The Relationship between Autophagy Process and Expression of MicroRNA-146a-5p in MKN-45 and MCF-7 Cell Lines

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 a...

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Published inIranian journal of allergy, asthma, and immunology Vol. 24; no. 4; p. 472
Main Authors Alirezaee, Atefe, Hosseini, Ahmad Zavaran, Soudi, Sara, Kazemi-Sefat, Nazanin Atieh, Jafari, Mohammad Mahdi
Format Journal Article
LanguageEnglish
Published Iran Tehran University of Medical Sciences 26.06.2025
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ISSN1735-1502
1735-5249
DOI10.18502/ijaai.v24i4.19128

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Abstract 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.  
AbstractList 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.
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.  
Author Soudi, Sara
Kazemi-Sefat, Nazanin Atieh
Hosseini, Ahmad Zavaran
Alirezaee, Atefe
Jafari, Mohammad Mahdi
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Issue 4
Keywords Autophagy-related Genes
miR-146a-5p
MicroRNA
MCF-7 cell line
Breast cancer
Autophagy
Gastric cancer
MKN-45 cell line
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StartPage 472
SubjectTerms Antineoplastic drugs
Apoptosis
Apoptosis - genetics
Autophagy
Autophagy - genetics
Breast cancer
Breast Neoplasms - genetics
Breast Neoplasms - pathology
Cancer therapies
Cell Line, Tumor
Chemotherapy
Female
Flow cytometry
Fluorescence microscopy
Gastric cancer
Gene Expression Regulation, Neoplastic
Humans
MCF-7 Cells
MicroRNAs
MicroRNAs - genetics
miRNA
Polymerase chain reaction
Radiation therapy
Stomach Neoplasms - genetics
Stomach Neoplasms - metabolism
Stomach Neoplasms - pathology
TRAF6 protein
Tumor cell lines
Tumor cells
Tumor necrosis factor receptors
Vacuoles
Western blotting
Title The Relationship between Autophagy Process and Expression of MicroRNA-146a-5p in MKN-45 and MCF-7 Cell Lines
URI https://www.ncbi.nlm.nih.gov/pubmed/40696733
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Volume 24
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