MicroRNA‑22 enhances radiosensitivity in cervical cancer cell lines via direct inhibition of c‑Myc binding protein, and the subsequent reduction in hTERT expression

Nakamura,Mayumi; Hayashi,Masami; Konishi,Hiromi; Nunode,Misa; Ashihara,Keisuke; Sasaki,Hiroshi; Terai,Yoshito; Ohmichi,Masahide

doi:10.3892/ol.2020.11344

MicroRNA‑22 enhances radiosensitivity in cervical cancer cell lines via direct inhibition of c‑Myc binding protein, and the subsequent reduction in hTERT expression

Authors:
- Mayumi Nakamura
- Masami Hayashi
- Hiromi Konishi
- Misa Nunode
- Keisuke Ashihara
- Hiroshi Sasaki
- Yoshito Terai
- Masahide Ohmichi
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, Osaka Medical College, Takatsuki, Osaka 569‑8686, Japan, Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine, Kobe, Hyogo 650‑0017, Japan
Published online on: January 23, 2020 https://doi.org/10.3892/ol.2020.11344
Pages: 2213-2222
Copyright: © Nakamura 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

MicroRNAs (miRs) influence the expression of their target genes post‑transcriptionally and serve an important role in multiple cellular processes. The downregulation of miR‑22 is associated with a poor prognosis in cervical cancer. However, the mechanisms underlying miR‑22‑mediated gene regulation and its function are yet to be elucidated. In the present study, the effect of miR‑22 expression on the radiosensitivity of cervical cancer was investigated. First, miR‑22 was either up‑ or downregulated to evaluate the regulation of the MYC‑binding protein (MYCBP) in four cervical cancer cell lines (C‑4I, SKG‑II and SiHa). Notably, MYCBP expression was inversely associated with miR‑22 induction. A dual‑luciferase reporter gene assay revealed that miR‑22 directly targets the MYCBP 3'‑untranslated region. Subsequently, the level of human telomerase reverse transcriptase component (hTERT; an E‑box‑containing c‑Myc target gene) was analyzed after the up‑ or downregulation of miR‑22. Notably, miR‑22‑mediated repression of MYCBP reduced hTERT expression. In addition, the influence of miR‑22 on radiosensitivity in C‑4I, SKG‑II and SiHa cells was examined using a clonogenic assay and in mouse xenograft models. Upregulation of miR‑22 was associated with increased radiosensitivity. Furthermore, lentiviral transduction of miR‑22 reduced the Ki‑67 index while increasing the TUNEL index in xenograft tissue. The current findings indicate the potential utility of miR‑22 in radiotherapy for cervical cancer.

View Figures

View References

1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 coutries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Yagi A, Ueda Y, Kakuda M, Tanaka Y, Ikeda S, Matsuzaki S, Kobayashi E, Morishima T, Miyashiro I, Fukui K, et al: Epidemiologic and clinical analysis of cervical cancer using data from the population-based Osaka cancer regstry. Cancer Res. 79:1252–1259. 2019. View Article : Google Scholar : PubMed/NCBI
3	Fokom Domgue J and Schmeler KM: Conservative management of cervical cancer: Current status and obstetrical implications. Best Pract Res Clin Obstet Gynaecol. 55:79–92. 2019. View Article : Google Scholar : PubMed/NCBI
4	Chohen PA, Jhingran A, Oaknin A and Denny L: Cercvical cancer. Lancet. 393:169–182. 2019. View Article : Google Scholar : PubMed/NCBI
5	Wright JD, Matsuo K, Huang Y, Tergas AI, Hou JY, Khoury-Collado F, St Clair CM, Ananth CV, Neugut AI and Hershman DL: Prognostic performance of the 2018 international federation of gynecololgy and obstetrics cervical cancer staging guidelines. Obstet Gynecol. 134:49–57. 2019. View Article : Google Scholar : PubMed/NCBI
6	Pal J, Gold JS, Munshi NC and Shammas MA: Biology of telomeres: Importance in etiology of esophageal cancer and as therapeutic target. Transl Res. 162:364–370. 2013. View Article : Google Scholar : PubMed/NCBI
7	Lord RV, Salonga D, Danenberg KD, Peters JH, DeMeester TR, Park JM, Johansson J, Skinner KA, Chandrasoma P, DeMeester SR, et al: Telomerase reverse transcriptase expression is increased early in the Barrett's metaplasia, dysplasia, adenocarcinoma sequence. J Gastrointest Surg. 4:135–142. 2000. View Article : Google Scholar : PubMed/NCBI
8	Hoang-Vu C, Boltze C, Gimm O, Poremba C, Dockhorn-Dworniczak B, Köhrle J, Rath FW and Dralle H: Expression of telomerase genes in thyroid carcinoma. Int J Oncol. 21:265–272. 2002.PubMed/NCBI
9	Wang R, Lin F, Wang X, Gao P, Dong K, Wei SH, Cheng SY and Zhang HZ: The therapeutic potential of survivin promoter-driven siRNA on suppressing tumor growth and enhancing radiosensitivity of human cervical carcinoma cells via downregulating hTERT gene expression. Cancer Biol Ther. 6:1295–1301. 2007. View Article : Google Scholar : PubMed/NCBI
10	Kyo S, Takakura M, Taira T, Kanaya T, Itoh H, Yutsudo M, Ariga H and Inoue M: Sp1 cooperates with c-Myc to activate transcription of the human telomerase reverse transcriptase gene (hTERT). Nucleic Acids Res. 28:669–677. 2000. View Article : Google Scholar : PubMed/NCBI
11	Ambros V: MicroRNAs: Tiny regulators with great potential. Cell. 107:823–826. 2001. View Article : Google Scholar : PubMed/NCBI
12	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
13	Kaur A, Mackin ST, Schlosser K, Wong FL, Elharram M, Delles C, Stewart DJ, Dayan N, Landry T and Pilote L: Systemativ review of microRNA biomarkers in acute coronary syndrome and stable coronary artery disease. Cardiovasc Res. (pii): cvz3022019.(Epub ahead of print). View Article : Google Scholar : PubMed/NCBI
14	Ji C and Guo X: The clinical potential of circulating microRNAs in obesity. Nat Rev Endocrinol. 15:731–743. 2019. View Article : Google Scholar : PubMed/NCBI
15	Zhang L, Chen B and Ding D: Decreased microRNA-22 is associated with poor prognosis in cervical cancer. Int J Clin Exp Pathol. 10:9515–9520. 2017.PubMed/NCBI
16	Xiong J, Du Q and Liang Z: Tumor-suppressive microRNA-22 inhibits the transcription of E-box-containing c-Myc target genes by silencing c-Myc binding protein. Oncogene. 29:4980–4988. 2010. View Article : Google Scholar : PubMed/NCBI
17	Grimson A, Farh KK, Johnston WK, Garrett-Engele P, Lim LP and Bartel DP: MicroRNA targeting specificity in mammals: Determinants beyond seed pairing. Mol Cell. 27:91–105. 2007. View Article : Google Scholar : PubMed/NCBI
18	Wong N and Wang X: MiRDB: An online resource for microRNA target prediction and functional annotations. Nucleic Acids Res. 43((Database Issue)): D146–D152. 2015. View Article : Google Scholar : PubMed/NCBI
19	Livak KJ and Schimittgen 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. View Article : Google Scholar : PubMed/NCBI
20	Ono YJ, Hayashi M, Tanabe A, Hayashi A, Kanemura M, Terai Y and Ohmichi M: Estradiol-mediated hepatocyte growth factor is involved in the implantation of endometriotic cells via the mesothelial-to-mesenchymal transition in the peritoneum. Am J Physiol Endocrinol Metab. 308:E950–E959. 2015. View Article : Google Scholar : PubMed/NCBI
21	Franken NA, Rodermond HM, Stap J, Haveman J and van Bree C: Clonogenic assay of cells in vitro. Nat Protoc. 1:2315–2319. 2006. View Article : Google Scholar : PubMed/NCBI
22	Taira T, Maeda J, Onishi T, Kitaura H, Yoshida S, Kato H, Ikeda M, Tamai K, Iguchi-Ariga SM and Ariga H: AMY-1, a novel C-MYC binding protein that stimulates transcription activity of C-MYC. Genes Cells. 3:549–565. 1998. View Article : Google Scholar : PubMed/NCBI
23	Wu KJ, Grandori C, Amacker M, Simon-Vermot N, Polack A, Lingner J and Dalla-Favera R: Direct activation of TERT transcription by c-MYC. Nat Genet. 21:220–224. 1999. View Article : Google Scholar : PubMed/NCBI
24	Zhang W and Xing L: RNAi gene therapy of SiHa cells via targeting human TERT induces growth inhibition and enhances radiosensitivity. Int J Oncol. 43:1228–1234. 2013. View Article : Google Scholar : PubMed/NCBI
25	Nakatani F, Ferracin M, Manara MC, Ventura S, Del Monaco V, Ferrari S, Alberghini M, Grilli A, Knuutila S, Schaefer KL, et al: MiR-34a predicts survival of Ewing's sarcoma patients and directly influences cell chemo-sensitivity and malignancy. J Pathol. 226:796–805. 2012. View Article : Google Scholar : PubMed/NCBI
26	Lagos-Quintana M, Rauhut R, Lendeckel W and Tuschl T: Identification of novel genes coding for small expressed RNAs. Science. 294:853–858. 2001. View Article : Google Scholar : PubMed/NCBI
27	Xin M, Qiao Z, Li J, Liu J, Song S, Zhao X, Miao P, Tang T, Wang L, Liu W, et al: MiR-22 inhibits tumor growth and metastasis by targeting ATP citrate lyase: Evidence in osteosarcoma, prostate cancer, cervical cancer and lung cancer. Oncotarget. 7:44252–44265. 2016. View Article : Google Scholar : PubMed/NCBI
28	Li J, Liang S, Yu H, Zhang J, Ma D and Lu X: An inhibitory effect of miR-22 on cell migration and invasion in ovarian cancer. Gynecol Oncol. 119:543–538. 2010. View Article : Google Scholar : PubMed/NCBI
29	Zuo QF, Cao LY, Yu T, Gong L, Wang LN, Zhao YL, Xiao B and Zou QM: MicroRNA-22 inhibits tumor growth and metastasis in gastric cancer by directly targeting MMP14 and Snail. Cell Death Dis. 6:e20002015. View Article : Google Scholar : PubMed/NCBI
30	Zhang H, Tang J, Li C, Kong J, Wang J, Wu Y, Xu E and Lai M: MiR-22 regulates 5-FU sensitivity by inhibiting autophagy and promoting apoptosis in colorectal cancer cells. Cancer Lett. 356:781–790. 2015. View Article : Google Scholar : PubMed/NCBI
31	Zhang T, Xue X and Peng H: Therapeutic delivery of miR-29b enhances radiosensitivity in cervical cancer. Mol Ther. 27:1183–1194. 2019. View Article : Google Scholar : PubMed/NCBI
32	Pedroza-Torres A, Campos-Parra AD, Millan-Catalan O, Loissell-Baltazar YA, Zamudio-Meza H, Cantú de León D, Montalvo-Esquivel G, Isla-Ortiz D, Herrera LA, Ángeles-Zaragoza Ó, et al: MicroRNA-125 modulates radioresistance through targeting p21 in cervical cancer. Oncol Rep. 39:1532–1540. 2018.PubMed/NCBI
33	Zhang X, Li Y, Wang D and Wei X: MiR-22 suppresses tumorigenesis and improves radiosensitivity of breast cancer cells by targeting Sirt1. Biol Res. 50:272017. View Article : Google Scholar : PubMed/NCBI
34	Liu Z, Li T, Deng S, Fu S, Zhou X and He Y: Radiation induces apoptosis and osteogenic impairment through miR-22-mediated intracellular oxidative stress in bone marrow mesenchymal stem cells. Stem Cells Int. 2018:58454022018. View Article : Google Scholar : PubMed/NCBI
35	Sakamuro D and Prendergast GC: New Myc-interacting proteins: A second Myc network emerges. Oncogene. 18:2942–2954. 1999. View Article : Google Scholar : PubMed/NCBI
36	Wang H, Yan X, Ji LY, Ji XT, Wang P, Guo SW and Li SZ: MiR-139 functions as an antioncomir to repress glioma progression through targeting IGF-1 R, AMY-1, and PGC-1β. Technol Cancer Res Treat. 16:497–511. 2017. View Article : Google Scholar : PubMed/NCBI
37	Gong L, Xia Y, Qian Z, Shi J, Luo J, Song G, Xu J and Ye Z: Overexpression of MYC binding protein promotes invasion and migration in gastric cancer. Oncol Lett. 15:5243–5249. 2018.PubMed/NCBI