Alirezaee Atefe, Zavaran Hosseini Ahmad, Soudi Sara, Kazemi-Sefat Nazanin Atieh, Jafari Mohammad Mahdi
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Iran J Allergy Asthma Immunol. 2025 Jun 26;24(4):472-480.
After chemotherapy or radiation therapy, autophagy activity increases in tumor cells for the adaptation of the tumor cells to stress. Thus, disturbance in autophagy can enhance the effectiveness of anticancer drugs. On the other hand, recent findings highlight the importance of microRNAs (miRs) in autophagy, including miR-146a-5p. In gastric and breast cancer miR-146a-5p is frequently reduced, and more precise identification of its function in these cancers is needed. The aim of this study was to evaluate the relationship between miR-146a-5p and autophagy in MKN-45 (human stomach cancer cell line) and MCF-7(breast cancer cell line). The expression of miR-146a-5p in MKN-45 and MCF-7 cell lines was measured before and after induction of autophagy using real-time polymerase chain reaction (PCR). A flow cytometry assay was used for the apoptosis assay, and autophagy induction was approved. Also, the formation of autophagic vacuoles was ensured in cells by western blotting and fluorescence microscopy. Real-time PCR showed that miR-146a-5p level in starvation groups, during autophagy, was significantly lower than in control groups, and also tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) level, a key target of miR-146a-5p, in starvation groups, during autophagy, was more than control groups but it was significant only in the MCF-7 group. According to previous studies and the results of the present study, miR-146a-5p may be considered a negative regulator of autophagy. However, to confirm this, further studies are needed on different cancer cell lines.
化疗或放疗后,肿瘤细胞中的自噬活性会增加,以帮助肿瘤细胞适应应激状态。因此,自噬紊乱可增强抗癌药物的疗效。另一方面,最近的研究结果突出了微小RNA(miR)在自噬中的重要性,包括miR-146a-5p。在胃癌和乳腺癌中,miR-146a-5p常常表达降低,因此需要更精确地确定其在这些癌症中的功能。本研究的目的是评估miR-146a-5p与MKN-45(人胃癌细胞系)和MCF-7(乳腺癌细胞系)中自噬的关系。使用实时聚合酶链反应(PCR)在诱导自噬前后测量MKN-45和MCF-7细胞系中miR-146a-5p的表达。采用流式细胞术检测细胞凋亡情况,并证实自噬被诱导。此外,通过蛋白质印迹法和荧光显微镜检查确保细胞中自噬泡的形成。实时PCR结果显示,在自噬过程中,饥饿组的miR-146a-5p水平显著低于对照组,而miR-146a-5p的关键靶标肿瘤坏死因子受体(TNFR)相关因子6(TRAF6)在饥饿组自噬过程中的水平高于对照组,但仅在MCF-7组有显著差异。根据以往研究及本研究结果,miR-146a-5p可能被视为自噬的负调节因子。然而要证实这一点,则需要对不同癌细胞系开展进一步研究。