MicroRNA‑195‑5p suppresses the proliferation, migration, invasion and epithelial‑mesenchymal transition of laryngeal cancer cells <em>in vitro</em> by targeting E2F3

Zhou,Min; Wang,Yu; Zhang,Changming; Qi,Meihao; Yao,Min; Sun,Lizhi; Xu,Xining

doi:10.3892/etm.2021.10512

MicroRNA‑195‑5p suppresses the proliferation, migration, invasion and epithelial‑mesenchymal transition of laryngeal cancer cells in vitro by targeting E2F3

Authors:
- Min Zhou
- Yu Wang
- Changming Zhang
- Meihao Qi
- Min Yao
- Lizhi Sun
- Xining Xu
View Affiliations

Affiliations: Department of Otolaryngology Head and Neck Surgery, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Operating Room, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Otolaryngology, The 940th Hospital of The Joint Logistic Support Force of PLA, Lanzhou, Gansu 730050, P.R. China
Published online on: July 28, 2021 https://doi.org/10.3892/etm.2021.10512
Article Number: 1078
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Increasing evidence has indicated that microRNAs (miRNAs/miRs) play an important role in the occurrence and development of various types of cancer. The aim of the present study was to investigate the role and underlying molecular mechanisms of miR‑195‑5p in laryngeal cancer cell proliferation, migration and invasion. Reverse transcription‑quantitative PCR (RT‑qPCR) was performed to measure the expression levels of miR‑195‑5p in laryngeal carcinoma cell lines. The expression levels of miR‑195‑5p and E2F transcription factor 3 (E2F3) were modified by transfection with miR‑195‑5p mimics and pcDNA3.1‑E2F3. A luciferase reporter assay was used to verify the association between miR‑195a‑5p and E2F3. Cell Counting Kit‑8, cell wound healing and Transwell invasion assays were used to detect the biological functions of laryngeal cancer cells. The expression of epithelial‑mesenchymal transition (EMT)‑associated genes was evaluated by western blotting and RT‑qPCR. The results revealed that the expression of miR‑195‑5p was decreased in laryngeal cancer cell lines. The overexpression of miR‑195‑5p inhibited the proliferation, migration, invasion and EMT of laryngeal cancer cells. Dual‑luciferase reporter assays revealed that miR‑195‑5p could directly target E2F3 and that there was a negative association between them. E2F3 overexpression significantly attenuated the inhibitory effects of the overexpression of miR‑195‑5p on the proliferation, migration, invasion and EMT of laryngeal cancer cells. Collectively, the findings of the present study demonstrated that the overexpression of miR‑195‑5p significantly inhibited the progression of laryngeal cancer cells, and these effects may be mediated via the downregulation of the expression of E2F3.

View Figures

View References

1	Thompson LD: Laryngeal dysplasia, squamous cell carcinoma, and variants. Surg Pathol Clin. 10:15–33. 2017.PubMed/NCBI View Article : Google Scholar
2	Echanique KA, Desai SV, Marchiano E, Spinazzi EF, Strojan P, Baredes S and Eloy JA: Laryngeal verrucous carcinoma: A systematic review. Otolaryngol Head Neck Surg. 156:38–45. 2017.PubMed/NCBI View Article : Google Scholar
3	Kolator M, Kolator P and Zatoński T: Assessment of quality of life in patients with laryngeal cancer: A review of articles. Adv Clin Exp Med. 27:711–715. 2018.PubMed/NCBI View Article : Google Scholar
4	Feng W, Yang Y, Xin ZH and Ni XG: Misdiagnosis experience of laryngeal carcinoma with acute laryngitis as the first symptom. Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 53(640)2018.PubMed/NCBI View Article : Google Scholar : (In Chinese).
5	Obid R, Redlich M and Tomeh C: The treatment of laryngeal cancer. Oral Maxillofac Surg Clin North Am. 31:1–11. 2019.PubMed/NCBI View Article : Google Scholar
6	Chang HY, Lee CH, Li YS, Huang JT, Lan SH, Wang YF, Lai WW, Wang YC, Lin YJ, Liu HS and Cheng HC: MicroRNA-146a suppresses tumor malignancy via targeting vimentin in esophageal squamous cell carcinoma cells with lower fibronectin membrane assembly. J Biomed Sci. 27(102)2020.PubMed/NCBI View Article : Google Scholar
7	Escuin D, López-Vilaró L, Bell O, Mora J, Moral A, Pérez JI, Arqueros C, Ramón Y, Cajal T, Lerma E and Barnadas A: MicroRNA-1291 is associated with locoregional metastases in patients with early-stage breast cancer. Front Genet. 11(562114)2020.PubMed/NCBI View Article : Google Scholar
8	Xu WG, Kuang YM, Wang D, Li Z and Xia RP: Effect of miR-210 on proliferation and migration of pancreatic cancer cells through regulating Runx3 level. J Biomater Tissue Eng. 10:1827–1831. 2020.
9	Chen L, Sun DZ, Fu YG, Yang PZ, Lv HQ, Gao Y and Zhang XY: Upregulation of microRNA-141 suppresses epithelial-mesenchymal transition and lymph node metastasis in laryngeal cancer through HOXC6-dependent TGF-β signaling pathway. Cell Signal. 66(109444)2020.PubMed/NCBI View Article : Google Scholar
10	Yang B, Zang J, Yuan W, Jiang X and Zhang F: The miR-136-5p/ROCK1 axis suppresses invasion and migration, and enhances cisplatin sensitivity in head and neck cancer cells. Exp Ther Med. 21(317)2021.PubMed/NCBI View Article : Google Scholar
11	Liu YB, Wang Y, Zhang MD, Yue W and Sun CN: MicroRNA-29a functions as a tumor suppressor through targeting STAT3 in laryngeal squamous cell carcinoma. Exp Mol Pathol. 116(104521)2020.PubMed/NCBI View Article : Google Scholar
12	Cossu AM, Mosca L, Zappavigna S, Misso G, Bocchetti M, De Micco F, Quagliuolo L, Porcelli M, Caraglia M and Boccellino M: Long non-coding RNAs as important biomarkers in laryngeal cancer and other head and neck tumours. Int J Mol Sci. 20(3444)2019.PubMed/NCBI View Article : Google Scholar
13	Erkul E, Yilmaz I, Gungor A, Kurt O and Babayigit MA: MicroRNA-21 in laryngeal squamous cell carcinoma: Diagnostic and prognostic features. Laryngoscope. 127:E62–E66. 2017.PubMed/NCBI View Article : Google Scholar
14	Li W, Ma HP and Sun J: MicroRNA-34a/c function as tumor suppressors in Hep-2 laryngeal carcinoma cells and may reduce GALNT7 expression. Mol Med Rep. 9:1293–1298. 2014.PubMed/NCBI View Article : Google Scholar
15	Guo Y, An R, Zhao R, Sun Y, Liu M and Tian L: miR-375 exhibits a more effective tumor-suppressor function in laryngeal squamous carcinoma cells by regulating KLF4 expression compared with simple co-transfection of miR-375 and miR-206. Oncol Rep. 36:952–960. 2016.PubMed/NCBI View Article : Google Scholar
16	Chai L, Kang XJ, Sun ZZ, Zeng MF, Yu SR, Ding Y, Liang JQ, Li TT and Zhao J: MiR-497-5p, miR-195-5p and miR-455-3p function as tumor suppressors by targeting hTERT in melanoma A375 cells. Cancer Manag Res. 10:989–1003. 2018.PubMed/NCBI View Article : Google Scholar
17	Feng C, Zhang L, Sun Y, Li X, Zhan L, Lou Y, Wang Y, Liu L and Zhang Y: GDPD5, a target of miR-195-5p, is associated with metastasis and chemoresistance in colorectal cancer. Biomed Pharmacother. 101:945–952. 2018.PubMed/NCBI View Article : Google Scholar
18	Liu X, Zhou Y, Ning YE, Gu H, Tong Y and Wang N: MiR-195-5p inhibits malignant progression of cervical cancer by targeting YAP1. OncoTargets Ther. 13:931–944. 2020.PubMed/NCBI View Article : Google Scholar
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
20	Poel D, Boyd LNC, Beekhof R, Schelfhorst T, Pham TV, Piersma SR, Knol JC, Jimenez CR, Verheul HMW and Buffart TE: Proteomic analysis of miR-195 and miR-497 replacement reveals potential candidates that increase sensitivity to oxaliplatin in MSI/P53wt colorectal cancer cells. Cells. 8(1111)2019.PubMed/NCBI View Article : Google Scholar
21	Pang H, Xu X, Dai L, Wang K and Yao X: MicroRNA-195 is associated with regulating the pathophysiologic process of human laryngeal squamous cell carcinoma. Mol Med Rep. 17:5283–5291. 2018.PubMed/NCBI View Article : Google Scholar
22	Cheng JZ, Chen JJ, Wang ZG, Yu D and Zu YZ: The functional role of microRNAs in laryngeal carcinoma. Open Life Sci. 12:460–464. 2017.
23	Wang PP, Ding SY, Sun YY, Li YH and Fu WN: MYCT1 inhibits the adhesion and migration of laryngeal cancer cells potentially through repressing collagen VI. Front Oncol. 10(564733)2021.PubMed/NCBI View Article : Google Scholar
24	Song Y, Yang M, Zhang H, Sun Y, Tao Y, Li H, Zhang J, Li Y and Yang J: IL-17 affects the progression, metastasis, and recurrence of laryngeal cancer via the inhibition of apoptosis through activation of the PI3K/AKT/FAS/F pathways. J Immunol Res. 2020(2953191)2020.PubMed/NCBI View Article : Google Scholar
25	Li Y, Li D, Wang P, Zhu W and Yin W: Tetrandrine partially reverses multidrug resistance of human laryngeal cancer cells. J Int Med Res. 48(300060520944706)2020.PubMed/NCBI View Article : Google Scholar
26	Shang Y, Wang LQ, Guo QY and Shi TL: MicroRNA-196a overexpression promotes cell proliferation and inhibits cell apoptosis through PTEN/Akt/FOXO1 pathway. Int J Clin Exp Pathol. 8:2461–2472. 2015.PubMed/NCBI
27	Jin Y, Wang M, Hu H, Huang Q, Chen Y and Wang G: Overcoming stemness and chemoresistance in colorectal cancer through miR-195-5p-modulated inhibition of notch signaling. Int J Biol Macromol. 117:445–453. 2018.PubMed/NCBI View Article : Google Scholar
28	Li Y, Di C, Li W, Cai W, Tan X, Xu L, Yang L, Lou G and Yan Y: Oncomirs miRNA-221/222 and tumor suppressors miRNA-199a/195 are crucial miRNAs in liver cancer: A systematic analysis. Dig Dis Sci. 61:2315–2327. 2016.PubMed/NCBI View Article : Google Scholar
29	Yu X, Zhang Y, Ma X and Pertsemlidis A: miR-195 potentiates the efficacy of microtubule-targeting agents in non-small cell lung cancer. Cancer Lett. 427:85–93. 2018.PubMed/NCBI View Article : Google Scholar
30	Zheng J, Xu T, Chen F and Zhang Y: MiRNA-195-5p functions as a tumor suppressor and a predictive of poor prognosis in non-small cell lung cancer by directly targeting CIAPIN1. Pathol Oncol Res. 25:1181–1190. 2019.PubMed/NCBI View Article : Google Scholar
31	Kalluri R and Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest. 112:1776–1784. 2003.PubMed/NCBI View Article : Google Scholar
32	Kalluri R and Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 119:1420–1428. 2009.PubMed/NCBI View Article : Google Scholar
33	Wang Q, Wang F, Zhong W, Ling H, Wang J, Cui J, Xie T, Wen S and Chen J: RNA-binding protein RBM6 as a tumor suppressor gene represses the growth and progression in laryngocarcinoma. Gene. 697:26–34. 2019.PubMed/NCBI View Article : Google Scholar
34	Hwang KE, Kim HJ, Song IS, Park C, Jung JW, Park DS, Oh SH, Kim YS and Kim HR: Salinomycin suppresses TGF-β1-induced EMT by down-regulating MMP-2 and MMP-9 via the AMPK/SIRT1 pathway in non-small cell lung cancer. Int J Med Sci. 18:715–726. 2021.PubMed/NCBI View Article : Google Scholar
35	Karmakar D, Maity J, Mondal P, Shyam Chowdhury P, Sikdar N, Karmakar P, Das C and Sengupta S: E2F5 promotes prostate cancer cell migration and invasion through regulation of TFPI2, MMP-2 and MMP-9. Carcinogenesis. 41:1767–1780. 2020.PubMed/NCBI View Article : Google Scholar
36	Miles WO, Tschöp K, Herr A, Ji JY and Dyson NJ: Pumilio facilitates miRNA regulation of the E2F3 oncogene. Genes Dev. 26:356–368. 2012.PubMed/NCBI View Article : Google Scholar
37	Tang W, Tang J, Qin J, Geng Q, Zhou Z, Li B, Zhang J, Chen H, Xia Y and Wang X: Involvement of down-regulated E2F3 in Hirschsprung's disease. J Pediatr Surg. 48:813–817. 2013.PubMed/NCBI View Article : Google Scholar
38	Huang L, Luo J, Cai Q, Pan Q, Zeng H, Guo Z, Dong W, Huang J and Lin T: MicroRNA-125b suppresses the development of bladder cancer by targeting E2F3. Int J Cancer. 128:1758–1769. 2011.PubMed/NCBI View Article : Google Scholar
39	Ren XS, Yin MH, Zhang X, Wang Z, Feng SP, Wang GX, Luo YJ, Liang PZ, Yang XQ, He JX and Zhang BL: Tumor-suppressive microRNA-449a induces growth arrest and senescence by targeting E2F3 in human lung cancer cells. Cancer Lett. 344:195–203. 2014.PubMed/NCBI View Article : Google Scholar