The Role of Hyperthermia in Enhancing Anti-tumor Efficacy of Pemetrexed and Altering hsa-MiR-548c-3p Expression Profile in A549 Cell Line (Human Lung Cancer)
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Abstract
Lung cancer remains a major cause of cancer-related mortality worldwide, with current treatments such as surgery, chemotherapy, and immunotherapy facing limitations, including severe side effects and high costs. Hyperthermia (H) has emerged as a promising strategy to enhance tumor sensitivity to treatments and reduce toxicity. This study investigates the effects of H in enhancing the anti-tumor efficacy of pemetrexed (PEM) and altering the expression profile of hsa-MiR-548c-3p (tumor suppressor) in the A549 cell line.
A549 cells were cultured in DMEM medium and divided into four groups: Control, and treatment with H, PEM, and a combination of H and PEM. Subsequently, cell viability, apoptosis percentage, release rate of LDH, production of ROS, and the expression level of hsa-MiR-548c-3p, CASP 8 and 9, and TYMS genes were measured.
Results revealed that the combination of H and PEM treatment had greater antitumor effects compared to the other groups. The combination of H and PEM significantly reduced cell viability, increased the percentage of apoptosis, LDH release, and ROS production, and upregulated hsa-MiR-548c-3p, CASP 8, and 9, while downregulating TYMS.
The findings suggest that H enhances the chemotherapeutic efficacy of PEM by upregulating hsa-MiR-548c-3p expression and promoting apoptosis in lung cancer cells, making it a promising complementary approach for overcoming drug resistance.
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18. Wang Z, Wu X, Hou X, Zhao W, Yang C, Wan W, et al. miR-548b-3p functions as a tumor suppressor in lung cancer. Lasers Med Sci. 2020;35(4):833-839.
19. Esmaeili Govarchin Ghaleh H, Zarei L, Mansori Motlagh B, Jabbari N. Using CuO nanoparticles and hyperthermia in radiotherapy of MCF-7 cell line: synergistic effect in cancer therapy. Artif Cells Nanomed Biotechnol. 2019;47(1):1396-1403.
20. Ghorbani Alvanegh A, Arpanaei A, Esmaeili Gouvarchin Ghaleh H, Mohammad Ganji S. MiR-320a upregulation contributes to the effectiveness of pemetrexed by inhibiting the growth and invasion of human lung cancer cell line (Calu-6). Mol Biol Rep. 2024;51(1):310.
21. Ashrafi A, Akter Z, Modareszadeh P, Modareszadeh P, Berisha E, Alemi PS, et al. Current Landscape of Therapeutic Resistance in Lung Cancer and Promising Strategies to Overcome Resistance. Cancers (Basel). 2022;14(19):4562.
22. Ribeiro TP, Moreira JA, Monteiro FJ, Laranjeira MS. Nanomaterials in cancer: Reviewing the combination of hyperthermia and triggered chemotherapy. J Control Release. 2022;347:89-103.
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24. Otmani K, Lewalle P. Tumor Suppressor miRNA in Cancer Cells and the Tumor Microenvironment: Mechanism of Deregulation and Clinical Implications. Front Oncol. 2021;11:708765.
25. Jang JH, Lee TJ. The role of microRNAs in cell death pathways. Yeungnam Univ J Med. 2021;38(2):107-117.
26. Dho SH, Cho M, Woo W, Jeong S, Kim LK. Caspases as master regulators of programmed cell death: apoptosis, pyroptosis and beyond. Exp Mol Med. 2025:1-2.
27. Li P, Zhou L, Zhao T, Liu X, Zhang P, Liu Y, et al. Caspase-9: structure, mechanisms and clinical application. Oncotarget. 2017;8(14):23996-24008.
28. Siddiqui A, Gollavilli PN, Schwab A, Vazakidou ME, Ersan PG, Ramakrishnan M, et al. Thymidylate synthase maintains the de-differentiated state of triple negative breast cancers. Cell Death Differ. 2019;26(11):2223-2236.
29. Mirra D, Esposito R, Spaziano G, La Torre C, Vocca C, Tallarico M, et al. Lung microRNAs Expression in Lung Cancer and COPD: A Preliminary Study. Biomedicines. 2023;11(3):736.
30. Carrà G, Petiti J, Tolino F, Vacca R, Orso F. MicroRNAs in metabolism for precision treatment of lung cancer. Cell Mol Biol Lett. 2024;29(1):121.
31. Ghafouri-Fard S, Aghabalazade A, Shoorei H, Majidpoor J, Taheri M, Mokhtari M. The impact of lncRNAs and miRNAs on apoptosis in lung cancer. Front oncol. 2021;11:714795.
32. Wang Z, Wu X, Hou X, Zhao W, Yang C, Wan W, et al. miR-548b-3p functions as a tumor suppressor in lung cancer. Lasers Med Sci. 2020;35(4):833-9.
33. Jin H, Zhao Y, Yang J, Zhang X, Ma S. Hyperthermia enhances the sensitivity of pancreatic cancer SW1990 cells to gemcitabine through ROS/JNK signaling. Oncol Lett. 2018;16(5):6742-6748.
34. Hwang KE, Kim YS, Hwang YR, Kwon SJ, Park DS, Cha BK, et al. Enhanced apoptosis by pemetrexed and simvastatin in malignant mesothelioma and lung cancer cells by reactive oxygen species-dependent mitochondrial dysfunction and Bim induction. Int J Oncol. 2014;45(4):1769-77.
35. Khalili E, Afgar A, Rajabpour A, Aghaee-Bakhtiari SH, Jamialahmadi K, Teimoori-Toolabi L. MiR-548c-3p through suppressing Tyms and Abcg2 increases the sensitivity of colorectal cancer cells to 5-fluorouracil. Heliyon. 2023;9(11).
2. Ettinger DS, Wood DE, Aisner DL, Akerley W, Bauman JR, Bharat A, et al. NCCN Guidelines® Insights: Non-Small Cell Lung Cancer, Version 2.2023. J Natl Compr Canc Netw. 2023;21(4):340-50.
3. Montagne F, Guisier F, Venissac N, Baste JM. The Role of Surgery in Lung Cancer Treatment: Present Indications and Future Perspectives-State of the Art. Cancers (Basel). 2021;13(15):3711.
4. de Rouw N, Piet B, Derijks HJ, van den Heuvel MM, Ter Heine R. Mechanisms, Management and Prevention of Pemetrexed-Related Toxicity. Drug Saf. 2021 Dec;44(12):1271-81.
5. Jassem J, Ramlau R, Santoro A, Schuette W, Chemaissani A, Hong S, et al. Phase III trial of pemetrexed plus best supportive care compared with best supportive care in previously treated patients with advanced malignant pleural mesothelioma. J Clin Oncol. 2008;26(10):1698-704.
6. Vicidomini G. Current Challenges and Future Advances in Lung Cancer: Genetics, Instrumental Diagnosis and Treatment. Cancers (Basel). 2023;15(14):3710.
7. Ward RA, Fawell S, Floc'h N, Flemington V, McKerrecher D, Smith PD. Challenges and Opportunities in Cancer Drug Resistance. Chem Rev. 2021 Mar 24;121(6):3297-3351.
8. Yang SJ, Huang CH, Wang CH, Shieh MJ, Chen KC. The Synergistic Effect of Hyperthermia and Chemotherapy in Magnetite Nanomedicine-Based Lung Cancer Treatment. Int J Nanomedicine. 2020;15:10331-10347.
9. Elming PB, Sørensen BS, Oei AL, Franken NAP, Crezee J, Overgaard J, et al. Hyperthermia: The Optimal Treatment to Overcome Radiation Resistant Hypoxia. Cancers (Basel). 2019;11(1):60.
10. Mantso T, Goussetis G, Franco R, Botaitis S, Pappa A, Panayiotidis M. Effects of hyperthermia as a mitigation strategy in DNA damage-based cancer therapies. Semin Cancer Biol. 2016;37-38:96-105.
11. Moradialvand M, Farashi S, Dey AD, Kumar A, Sharifi I, Hassanpour M, et al. Cell fate regulation by hyperthermia: Cancer, stem cells, and immune system. Alex Eng J. 2025;129:240-54.
12. Mao W, Li W, Hu X. Tumor hyperthermia research progress and application prospect in tumoroids (Review). Mol Clin Oncol. 2024 Feb 14;20(4):31.
13. Liebl CM, Kutschan S, Dörfler J, Käsmann L, Hübner J. Systematic review about complementary medical hyperthermia in oncology. Clin Exp Med. 2022;22(4):519-565.
14. Lutgens L, van der Zee J, Pijls-Johannesma M, De Haas-Kock DF, Buijsen J, Mastrigt GA, et al. Combined use of hyperthermia and radiation therapy for treating locally advanced cervical carcinoma. Cochrane Database Syst Rev. 2010;(1):CD006377.
15. Chakrabortty A, Patton DJ, Smith BF, Agarwal P. miRNAs: Potential as Biomarkers and Therapeutic Targets for Cancer. Genes (Basel). 2023;14(7):1375.
16. Brown C, Mantzaris M, Nicolaou E, Karanasiou G, Papageorgiou E, Curigliano G, et al. A systematic review of miRNAs as biomarkers for chemotherapy-induced cardiotoxicity in breast cancer patients reveals potentially clinically informative panels as well as key challenges in miRNA research. Cardiooncology. 2022;8(1):16.
17. Chen X, Chen Y, Hu Q, Chen X, Huang L. MiR-548c inhibits lung cancer cell proliferation through suppression of Galectin-3-mediated TLR4 signaling pathway. Int J Clin Exp Med. 2018;11(2):581-92.
18. Wang Z, Wu X, Hou X, Zhao W, Yang C, Wan W, et al. miR-548b-3p functions as a tumor suppressor in lung cancer. Lasers Med Sci. 2020;35(4):833-839.
19. Esmaeili Govarchin Ghaleh H, Zarei L, Mansori Motlagh B, Jabbari N. Using CuO nanoparticles and hyperthermia in radiotherapy of MCF-7 cell line: synergistic effect in cancer therapy. Artif Cells Nanomed Biotechnol. 2019;47(1):1396-1403.
20. Ghorbani Alvanegh A, Arpanaei A, Esmaeili Gouvarchin Ghaleh H, Mohammad Ganji S. MiR-320a upregulation contributes to the effectiveness of pemetrexed by inhibiting the growth and invasion of human lung cancer cell line (Calu-6). Mol Biol Rep. 2024;51(1):310.
21. Ashrafi A, Akter Z, Modareszadeh P, Modareszadeh P, Berisha E, Alemi PS, et al. Current Landscape of Therapeutic Resistance in Lung Cancer and Promising Strategies to Overcome Resistance. Cancers (Basel). 2022;14(19):4562.
22. Ribeiro TP, Moreira JA, Monteiro FJ, Laranjeira MS. Nanomaterials in cancer: Reviewing the combination of hyperthermia and triggered chemotherapy. J Control Release. 2022;347:89-103.
23. Bienia A, Wiecheć-Cudak O, Murzyn AA, Krzykawska-Serda M. Photodynamic Therapy and Hyperthermia in Combination Treatment-Neglected Forces in the Fight against Cancer. Pharmaceutics. 2021;13(8):1147.
24. Otmani K, Lewalle P. Tumor Suppressor miRNA in Cancer Cells and the Tumor Microenvironment: Mechanism of Deregulation and Clinical Implications. Front Oncol. 2021;11:708765.
25. Jang JH, Lee TJ. The role of microRNAs in cell death pathways. Yeungnam Univ J Med. 2021;38(2):107-117.
26. Dho SH, Cho M, Woo W, Jeong S, Kim LK. Caspases as master regulators of programmed cell death: apoptosis, pyroptosis and beyond. Exp Mol Med. 2025:1-2.
27. Li P, Zhou L, Zhao T, Liu X, Zhang P, Liu Y, et al. Caspase-9: structure, mechanisms and clinical application. Oncotarget. 2017;8(14):23996-24008.
28. Siddiqui A, Gollavilli PN, Schwab A, Vazakidou ME, Ersan PG, Ramakrishnan M, et al. Thymidylate synthase maintains the de-differentiated state of triple negative breast cancers. Cell Death Differ. 2019;26(11):2223-2236.
29. Mirra D, Esposito R, Spaziano G, La Torre C, Vocca C, Tallarico M, et al. Lung microRNAs Expression in Lung Cancer and COPD: A Preliminary Study. Biomedicines. 2023;11(3):736.
30. Carrà G, Petiti J, Tolino F, Vacca R, Orso F. MicroRNAs in metabolism for precision treatment of lung cancer. Cell Mol Biol Lett. 2024;29(1):121.
31. Ghafouri-Fard S, Aghabalazade A, Shoorei H, Majidpoor J, Taheri M, Mokhtari M. The impact of lncRNAs and miRNAs on apoptosis in lung cancer. Front oncol. 2021;11:714795.
32. Wang Z, Wu X, Hou X, Zhao W, Yang C, Wan W, et al. miR-548b-3p functions as a tumor suppressor in lung cancer. Lasers Med Sci. 2020;35(4):833-9.
33. Jin H, Zhao Y, Yang J, Zhang X, Ma S. Hyperthermia enhances the sensitivity of pancreatic cancer SW1990 cells to gemcitabine through ROS/JNK signaling. Oncol Lett. 2018;16(5):6742-6748.
34. Hwang KE, Kim YS, Hwang YR, Kwon SJ, Park DS, Cha BK, et al. Enhanced apoptosis by pemetrexed and simvastatin in malignant mesothelioma and lung cancer cells by reactive oxygen species-dependent mitochondrial dysfunction and Bim induction. Int J Oncol. 2014;45(4):1769-77.
35. Khalili E, Afgar A, Rajabpour A, Aghaee-Bakhtiari SH, Jamialahmadi K, Teimoori-Toolabi L. MiR-548c-3p through suppressing Tyms and Abcg2 increases the sensitivity of colorectal cancer cells to 5-fluorouracil. Heliyon. 2023;9(11).
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| Apoptosis hsa-MiR-548c-3p Hyperthermia Lung cancer Pemetrexed | ||
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How to Cite
1.
Mokaberi S, Babaei N, Esmaeili Gouvarchin Ghaleh H, Farnoosh G. The Role of Hyperthermia in Enhancing Anti-tumor Efficacy of Pemetrexed and Altering hsa-MiR-548c-3p Expression Profile in A549 Cell Line (Human Lung Cancer). Iran J Allergy Asthma Immunol. 2025;:1-10.
