miR-32-5p–mediated downregulation of choline kinase alpha promotes apoptosis and reduces migration in MCF7 breast cancer cells

Shaleniprieya Muniandy; Sweta Raikundalia; Ling Ling Few; Shuhaila Mat-Sharani; Get Bee Yvonne-Τee; Nor Fadhilah Kamaruzzaman; Chan Yean Yean; Wei Cun See Too; Shaleniprieya Muniandy; Sweta Raikundalia; Ling Ling Few; Shuhaila Mat-Sharani; Get Bee Yvonne-Τee; Nor Fadhilah Kamaruzzaman; Chan Yean Yean; Wei Cun See Too

doi:10.3934/molsci.2026011

AIMS Molecular Science

2026, Volume 13, Issue 2: 205-225. doi: 10.3934/molsci.2026011

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Research article

miR-32-5p–mediated downregulation of choline kinase alpha promotes apoptosis and reduces migration in MCF7 breast cancer cells

1.
School of Health Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia
2.
Nanotechnology in Veterinary Medicine Research Group, Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, 16100 Pengkalan Chepa, Kelantan, Malaysia
3.
Department of Medical Microbiology & Parasitology, School of Medical Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

Received: 04 December 2025 Revised: 05 April 2026 Accepted: 07 April 2026 Published: 21 April 2026

Choline kinase alpha (CHKA) plays an important role in phospholipid metabolism and is frequently overexpressed in several cancers, where it contributes to tumor growth and survival. In this study, we investigated whether microRNA-32-5p (miR-32-5p) regulates CHKA expression and influences malignant phenotypes in human breast cancer cells. Bioinformatic prediction and luciferase reporter assays suggested a potential interaction between miR-32-5p and the CHKA 3′ UTR. Transfection of miR-32-5p into MCF7 cells significantly reduced CHKA mRNA and CHKA protein expression. This downregulation was associated with increased apoptosis, G0/G1 cell-cycle arrest, and reduced migration of MCF7 cells. Reduced phosphorylation of ERK and mTOR was also observed, suggesting decreased activation of MAPK/mTOR signaling pathways. In contrast, although CHKA expression was reduced in non-tumorigenic MCF10A cells, apoptosis was not induced. These findings indicate that miR-32-5p regulates CHKA expression and suppresses malignant phenotypes in breast cancer cells, highlighting its potential role in modulating CHKA-associated signaling pathways.
- miR-32-5p,
- microRNAs,
- gene regulation,
- choline kinase alpha,
- breast cancer
Citation: Shaleniprieya Muniandy, Sweta Raikundalia, Ling Ling Few, Shuhaila Mat-Sharani, Get Bee Yvonne-Τee, Nor Fadhilah Kamaruzzaman, Chan Yean Yean, Wei Cun See Too. miR-32-5p–mediated downregulation of choline kinase alpha promotes apoptosis and reduces migration in MCF7 breast cancer cells[J]. AIMS Molecular Science, 2026, 13(2): 205-225. doi: 10.3934/molsci.2026011

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Abstract

Choline kinase alpha (CHKA) plays an important role in phospholipid metabolism and is frequently overexpressed in several cancers, where it contributes to tumor growth and survival. In this study, we investigated whether microRNA-32-5p (miR-32-5p) regulates CHKA expression and influences malignant phenotypes in human breast cancer cells. Bioinformatic prediction and luciferase reporter assays suggested a potential interaction between miR-32-5p and the CHKA 3′ UTR. Transfection of miR-32-5p into MCF7 cells significantly reduced CHKA mRNA and CHKA protein expression. This downregulation was associated with increased apoptosis, G0/G1 cell-cycle arrest, and reduced migration of MCF7 cells. Reduced phosphorylation of ERK and mTOR was also observed, suggesting decreased activation of MAPK/mTOR signaling pathways. In contrast, although CHKA expression was reduced in non-tumorigenic MCF10A cells, apoptosis was not induced. These findings indicate that miR-32-5p regulates CHKA expression and suppresses malignant phenotypes in breast cancer cells, highlighting its potential role in modulating CHKA-associated signaling pathways.

Acknowledgments

This research was funded by the Universiti Sains Malaysia Bridging Grant with Project No: R501-LR-RND003-0000002377-0000. We gratefully acknowledge the technical support provided by the laboratory personnel of the School of Health Sciences, USM, as well as the staff of the Central Research Laboratory, School of Medical Sciences, USM.

Conflict of interest

The authors declare that they have no competing interests.

References

[1]	Arlauckas SP, Popov AV, Delikatny EJ (2016) Choline kinase alpha—Putting the ChoK-hold on tumor metabolism. Prog Lipid Res 63: 28-40. https://doi.org/10.1016/j.plipres.2016.03.005
[2]	Chang CC, Few LL, Konrad M, et al. (2016) Phosphorylation of human choline kinase Beta by protein kinase A: Its impact on activity and inhibition. PLoS One 11: e0154702. https://doi.org/10.1371/journal.pone.0154702
[3]	Lacal JC, Zimmerman T, Campos JM (2021) Choline kinase: An unexpected journey for a precision medicine strategy in human diseases. Pharmaceutics 13: 788. https://doi.org/10.3390/pharmaceutics13060788
[4]	Korbecki J, Bosiacki M, Kupnicka P, et al. (2025) Choline kinases: Enzyme activity, involvement in cancer and other diseases, inhibitors. Int J Cancer 156: 1314-1325. https://doi.org/10.1002/ijc.35286
[5]	Glunde K, Bhujwalla ZM, Ronen SM (2011) Choline metabolism in malignant transformation. Nat Rev Cancer 11: 835-848. https://doi.org/10.1038/nrc3162
[6]	de Molina AR, Rodríguez-González A, Gutiérrez R, et al. (2002) Overexpression of choline kinase is a frequent feature in human tumor-derived cell lines and in lung, prostate, and colorectal human cancers. Biochem Biophys Res Commun 296: 580-583. https://doi.org/10.1016/s0006-291x(02)00920-8
[7]	Yalcin A, Clem B, Makoni S, et al. (2010) Selective inhibition of choline kinase simultaneously attenuates MAPK and PI3K/AKT signaling. Oncogene 29: 139-149. https://doi.org/10.1038/onc.2009.317
[8]	Clem BF, Clem AL, Yalcin A, et al. (2011) A novel small molecule antagonist of choline kinase-α that simultaneously suppresses MAPK and PI3K/AKT signaling. Oncogene 30: 3370-3380. https://doi.org/10.1038/onc.2011.51
[9]	Chua BT, Gallego-Ortega D, de Molina AR, et al. (2009) Regulation of Akt(ser473) phosphorylation by Choline kinase in breast carcinoma cells. Mol Cancer 8: 131. https://doi.org/10.1186/1476-4598-8-131
[10]	Lin XM, Hu L, Gu J, et al. (2017) Choline kinase α mediates interactions between the epidermal growth factor receptor and mechanistic target of rapamycin complex 2 in hepatocellular carcinoma cells to promote drug resistance and xenograft tumor progression. Gastroenterology 152: 1187-1202. https://doi.org/10.1053/j.gastro.2016.12.033
[11]	Mariotto E, Bortolozzi R, Volpin I, et al. (2018) EB-3D a novel choline kinase inhibitor induces deregulation of the AMPK-mTOR pathway and apoptosis in leukemia T-cells. Biochem Pharmacol 155: 213-223. https://doi.org/10.1016/j.bcp.2018.07.004
[12]	Gokhale S, Xie P (2021) ChoK-full of potential: Choline kinase in B cell and T cell malignancies. Pharmaceutics 13: 911. https://doi.org/10.3390/pharmaceutics13060911
[13]	Gruber J, See Too WC, Wong MT, et al. (2012) Balance of human choline kinase isoforms is critical for cell cycle regulation. FEBS J 279: 1915-1928. https://doi.org/10.1111/j.1742-4658.2012.08573.x
[14]	Wu G, Vance DE (2010) Choline kinase and its function. Biochem Cell Biol 88: 559-564. https://doi.org/10.1139/O09-160
[15]	Schiaffino-Ortega S, Baglioni E, Mariotto E, et al. (2016) Design, synthesis, crystallization and biological evaluation of new symmetrical biscationic compounds as selective inhibitors of human Choline Kinase α1 (ChoKα1). Sci Rep 6: 23793. https://doi.org/10.1038/srep23793
[16]	Ayub Khan SM, Few LL, See Too WC (2018) Downregulation of human choline kinase α gene expression by miR-876-5p. Mol Med Rep 17: 7442-7450. https://doi.org/10.3892/mmr.2018.8762
[17]	Raikundalia S, Sa'Dom SAFM, Few LL, et al. (2021) MicroRNA-367-3p induces apoptosis and suppresses migration of MCF-7 cells by downregulating the expression of human choline kinase α. Oncol Lett 21: 183. https://doi.org/10.3892/ol.2021.12444
[18]	Raikundalia S, Few LL, Hassan SA, et al. (2024) Choline kinase and miR-32-5p: A crucial interaction promoting apoptosis and delaying wound repair in cervical cancer cells. AIMS Biophys 11: 281-295. https://doi.org/10.3934/biophy.2024016
[19]	Rodríguez-González A, de Molina AR, Bañez-Coronel M, et al. (2005) Inhibition of choline kinase renders a highly selective cytotoxic effect in tumour cells through a mitochondrial independent mechanism. Int J Oncol 26: 999-1008.
[20]	Bañez-Coronel M, de Molina AR, Rodríguez-González A, et al. (2008) Choline kinase alpha depletion selectively kills tumoral cells. Curr Cancer Drug Targets 8: 709-719. https://doi.org/10.2174/156800908786733432
[21]	Gebert LFR, MacRae IJ (2019) Regulation of microRNA function in animals. Nat Rev Mol Cell Biol 20: 21-37. https://doi.org/10.1038/s41580-018-0045-7
[22]	Hirschberger S, Hinske LC, Kreth S (2018) MiRNAs: Dynamic regulators of immune cell functions in inflammation and cancer. Cancer Lett 431: 11-21. https://doi.org/10.1016/j.canlet.2018.05.020
[23]	Lan H, Lu H, Wang X, et al. (2015) MicroRNAs as potential biomarkers in cancer: opportunities and challenges. Biomed Res Int 2015: 125094. https://doi.org/10.1155/2015/125094
[24]	Shah V, Shah J (2020) Recent trends in targeting miRNAs for cancer therapy. J Pharm Pharmacol 72: 1732-1749. https://doi.org/10.1111/jphp.13351
[25]	Mayuri K, Vickram S, Anand T, et al. (2025) MicroRNA-mediated regulation of BCL-2 in breast cancer. AIMS Mol Sci 12: 32-48. https://doi.org/10.3934/molsci.2025003
[26]	Yuan P, Tang C, Chen B, et al. (2021) miR-32-5p suppresses the proliferation and migration of pancreatic adenocarcinoma cells by targeting TLDC1. Mol Med Rep 24: 752. https://doi.org/10.3892/mmr.2021.12392
[27]	Sun C, Huang LG, Leng B, et al. (2025) MicroRNA-32-5p promotes the proliferation and metastasis of gastric cancer cells. Sci Rep 15: 2282. https://doi.org/10.1038/s41598-025-86367-3
[28]	Xia W, Zhou J, Luo H, et al. (2017) MicroRNA-32 promotes cell proliferation, migration and suppresses apoptosis in breast cancer cells by targeting FBXW7. Cancer Cell Int 17: 14. https://doi.org/10.1186/s12935-017-0383-0
[29]	McGeary SE, Lin KS, Shi CY, et al. (2019) The biochemical basis of microRNA targeting efficacy. Science 366: eaav1741. https://doi.org/10.1126/science.aav1741
[30]	Brennecke J, Stark A, Russell RB, et al. (2005) Principles of microRNA-target recognition. PLoS Biol 3: e85. https://doi.org/10.1371/journal.pbio.0030085
[31]	Grimson A, Farh KK-H, Johnston WK, et al. (2007) MicroRNA targeting specificity in mammals: determinants beyond seed pairing. Mol Cell 27: 91-105. https://doi.org/10.1016/j.molcel.2007.06.017
[32]	Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCt method. Methods 25: 402-408. https://doi.org/10.1006/meth.2001.1262
[33]	Shen Z, Xue D, Wang K, et al. (2022) Metformin exerts an antitumor effect by inhibiting bladder cancer cell migration and growth, and promoting apoptosis through the PI3K/AKT/mTOR pathway. BMC Urol 22: 79. https://doi.org/10.1186/s12894-022-01027-2
[34]	Yue J, López JM (2020) Understanding MAPK signaling pathways in apoptosis. Int J Mol Sci 21. https://doi.org/10.3390/ijms21072346
[35]	Wang N, Brickute D, Braga M, et al. (2021) Novel non-congeneric derivatives of the choline kinase Alpha inhibitor ICL-CCIC-0019. Pharmaceutics 13: 1078. https://doi.org/10.3390/pharmaceutics13071078
[36]	de Molina AR, Gutiérrez R, Ramos MA, et al. (2002) Increased choline kinase activity in human breast carcinomas: Clinical evidence for a potential novel antitumor strategy. Oncogene 21: 4317-4322. https://doi.org/10.1038/sj.onc.1205556
[37]	Penet MF, Shah T, Bharti S, et al. (2015) Metabolic imaging of pancreatic ductal adenocarcinoma detects altered choline metabolism. Clin Cancer Res 21: 386-395. https://doi.org/10.1158/1078-0432.CCR-14-0964
[38]	Iorio E, Ricci A, Bagnoli M, et al. (2010) Activation of phosphatidylcholine cycle enzymes in human epithelial ovarian cancer cells. Cancer Res 70: 2126-2135. https://doi.org/10.1158/0008-5472.CAN-09-3833
[39]	Trousil S, Lee P, Pinato DJ, et al. (2014) Alterations of choline phospholipid metabolism in endometrial cancer are caused by choline kinase alpha overexpression and a hyperactivated deacylation pathway. Cancer Res 74: 6867-6877. https://doi.org/10.1158/0008-5472.CAN-13-2409
[40]	de Molina AR, Sarmentero-Estrada J, Belda-Iniesta C, et al. (2007) Expression of choline kinase alpha to predict outcome in patients with early-stage non-small-cell lung cancer: A retrospective study. Lancet Oncol 8: 889-897. https://doi.org/10.1016/S1470-2045(07)70279-6
[41]	Chen Z, Krishnamachary B, Bhujwalla ZM (2016) Degradable dextran nanopolymer as a carrier for choline kinase (ChoK) siRNA cancer therapy. Nanomaterials 6: 34. https://doi.org/10.3390/nano6020034
[42]	Chen H, Xie G, Luo Q, et al. (2023) Regulatory miRNAs, circRNAs and lncRNAs in cell cycle progression of breast cancer. Funct Integr Genomics 23: 233. https://doi.org/10.1007/s10142-023-01130-z
[43]	Darvish L, Bahreyni Toossi MT, Azimian H, et al. (2023) The role of microRNA-induced apoptosis in diverse radioresistant cancers. Cell Signal 104: 110580. https://doi.org/10.1016/j.cellsig.2022.110580
[44]	Ma L (2016) MicroRNA and metastasis. Advances in cancer research 132: 165-207. https://doi.org/10.1016/bs.acr.2016.07.004
[45]	Petri BJ, Klinge CM (2020) Regulation of breast cancer metastasis signaling by miRNAs. Cancer Metastasis Rev 39: 837-886. https://doi.org/10.1007/s10555-020-09905-7
[46]	Oliveira AC, Bovolenta LA, Nachtigall PG, et al. (2017) Combining results from distinct MicroRNA target prediction tools enhances the performance of analyses. Front Genet 8: 59. https://doi.org/10.3389/fgene.2017.00059
[47]	Zheng Z, Reichel M, Deveson I, et al. (2017) Target RNA secondary structure is a major determinant of miR159 efficacy. Plant Physiol 174: 1764-1778. https://doi.org/10.1104/pp.16.01898
[48]	Kang T, Sun WL, Lu XF, et al. (2020) MiR-28-5p mediates the anti-proliferative and pro-apoptotic effects of curcumin on human diffuse large B-cell lymphoma cells. J Int Med Res 48: 1-13. https://doi.org/10.1177/0300060520943792
[49]	Li JX, Li Y, Xia T, et al. (2021) miR-21 Exerts Anti-proliferative and Pro-apoptotic Effects in LPS-induced WI-38 Cells via Directly Targeting TIMP3. Cell Biochem Biophys 79: 781-790. https://doi.org/10.1007/s12013-021-00987-w
[50]	Yang J, Niu H, Chen X (2021) GATA1-activated HNF1A-AS1 facilitates the progression of triple-negative breast cancer via sponging miR-32-5p to upregulate RNF38. Cancer Manag Res 2021: 1357-1369. https://doi.org/10.2147/CMAR.S274204
[51]	Qin SY, Li B, Chen M, et al. (2022) MiR-32-5p promoted epithelial-to-mesenchymal transition of oral squamous cell carcinoma cells via regulating the KLF2/CXCR4 pathway. Kaohsiung J Med Sci 38: 120-128. https://doi.org/10.1002/kjm2.12450
[52]	Zeng S, Liu S, Feng J, et al. (2020) MicroRNA-32 promotes ovarian cancer cell proliferation and motility by targeting SMG1. Oncol Lett 20: 733-741. https://doi.org/10.3892/ol.2020.11624
[53]	Zhang JX, Yang W, Wu JZ, et al. (2021) MicroRNA-32-5p inhibits epithelial-mesenchymal transition and metastasis in lung adenocarcinoma by targeting SMAD family 3. J Cancer 12: 2258-2267. https://doi.org/10.7150/jca.48387
[54]	Liu YJ, Zhou HG, Chen LH, et al. (2019) MiR-32-5p regulates the proliferation and metastasis of cervical cancer cells by targeting HOXB8. Eur Rev Med Pharmacol Sci 23: 87-95. https://doi.org/10.26355/eurrev_201901_16752
[55]	Ni F, Gui Z, Guo Q, et al. (2016) Downregulation of miR-362-5p inhibits proliferation, migration and invasion of human breast cancer MCF7 cells. Oncol Lett 11: 1155-1160. https://doi.org/10.3892/ol.2015.3993

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