The Relationship between Autophagy Process and Expression of MicroRNA-146a-5p in MKN-45 and MCF-7 Cell Lines
-
Atefe Alirezaee
,
Ahmad Zavaran Hosseini
,
Sara Soudi
,
Nazanin Atieh Kazemi-Sefat
,
Mohammad Mahdi Jafari
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.
1. Kim YS, Shin JH, Bae MK, Lee CY, Kim DJ, Chung KY, et al. Autophagy activity in pulmonary metastatic tumor tissues from colorectal cancer: a pilot study. Yonsei Medical Journal. 2014;55(6):1484-8.
2. Tsuchihara K, Fujii S, Esumi H. Autophagy and cancer: dynamism of the metabolism of tumor cells and tissues. Cancer letters. 2009;278(2):130-8.
3. Jeda AS, Ghabeshi S, Jazaeri EO, Araiinejad M, Sheikholeslami F, Abdoli M, et al. Autophagy Modulation and Cancer Combination Therapy: A Smart Approach in Cancer Therapy. Cancer Treatment and Research Communications. 2022:100512.
4. Borhani S. Autophagy in Cancer Therapy: Advances and Prospects. Current Molecular Biology Reports. 2023;9(1):1-7.
5. Mehrpour M, Esclatine A, Beau I, Codogno P. Overview of macroautophagy regulation in mammalian cells. Cell research. 2010;20(7):748-62.
6. Zhou Y, Manghwar H, Hu W, Liu F. Degradation Mechanism of Autophagy-Related Proteins and Research Progress. International journal of molecular sciences. 2022;23(13):7301.
7. Bartel DP. MicroRNAs: target recognition and regulatory functions. cell. 2009;136(2):215-33.
8. Gozuacik D, Akkoc Y, Ozturk DG, Kocak M. Autophagy-regulating microRNAs and cancer. Frontiers in oncology. 2017;7:65.
9. Zhang H, Liang J, Chen N. The Potential Role of miRNA-Regulated Autophagy in Alzheimer’s Disease. International journal of molecular sciences. 2022;23(14):7789.
10. Wei J, Wang X, Yu D, Tu Y, Yu Y. MicroRNA-mediated autophagy and drug resistance in cancer: mechanisms and therapeutic strategies. Discover Oncology. 2024;15(1):662.
11. Lujambio A, Lowe SW. The microcosmos of cancer. Nature. 2012;482(7385):347-55.
12. Chen S, Wang Y, Li D, Wang H, Zhao X, Yang J, et al. Mechanisms controlling microRNA expression in tumor.
Cells. 2022;11(18):2852.
13. Chakrabortty A, Patton DJ, Smith BF, Agarwal P. miRNAs: potential as biomarkers and therapeutic targets for cancer. Genes. 2023;14(7):1375.
14. Dimopoulos K, Gimsing P, Grønbæk K. Aberrant micro RNA expression in multiple myeloma. European journal of haematology. 2013;91(2):95-105.
15. Li L, Chen XP, Li YJ. MicroRNA‐146a and human disease. Scandinavian journal of immunology. 2010;71(4):227-31.
16. Hou Z, Xie L, Yu L, Qian X, Liu B. MicroRNA-146a is down-regulated in gastric cancer and regulates cell proliferation and apoptosis. Medical oncology. 2012;29(2):886-92.
17. Si C, Yu Q, Yao Y. Effect of miR-146a-5p on proliferation and metastasis of triple-negative breast cancer via regulation of SOX5. Experimental and Therapeutic Medicine. 2018;15(5):4515-21.
18. Xiao B, Zhu E-D, Li N, Lu D-S, Li W, Li B-S, et al. Increased miR-146a in gastric cancer directly targets SMAD4 and is involved in modulating cell proliferation and apoptosis. Oncology reports. 2012;27(2):559-66.
19. Karimi M, Mohammadnia A, Amini MA, Shamekh AG, Derakhshanfar E, Hosseini F. Overexpression of miR-146a and miR-155 are Potentially Biomarkers and Predict Unfavorable Relationship between Gastric Cancer and Helicobacter pylori Infection. Chonnam Medical Journal. 2023;59(3):167.
20. Iacona JR, Lutz CS. miR‐146a‐5p: expression, regulation, and functions in cancer. Wiley Interdisciplinary Reviews: RNA. 2019;10(4):e1533.
21. Shi L, Xu Z, Wu G, Chen X, Huang Y, Wang Y, et al. Up-regulation of miR-146a increases the sensitivity of non-small cell lung cancer to DDP by downregulating cyclin J. BMC cancer. 2017;17(1):1-14.
22. Pang L, Lu J, Huang J, Xu C, Li H, Yuan G, et al. Upregulation of miR‑146a increases cisplatin sensitivity of the non‑small cell lung cancer A549 cell line by targeting JNK‑2. Oncology Letters. 2017;14(6):7745-52.
23. Luo Z, Li X, Zhao Z, Yang X, Xiao S, Zhou Y. MicroRNA-146a affects the chemotherapeutic sensitivity and prognosis of advanced gastric cancer through the regulation of LIN52. Oncology Letters. 2017;13(3):1386-92.
24. Pu J, Wu S, Xie H, Li Y, Yang Z, Wu X, et al. miR-146a Inhibits dengue-virus-induced autophagy by targeting TRAF6. Archives of virology. 2017;162(12):3645-59.
25. Zhu W, Qu H, Xu K, Jia B, Li H, Du Y, et al. Differences in the starvation-induced autophagy response in MDA-MB-231 and MCF-7 breast cancer cells. Animal Cells and Systems. 2017;21(3):190-8.
26. Sa-Nongdej W, Chongthammakun S, Songthaveesin C. Nutrient starvation induces apoptosis and autophagy in C6 glioma stem-like cells. Heliyon. 2021;7(2).
27. Marinković M, Šprung M, Buljubašić M, Novak I. Autophagy modulation in cancer: current knowledge on action and therapy. Oxidative Medicine and Cellular Longevity. 2018;2018.
28. Alva A, Gultekin S, Baehrecke E. Autophagy in human tumors: cell survival or death? Cell Death & Differentiation. 2004;11(9):1046-8.
29. Macintosh RL, Ryan KM, editors. Autophagy in tumour cell death. Seminars in Cancer Biology; 2013: Elsevier.
30. Bergmann A. Autophagy and cell death: no longer at odds. Cell. 2007;131(6):1032-4.
31. Jiang P, Mizushima N. Autophagy and human diseases. Cell research. 2014;24(1):69-79.
32. Hassan AMIA, Zhao Y, Chen X, He C. Blockage of autophagy for cancer therapy: a comprehensive review. International journal of molecular sciences. 2024;25(13):7459.
33. Park H, Huang X, Lu C, Cairo MS, Zhou X. MicroRNA-146a and microRNA-146b regulate human dendritic cell apoptosis and cytokine production by targeting TRAF6 and IRAK1 proteins. Journal of Biological Chemistry. 2015;290(5):2831-41.
34. Gao W, Hua J, Jia Z, Ding J, Han Z, Dong Y, et al. Expression of miR‑146a‑5p in breast cancer and its role in proliferation of breast cancer cells. Oncology letters. 2018;15(6):9884-8.
35. Yang X, Liu R. Long non-coding RNA HCG18 promotes gastric cancer progression by regulating miRNA-146a-5p/tumor necrosis factor receptor-associated factor 6 axis. Bioengineered. 2022;13(3):6781-93.
36. Kuma A, Hatano M, Matsui M, Yamamoto A, Nakaya H, Yoshimori T, et al. The role of autophagy during the early neonatal starvation period. Nature. 2004;432(7020):1032-6.
37. Rikiishi H. Novel insights into the interplay between apoptosis and autophagy. International journal of cell biology. 2012;2012(1):317645.
38. Li M, Gao P, Zhang J. Crosstalk between autophagy and apoptosis: potential and emerging therapeutic targets for cardiac diseases. International journal of molecular sciences. 2016;17(3):332.
39. Hu Z-q, Li Q, Hu Z-h, Liu H-c, Rao C-l, Zhang M-j, et al. MicroRNA-146a inhibits autophagy to maintain the intracellular survival of Burkholderia pseudomallei by targeting LIPA. Microbial Pathogenesis. 2021;158:104969.
40. Shang Y, Liu Q, Wang L, Qiu X, Chen Y, An J. microRNA-146a-5p negatively modulates PM2. 5 caused inflammation in THP-1 cells via autophagy process. Environmental Pollution. 2021;268:115961.
41. Li M-w, Gao L, Dang Y-w, Li P, Li Z-y, Chen G, et al. Protective potential of miR-146a-5p and its underlying molecular mechanism in diverse cancers: a comprehensive meta-analysis and bioinformatics analysis. Cancer Cell International. 2019;19:1-21.
42. Noorolyai S, Baghbani E, Shanehbandi D, Shahgoli VK, Kojabad AB, Mansoori B, et al. miR-146a-5p and miR-193a-5p Synergistically Inhibited the Proliferation of Human Colorectal Cancer Cells (HT-29 cell line) through ERK Signaling Pathway. Advanced Pharmaceutical Bulletin. 2021;11(4):755.
43. Luo J, Si Z-Z, Li T, Li J-Q, Zhang Z-Q, Chen G-S, et al. MicroRNA-146a-5p enhances radiosensitivity in hepatocellular carcinoma through replication protein A3-induced activation of the DNA repair pathway. American Journal of Physiology-Cell Physiology. 2019;316(3):C299-C311.
44. Yuwen D, Sheng B, Liu J, Wenyu W, Shu Y. MiR-146a-5p level in serum exosomes predicts therapeutic effect of cisplatin in non-small cell lung cancer. Eur Rev Med Pharmacol Sci. 2017;21(11):2650-8.
45. Chen Z, Gu Q, Chen R. miR-146a-5p regulates autophagy and NLRP3 inflammasome activation in epithelial barrier damage in the in vitro cell model of ulcerative colitis through the RNF8/Notch1/mTORC1 pathway. Immunobiology. 2023;228(4):152386.
46. Zhao H, Li Q, Pang J, Jin H, Li H, Yang X. Blocking autophagy enhances the pro-apoptotic effect of bufalin on human gastric cancer cells through endoplasmic reticulum stress. Biology Open. 2017;6(10):1416-22.
2. Tsuchihara K, Fujii S, Esumi H. Autophagy and cancer: dynamism of the metabolism of tumor cells and tissues. Cancer letters. 2009;278(2):130-8.
3. Jeda AS, Ghabeshi S, Jazaeri EO, Araiinejad M, Sheikholeslami F, Abdoli M, et al. Autophagy Modulation and Cancer Combination Therapy: A Smart Approach in Cancer Therapy. Cancer Treatment and Research Communications. 2022:100512.
4. Borhani S. Autophagy in Cancer Therapy: Advances and Prospects. Current Molecular Biology Reports. 2023;9(1):1-7.
5. Mehrpour M, Esclatine A, Beau I, Codogno P. Overview of macroautophagy regulation in mammalian cells. Cell research. 2010;20(7):748-62.
6. Zhou Y, Manghwar H, Hu W, Liu F. Degradation Mechanism of Autophagy-Related Proteins and Research Progress. International journal of molecular sciences. 2022;23(13):7301.
7. Bartel DP. MicroRNAs: target recognition and regulatory functions. cell. 2009;136(2):215-33.
8. Gozuacik D, Akkoc Y, Ozturk DG, Kocak M. Autophagy-regulating microRNAs and cancer. Frontiers in oncology. 2017;7:65.
9. Zhang H, Liang J, Chen N. The Potential Role of miRNA-Regulated Autophagy in Alzheimer’s Disease. International journal of molecular sciences. 2022;23(14):7789.
10. Wei J, Wang X, Yu D, Tu Y, Yu Y. MicroRNA-mediated autophagy and drug resistance in cancer: mechanisms and therapeutic strategies. Discover Oncology. 2024;15(1):662.
11. Lujambio A, Lowe SW. The microcosmos of cancer. Nature. 2012;482(7385):347-55.
12. Chen S, Wang Y, Li D, Wang H, Zhao X, Yang J, et al. Mechanisms controlling microRNA expression in tumor.
Cells. 2022;11(18):2852.
13. Chakrabortty A, Patton DJ, Smith BF, Agarwal P. miRNAs: potential as biomarkers and therapeutic targets for cancer. Genes. 2023;14(7):1375.
14. Dimopoulos K, Gimsing P, Grønbæk K. Aberrant micro RNA expression in multiple myeloma. European journal of haematology. 2013;91(2):95-105.
15. Li L, Chen XP, Li YJ. MicroRNA‐146a and human disease. Scandinavian journal of immunology. 2010;71(4):227-31.
16. Hou Z, Xie L, Yu L, Qian X, Liu B. MicroRNA-146a is down-regulated in gastric cancer and regulates cell proliferation and apoptosis. Medical oncology. 2012;29(2):886-92.
17. Si C, Yu Q, Yao Y. Effect of miR-146a-5p on proliferation and metastasis of triple-negative breast cancer via regulation of SOX5. Experimental and Therapeutic Medicine. 2018;15(5):4515-21.
18. Xiao B, Zhu E-D, Li N, Lu D-S, Li W, Li B-S, et al. Increased miR-146a in gastric cancer directly targets SMAD4 and is involved in modulating cell proliferation and apoptosis. Oncology reports. 2012;27(2):559-66.
19. Karimi M, Mohammadnia A, Amini MA, Shamekh AG, Derakhshanfar E, Hosseini F. Overexpression of miR-146a and miR-155 are Potentially Biomarkers and Predict Unfavorable Relationship between Gastric Cancer and Helicobacter pylori Infection. Chonnam Medical Journal. 2023;59(3):167.
20. Iacona JR, Lutz CS. miR‐146a‐5p: expression, regulation, and functions in cancer. Wiley Interdisciplinary Reviews: RNA. 2019;10(4):e1533.
21. Shi L, Xu Z, Wu G, Chen X, Huang Y, Wang Y, et al. Up-regulation of miR-146a increases the sensitivity of non-small cell lung cancer to DDP by downregulating cyclin J. BMC cancer. 2017;17(1):1-14.
22. Pang L, Lu J, Huang J, Xu C, Li H, Yuan G, et al. Upregulation of miR‑146a increases cisplatin sensitivity of the non‑small cell lung cancer A549 cell line by targeting JNK‑2. Oncology Letters. 2017;14(6):7745-52.
23. Luo Z, Li X, Zhao Z, Yang X, Xiao S, Zhou Y. MicroRNA-146a affects the chemotherapeutic sensitivity and prognosis of advanced gastric cancer through the regulation of LIN52. Oncology Letters. 2017;13(3):1386-92.
24. Pu J, Wu S, Xie H, Li Y, Yang Z, Wu X, et al. miR-146a Inhibits dengue-virus-induced autophagy by targeting TRAF6. Archives of virology. 2017;162(12):3645-59.
25. Zhu W, Qu H, Xu K, Jia B, Li H, Du Y, et al. Differences in the starvation-induced autophagy response in MDA-MB-231 and MCF-7 breast cancer cells. Animal Cells and Systems. 2017;21(3):190-8.
26. Sa-Nongdej W, Chongthammakun S, Songthaveesin C. Nutrient starvation induces apoptosis and autophagy in C6 glioma stem-like cells. Heliyon. 2021;7(2).
27. Marinković M, Šprung M, Buljubašić M, Novak I. Autophagy modulation in cancer: current knowledge on action and therapy. Oxidative Medicine and Cellular Longevity. 2018;2018.
28. Alva A, Gultekin S, Baehrecke E. Autophagy in human tumors: cell survival or death? Cell Death & Differentiation. 2004;11(9):1046-8.
29. Macintosh RL, Ryan KM, editors. Autophagy in tumour cell death. Seminars in Cancer Biology; 2013: Elsevier.
30. Bergmann A. Autophagy and cell death: no longer at odds. Cell. 2007;131(6):1032-4.
31. Jiang P, Mizushima N. Autophagy and human diseases. Cell research. 2014;24(1):69-79.
32. Hassan AMIA, Zhao Y, Chen X, He C. Blockage of autophagy for cancer therapy: a comprehensive review. International journal of molecular sciences. 2024;25(13):7459.
33. Park H, Huang X, Lu C, Cairo MS, Zhou X. MicroRNA-146a and microRNA-146b regulate human dendritic cell apoptosis and cytokine production by targeting TRAF6 and IRAK1 proteins. Journal of Biological Chemistry. 2015;290(5):2831-41.
34. Gao W, Hua J, Jia Z, Ding J, Han Z, Dong Y, et al. Expression of miR‑146a‑5p in breast cancer and its role in proliferation of breast cancer cells. Oncology letters. 2018;15(6):9884-8.
35. Yang X, Liu R. Long non-coding RNA HCG18 promotes gastric cancer progression by regulating miRNA-146a-5p/tumor necrosis factor receptor-associated factor 6 axis. Bioengineered. 2022;13(3):6781-93.
36. Kuma A, Hatano M, Matsui M, Yamamoto A, Nakaya H, Yoshimori T, et al. The role of autophagy during the early neonatal starvation period. Nature. 2004;432(7020):1032-6.
37. Rikiishi H. Novel insights into the interplay between apoptosis and autophagy. International journal of cell biology. 2012;2012(1):317645.
38. Li M, Gao P, Zhang J. Crosstalk between autophagy and apoptosis: potential and emerging therapeutic targets for cardiac diseases. International journal of molecular sciences. 2016;17(3):332.
39. Hu Z-q, Li Q, Hu Z-h, Liu H-c, Rao C-l, Zhang M-j, et al. MicroRNA-146a inhibits autophagy to maintain the intracellular survival of Burkholderia pseudomallei by targeting LIPA. Microbial Pathogenesis. 2021;158:104969.
40. Shang Y, Liu Q, Wang L, Qiu X, Chen Y, An J. microRNA-146a-5p negatively modulates PM2. 5 caused inflammation in THP-1 cells via autophagy process. Environmental Pollution. 2021;268:115961.
41. Li M-w, Gao L, Dang Y-w, Li P, Li Z-y, Chen G, et al. Protective potential of miR-146a-5p and its underlying molecular mechanism in diverse cancers: a comprehensive meta-analysis and bioinformatics analysis. Cancer Cell International. 2019;19:1-21.
42. Noorolyai S, Baghbani E, Shanehbandi D, Shahgoli VK, Kojabad AB, Mansoori B, et al. miR-146a-5p and miR-193a-5p Synergistically Inhibited the Proliferation of Human Colorectal Cancer Cells (HT-29 cell line) through ERK Signaling Pathway. Advanced Pharmaceutical Bulletin. 2021;11(4):755.
43. Luo J, Si Z-Z, Li T, Li J-Q, Zhang Z-Q, Chen G-S, et al. MicroRNA-146a-5p enhances radiosensitivity in hepatocellular carcinoma through replication protein A3-induced activation of the DNA repair pathway. American Journal of Physiology-Cell Physiology. 2019;316(3):C299-C311.
44. Yuwen D, Sheng B, Liu J, Wenyu W, Shu Y. MiR-146a-5p level in serum exosomes predicts therapeutic effect of cisplatin in non-small cell lung cancer. Eur Rev Med Pharmacol Sci. 2017;21(11):2650-8.
45. Chen Z, Gu Q, Chen R. miR-146a-5p regulates autophagy and NLRP3 inflammasome activation in epithelial barrier damage in the in vitro cell model of ulcerative colitis through the RNF8/Notch1/mTORC1 pathway. Immunobiology. 2023;228(4):152386.
46. Zhao H, Li Q, Pang J, Jin H, Li H, Yang X. Blocking autophagy enhances the pro-apoptotic effect of bufalin on human gastric cancer cells through endoplasmic reticulum stress. Biology Open. 2017;6(10):1416-22.
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| Issue | Articles in Press | |
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| Keywords | ||
| Autophagy Autophagy-related Genes Breast cancer Gastric cancer MicroRNA MCF-7 cell line MKN-45 cell line miR-146a-5p | ||
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How to Cite
1.
Alirezaee A, Zavaran Hosseini A, Soudi S, Kazemi-Sefat NA, Jafari MM. The Relationship between Autophagy Process and Expression of MicroRNA-146a-5p in MKN-45 and MCF-7 Cell Lines. Iran J Allergy Asthma Immunol. 2025;:1-10.
