MicroRNA‑15a promotes prostate cancer cell ferroptosis by inhibiting GPX4 expression

Xu,Po; Wang,Ying; Deng,Zhe; Tan,Zhibo; Pei,Xiaojuan

doi:10.3892/ol.2022.13186

MicroRNA‑15a promotes prostate cancer cell ferroptosis by inhibiting GPX4 expression

Authors:
- Po Xu
- Ying Wang
- Zhe Deng
- Zhibo Tan
- Xiaojuan Pei
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Affiliations: Department of Emergency, The First Affiliated Hospital, Shenzhen University, Shenzhen Second People's Hospital, Shenzhen, Guangdong 518000, P.R. China, Medical Oncology Ward 1, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital and Shenzhen Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Shenzhen, Guangdong 518116, P.R. China, Department of Radiation Oncology, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518100, P.R. China, Department of Pathology, Shenzhen Hospital, Southern Medical University, Shenzhen, Guangdong 518100, P.R. China
Published online on: January 3, 2022 https://doi.org/10.3892/ol.2022.13186
Article Number: 67

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Abstract

Ferroptosis is a novel form of regulated cell death characterized by accumulated lipid reactive oxygen species (ROS) and inactivation of glutathione peroxidase 4 (GPX4). The present study aimed to investigate the role of microRNA (miRNA/miR)‑15a in ferroptosis of prostate cancer cells. Bioinformatics analysis was performed to predict the potential interaction between miR‑15a and the 3'‑untranslated region (UTR) of GPX4 mRNA. The prostate cancer cell line, LNCAP was transfected with miR‑15a mimics or small interfering (si)‑GPX4. Reverse transcription‑quantitative PCR and western blot analyses were performed to detect the mRNA and protein expression levels of GPX4, respectively. Biotin‑RNA pull‑down and dual‑luciferase reporter assays were performed to verify the interaction between miR‑15a and GPX4 mRNA. The Cell Counting Kit‑8 assay was performed to assess cell proliferation, while lactate dehydrogenase (LDH) and intracellular ferrous iron levels were detected via ELISA. Lipid ROS and mitochondrial membrane potential (MMP) were assessed via flow cytometry and staining with C11‑BIODIPY probes or JC‑1. Furthermore, lipid peroxidation was identified by measuring malondialdehyde (MDA) levels. The results demonstrated that transfection with miR‑15a mimics decreased GPX4 protein expression. Bioinformatics analysis revealed potential binding sites between miR‑15a and the 3'‑UTR region of GPX4, and RNA pull‑down and the dual‑luciferase reporter assays further confirmed the interaction between miR‑15a and GPX4 mRNA. Both transfection with miR‑15a mimics and si‑GPX4 suppressed cell proliferation, elevated LDH release, accumulated intracellular ferrous iron and ROS, disrupted MMP and increased MDA levels. Taken together, the results of the present study suggest miR‑15a induces ferroptosis by regulating GPX4 in prostate cancer cells, which provides evidence for investigating the therapeutic strategies of prostate cancer.

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1	Omar MI, Roobol MJ, Ribal MJ, Abbott T, Agapow PM, Araujo S, Asiimwe A, Auffray C, Balaur I, Beyer K, et al: Introducing PIONEER: A project to harness big data in prostate cancer research. Nat Rev Urol. 17:351–362. 2020. View Article : Google Scholar : PubMed/NCBI
2	Hu C, Xia H, Bai S, Zhao J, Edwards H, Li X, Yang Y, Lyu J, Wang G, Zhan Y, et al: CUDC-907, a novel dual PI3K and HDAC inhibitor, in prostate cancer: Antitumour activity and molecular mechanism of action. J Cell Mol Med. 24:7239–7253. 2020. View Article : Google Scholar : PubMed/NCBI
3	Hassannia B, Vandenabeele P and Vanden Berghe T: Targeting ferroptosis to iron out cancer. Cancer Cell. 35:830–849. 2019. View Article : Google Scholar : PubMed/NCBI
4	Hao S, Yu J, He W, Huang Q, Zhao Y, Liang B, Zhang S, Wen Z, Dong S, Rao J, et al: Cysteine dioxygenase 1 mediates erastin-induced ferroptosis in human gastric cancer cells. Neoplasia. 19:1022–1032. 2017. View Article : Google Scholar : PubMed/NCBI
5	Zhang Y, Shi J, Liu X, Feng L, Gong Z, Koppula P, Sirohi K, Li X, Wei Y, Lee H, et al: BAP1 links metabolic regulation of ferroptosis to tumour suppression. Nat Cell Biol. 20:1181–1192. 2018. View Article : Google Scholar : PubMed/NCBI
6	Yang WS, SriRamaratnam R, Welsch ME, Shimada K, Skouta R, Viswanathan VS, Cheah JH, Clemons PA, Shamji AF, Clish CB, et al: Regulation of ferroptotic cancer cell death by GPX4. Cell. 156:317–331. 2014. View Article : Google Scholar : PubMed/NCBI
7	Seibt TM, Proneth B and Conrad M: Role of GPX4 in ferroptosis and its pharmacological implication. Free Radic Biol Med. 133:144–152. 2019. View Article : Google Scholar : PubMed/NCBI
8	Louandre C, Marcq I, Bouhlal H, Lachaier E, Godin C, Saidak Z, François C, Chatelain D, Debuysscher V, Barbare JC, et al: The retinoblastoma (Rb) protein regulates ferroptosis induced by sorafenib in human hepatocellular carcinoma cells. Cancer Lett. 356:971–977. 2015. View Article : Google Scholar : PubMed/NCBI
9	Sui X, Zhang R, Liu S, Duan T, Zhai L, Zhang M, Han X, Xiang Y, Huang X, Lin H and Xie T: RSL3 drives ferroptosis through GPX4 Inactivation and ROS production in colorectal cancer. Front Pharmacol. 9:13712018. View Article : Google Scholar : PubMed/NCBI
10	Zou Y, Palte MJ, Deik AA, Li H, Eaton JK, Wang W, Tseng YY, Deasy R, Kost-Alimova M, Dančík V, et al: A GPX4-dependent cancer cell state underlies the clear-cell morphology and confers sensitivity to ferroptosis. Nat Commun. 10:16172019. View Article : Google Scholar : PubMed/NCBI
11	Baldassarri M, Fallerini C, Cetta F, Ghisalberti M, Bellan C, Furini S, Spiga O, Crispino S, Gotti G, Ariani F, et al: Omic approach in non-smoker female with lung squamous cell carcinoma pinpoints to germline susceptibility and personalized medicine. Cancer Res Treat. 50:356–365. 2018. View Article : Google Scholar : PubMed/NCBI
12	Karbasforooshan H, Roohbakhsh A and Karimi G: SIRT1 and microRNAs: The role in breast, lung and prostate cancers. Exp Cell Res. 367:1–6. 2018. View Article : Google Scholar : PubMed/NCBI
13	Sun Z, Shi K, Yang S, Liu J, Zhou Q, Wang G, Song J, Li Z, Zhang Z and Yuan W: Effect of exosomal miRNA on cancer biology and clinical applications. Mol Cancer. 17:1472018. View Article : Google Scholar : PubMed/NCBI
14	Rupaimoole R and Slack FJ: MicroRNA therapeutics: Towards a new era for the management of cancer and other diseases. Nat Rev Drug Discov. 16:203–222. 2017. View Article : Google Scholar : PubMed/NCBI
15	Guelfi G, Cochetti G, Stefanetti V, Zampini D, Diverio S, Boni A and Mearini E: Next Generation Sequencing of urine exfoliated cells: An approach of prostate cancer microRNAs research. Sci Rep. 8:71112018. View Article : Google Scholar : PubMed/NCBI
16	Zhang K, Wu L, Zhang P, Luo M, Du J, Gao T, O'Connell D, Wang G, Wang H and Yang Y: miR-9 regulates ferroptosis by targeting glutamic-oxaloacetic transaminase GOT1 in melanoma. Mol Carcinog. 57:1566–1576. 2018. View Article : Google Scholar : PubMed/NCBI
17	Luo M, Wu L, Zhang K, Wang H, Zhang T, Gutierrez L, O'Connell D, Zhang P, Li Y, Gao T, et al: miR-137 regulates ferroptosis by targeting glutamine transporter SLC1A5 in melanoma. Cell Death Differ. 25:1457–1472. 2018. View Article : Google Scholar : PubMed/NCBI
18	Tomita K, Fukumoto M, Itoh K, Kuwahara Y, Igarashi K, Nagasawa T, Suzuki M, Kurimasa A and Sato T: MiR-7-5p is a key factor that controls radioresistance via intracellular Fe²⁺ content in clinically relevant radioresistant cells. Biochem Biophys Res Commun. 518:712–718. 2019. View Article : Google Scholar : PubMed/NCBI
19	Zhang H, Deng T, Liu R, Ning T, Yang H, Liu D, Zhang Q, Lin D, Ge S, Bai M, et al: CAF secreted miR-522 suppresses ferroptosis and promotes acquired chemo-resistance in gastric cancer. Mol Cancer. 19:432020. View Article : Google Scholar : PubMed/NCBI
20	Cui Y, Yang Y, Ren L, Yang J, Wang B, Xing T, Chen H and Chen M: miR-15a-3p suppresses prostate cancer cell proliferation and invasion by targeting SLC39A7 via downregulating Wnt/β-catenin signaling pathway. Cancer Biother Radiopharm. 34:472–479. 2019. View Article : Google Scholar : PubMed/NCBI
21	Jia X, Liu H, Xu C, Han S, Shen Y, Miao X, Hu X, Lin Z, Qian L, Wang Z and Gong W: MiR-15a/16-1 deficiency induces IL-10-producing CD19⁺ TIM-1⁺ cells in tumor microenvironment. J Cell Mol Med. 23:1343–1353. 2019. View Article : Google Scholar : PubMed/NCBI
22	Al-Kafaji G, Said HM, Alam MA and Al Naieb ZT: Blood-based microRNAs as diagnostic biomarkers to discriminate localized prostate cancer from benign prostatic hyperplasia and allow cancer-risk stratification. Oncol Lett. 16:1357–1365. 2018.PubMed/NCBI
23	Jin W, Chen F, Wang K, Song Y, Fei X and Wu B: miR-15a/miR-16 cluster inhibits invasion of prostate cancer cells by suppressing TGF-β signaling pathway. Biomed Pharmacother. 104:637–644. 2018. View Article : Google Scholar : PubMed/NCBI
24	Bonci D, Coppola V, Musumeci M, Addario A, Giuffrida R, Memeo L, D'Urso L, Pagliuca A, Biffoni M, Labbaye C, et al: The miR-15a-miR-16-1 cluster controls prostate cancer by targeting multiple oncogenic activities. Nat Med. 14:1271–1277. 2008. View Article : Google Scholar : PubMed/NCBI
25	Shimomura A, Shiino S, Kawauchi J, Takizawa S, Sakamoto H, Matsuzaki J, Ono M, Takeshita F, Niida S, Shimizu C, et al: Novel combination of serum microRNA for detecting breast cancer in the early stage. Cancer Sci. 107:326–334. 2016. View Article : Google Scholar : PubMed/NCBI
26	Qadir MI and Faheem A: miRNA: A diagnostic and therapeutic tool for pancreatic cancer. Crit Rev Eukaryot Gene Expr. 27:197–204. 2017. View Article : Google Scholar : PubMed/NCBI
27	Yokoi A, Matsuzaki J, Yamamoto Y, Yoneoka Y, Takahashi K, Shimizu H, Uehara T, Ishikawa M, Ikeda SI, Sonoda T, et al: Integrated extracellular microRNA profiling for ovarian cancer screening. Nat Commun. 9:43192018. View Article : Google Scholar : PubMed/NCBI
28	Omidkhoda N, Wallace Hayes A, Reiter RJ and Karimi G: The role of MicroRNAs on endoplasmic reticulum stress in myocardial ischemia and cardiac hypertrophy. Pharmacol Res. 150:1045162019. View Article : Google Scholar : PubMed/NCBI
29	Jiang J, Jia P, Zhao Z and Shen B: Key regulators in prostate cancer identified by co-expression module analysis. BMC Genomics. 15:10152014. View Article : Google Scholar : PubMed/NCBI
30	Hart M, Nolte E, Wach S, Szczyrba J, Taubert H, Rau TT, Hartmann A, Grässer FA and Wullich B: Comparative microRNA profiling of prostate carcinomas with increasing tumor stage by deep sequencing. Mol Cancer Res. 12:250–263. 2014. View Article : Google Scholar : PubMed/NCBI
31	Leite KR, Tomiyama A, Reis ST, Sousa-Canavez JM, Sañudo A, Camara-Lopes LH and Srougi M: MicroRNA expression profiles in the progression of prostate cancer-from high-grade prostate intraepithelial neoplasia to metastasis. Urol Oncol. 31:796–801. 2013. View Article : Google Scholar : PubMed/NCBI
32	Zidan HE, Abdul-Maksoud RS, Elsayed WSH and Desoky EAM: Diagnostic and prognostic value of serum miR-15a and miR-16-1 expression among egyptian patients with prostate cancer. IUBMB Life. 70:437–444. 2018. View Article : Google Scholar : PubMed/NCBI
33	Aqeilan RI, Calin GA and Croce CM: miR-15a and miR-16-1 in cancer: Discovery, function and future perspectives. Cell Death Differ. 17:215–220. 2010. View Article : Google Scholar : PubMed/NCBI
34	Antognelli C, Mezzasoma L, Fettucciari K, Mearini E and Talesa VN: Role of glyoxalase I in the proliferation and apoptosis control of human LNCaP and PC3 prostate cancer cells. Prostate. 73:121–132. 2013. View Article : Google Scholar : PubMed/NCBI
35	Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, Patel DN, Bauer AJ, Cantley AM, Yang WS, et al: Ferroptosis: An iron-dependent form of nonapoptotic cell death. Cell. 149:1060–1072. 2012. View Article : Google Scholar : PubMed/NCBI
36	Yu M, Gai C, Li Z, Ding D, Zheng J, Zhang W, Lv S and Li W: Targeted exosome-encapsulated erastin induced ferroptosis in triple negative breast cancer cells. Cancer Sci. 110:3173–3182. 2019. View Article : Google Scholar : PubMed/NCBI
37	Shin D, Kim EH, Lee J and Roh JL: Nrf2 inhibition reverses resistance to GPX4 inhibitor-induced ferroptosis in head and neck cancer. Free Radic Biol Med. 129:454–462. 2018. View Article : Google Scholar : PubMed/NCBI
38	Bebber CM, Müller F, Prieto Clemente L, Weber J and von Karstedt S: Ferroptosis in cancer cell biology. Cancers (Basel). 12:1642020. View Article : Google Scholar : PubMed/NCBI