胞的增殖和侵袭具有负面影响。此外,还发现K-Ras、MMP9和C-Myc的表达降低,以及作为MiR-143下游靶点的Bax、Caspase-3和Caspase-9的表达上调。这些实验结果表明,通过包括miRNA替代在内的新技术逆转MiR-143的表达,可被视为一种降低细胞存活和转移的有效方法。