MicroRNA‑145 sensitizes cervical cancer cells to low‑dose irradiation by downregulating OCT4 expression

Yan,Siqi; Li,Xiangjun; Jin,Qiao; Yuan,Jun

doi:10.3892/etm.2016.3731

MicroRNA‑145 sensitizes cervical cancer cells to low‑dose irradiation by downregulating OCT4 expression

Authors:
- Siqi Yan
- Xiangjun Li
- Qiao Jin
- Jun Yuan
View Affiliations

Affiliations: Department of Oncology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R. China, Department of Oncology, The Second People's Hospital, Changsha, Hunan 410000, P.R. China, Department of Oncology, Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China
Published online on: September 20, 2016 https://doi.org/10.3892/etm.2016.3731
Pages: 3130-3136

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

Poor elucidation of the mechanisms involved in regulating the radiosensitivity of cancers prevents the extensive application of low‑dose radiotherapy in clinical settings. The present study was conducted to investigate the role of microRNA‑145 (miR‑145) in the modulation of cervical cancer cell radiosensitivity, as well as to identify the underlying target of miR‑145 during this process. Cervical cancer tera cells were initially exposed to doses of radiation between 1 and 6 Gy before the assessments of the cell viability and apoptosis rate. Irradiation at dose of 1 Gy was screened as optimum dose and used in subsequent experiments. A dual luciferase reporter assay was performed to demonstrate that octamer‑binding transcription factor 4 (OCT4) is a target of miR‑145 in cervical cancer. Consequently, OCT4 was suggested to be a target of miR‑145, as a dual luciferase vector that was ligated to a fragment corresponding to the predicted target site of miR‑145 in OCT4 3'‑UTR showed an 83% reduction in fluorescence. Following exposure to 1 Gy irradiation, tera cells transfected with miR‑145 mimics, which showed downregulation of OCT4 and cyclin D1, had lower cell viability and cell migration rate and higher apoptosis rate compared to non‑transfected cells. However, the co‑transfection of miR‑145 mimics and OCT4 expression vector restored OCT4 and cyclin D1 expression levels and made no significant difference in terms of cell viability, cell migration rate and apoptosis rate. The present results indicate that miR‑145 increases the radiosensitivity of cervical cancer cells by silencing OCT4, that cyclin D1 is putatively under the positive regulation of OCT4 and mediates miR‑145 function.

View Figures

View References

1	Wang X, Tang S, Le SY, Lu R, Rader JS, Meyers C and Zheng ZM: Aberrant expression of oncogenic and tumor-suppressive microRNAs in cervical cancer is required for cancer cell growth. PLoS One. 3:25572008. View Article : Google Scholar
2	Mongula J, Slangen B, Lambregts D, Bakers F, Mahesh S, Lutgens L, Van Gorp T, Vliegen R, Kruitwagen R and Beets-Tan R: Predictive criteria for MRI-based evaluation of response both during and after radiotherapy for cervical cancer. J Contemp Brachytherapy. 8:181–188. 2016. View Article : Google Scholar : PubMed/NCBI
3	Kitahara O, Katagiri T, Tsunoda T, Harima Y and Nakamura Y: Classification of sensitivity or resistance of cervical cancers to ionizing radiation according to expression profiles of 62 genes selected by cDNA microarray analysis. Neoplasia. 4:295–303. 2002. View Article : Google Scholar : PubMed/NCBI
4	Liu SS, Chan KY, Leung RC, Law HK, Leung TW and Ngan HY: Enhancement of the radiosensitivity of cervical cancer cells by overexpressing p73alpha. Mol Cancer Ther. 5:1209–1215. 2006. View Article : Google Scholar : PubMed/NCBI
5	Liu R, Wang X, Tian JH, Yang K, Wang J, Jiang L and Hao XY: High dose rate versus low dose rate intracavity brachytherapy for locally advanced uterine cervix cancer. Cochrane Database Syst Rev. 9:CD0075632014.
6	Hareyama M, Sakata K, Oouchi A, Nagakura H, Shido M, Someya M and Koito K: High-dose-rate versus low-dose-rate intracavitary therapy for carcinoma of the uterine cervix: A randomized trial. Cancer. 94:117–124. 2002. View Article : Google Scholar : PubMed/NCBI
7	Cui H, Qin Q, Yang M, Zhang H, Liu Z, Yang Y, Chen X, Zhu H, Wang D, Meng C, et al: Bortezomib enhances the radiosensitivity of hypoxic cervical cancer cells by inhibiting HIF-1α expression. Int J Clin Exp Pathol. 8:9032–9041. 2015.PubMed/NCBI
8	Reshmi G and Pillai MR: Beyond HPV: Oncomirs as new players in cervical cancer. FEBS Lett. 582:4113–4116. 2008. View Article : Google Scholar : PubMed/NCBI
9	Xue M, Zhao L, Yang F, Li Z and Li G: MicroRNA-145 inhibits the malignant phenotypes of gastric carcinoma cells via downregulation of fascin 1 expression. Mol Med Rep. 13:1033–1039. 2016.PubMed/NCBI
10	Han T, Yi XP, Liu B, Ke MJ and Li YX: MicroRNA-145 suppresses cell proliferation, invasion and migration in pancreatic cancer cells by targeting NEDD9. Mol Med Rep. 11:4115–4120. 2015.PubMed/NCBI
11	Wang S, Bian C, Yang Z, Bo Y, Li J, Zeng L, Zhou H and Zhao RC: miR-145 inhibits breast cancer cell growth through RTKN. Int J Oncol. 34:1461–1466. 2009.PubMed/NCBI
12	Ostenfeld MS, Bramsen JB, Lamy P, Villadsen SB, Fristrup N, Sørensen KD, Ulhøi B, Borre M, Kjems J, Dyrskjøt L and Orntoft TF: miR-145 induces caspase-dependent and -independent cell death in urothelial cancer cell lines with targeting of an expression signature present in Ta bladder tumors. Oncogene. 29:1073–1084. 2010. View Article : Google Scholar : PubMed/NCBI
13	Gu TT, Liu SY and Zheng PS: Cytoplasmic NANOG-positive stromal cells promote human cervical cancer progression. Am J Pathol. 181:652–661. 2012. View Article : Google Scholar : PubMed/NCBI
14	Vaiphei K, Sinha SK and Kochhar R: Comparative analysis of Oct4 in different histological subtypes of esophageal squamous cell carcinomas in different clinical conditions. Asian Pac J Cancer Prev. 15:3519–3524. 2014. View Article : Google Scholar : PubMed/NCBI
15	Vargas TH, Pulz LH, Barra CN, Kleeb SR, Xavier JG, Catão-Dias JL, Fukumasu H, Nishiya AT and Strefezzi RF: Immunohistochemical expression of the pluripotency factor OCT4 in canine mast cell tumours. J Comp Pathol. 153:251–255. 2015. View Article : Google Scholar : PubMed/NCBI
16	Tsai LL, Yu CC, Chang YC, Yu CH and Chou MY: Markedly increased Oct4 and Nanog expression correlates with cisplatin resistance in oral squamous cell carcinoma. J Oral Pathol Med. 40:621–628. 2011. View Article : Google Scholar : PubMed/NCBI
17	Lo WL, Chien Y, Chiou GY, Tseng LM, Hsu HS, Chang YL, Lu KH, Chien CS, Wang ML, Chen YW, et al: Nuclear localization signal-enhanced RNA interference of EZH2 and Oct4 in the eradication of head and neck squamous cell carcinoma-derived cancer stem cells. Biomaterials. 33:3693–3709. 2012. View Article : Google Scholar : PubMed/NCBI
18	Kuo KK, Lee KT, Chen KK, Yang YH, Lin YC, Tsai MH, Wuputra K, Lee YL, Ku CC, Miyoshi H, et al: Positive feedback loop of OCT4 and c-JUN expedites cancer stemness in liver cancer. Stem Cells. Jun 24–2016.(Epub ahead of print). View Article : Google Scholar
19	Lu CS, Shieh GS, Wang CT, Su BH, Su YC, Chen YC, Su WC, Wu P, Yang WH, Shiau AL and Wu CL: Chemotherapeutics-induced Oct4 expression contributes to drug resistance and tumor recurrence in bladder cancer. Oncotarget. May 26–2016.(Epub ahead of print).
20	Xu N, Papagiannakopoulos T, Pan G, Thomson JA and Kosik KS: MicroRNA-145 regulates OCT4, SOX2 and KLF4 and represses pluripotency in human embryonic stem cells. Cell. 137:647–658. 2009. View Article : Google Scholar : PubMed/NCBI
21	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
22	Dote H, Burgan WE, Camphausen K and Tofilon PJ: Inhibition of hsp90 compromises the DNA damage response to radiation. Cancer Res. 66:9211–9220. 2006. View Article : Google Scholar : PubMed/NCBI
23	Shuryak I and Brenner DJ: A model of interactions between radiation-induced oxidative stress, protein and DNA damage in Deinococcus radiodurans. J Theor Biol. 261:305–317. 2009. View Article : Google Scholar : PubMed/NCBI
24	Li C, Yan Y, Ji W, Bao L, Qian H, Chen L, Wu M, Chen H, Li Z and Su C: OCT4 positively regulates Survivin expression to promote cancer cell proliferation and leads to poor prognosis in esophageal squamous cell carcinoma. PLoS One. 7:e496932012. View Article : Google Scholar : PubMed/NCBI
25	Hu T, Liu S, Breiter DR, Wang F, Tang Y and Sun S: Octamer 4 small interfering RNA results in cancer stem cell-like cell apoptosis. Cancer Res. 68:6533–6540. 2008. View Article : Google Scholar : PubMed/NCBI
26	Lin Y, Yang Y, Li W, Chen Q, Li J, Pan X, Zhou L, Liu C and Chen C: Aself-renewal and survival of embryonal carcinoma cells. Mol Cell. 48:627–640. 2012. View Article : Google Scholar : PubMed/NCBI
27	Zhang Z, Zhu Y, Lai Y, Wu X, Feng Z, Yu Y, Bast RC Jr, Wan X, Xi X and Feng Y: Follicle-stimulating hormone inhibits apoptosis in ovarian cancer cells by regulating the OCT4 stem cell signaling pathway. Int J Oncol. 43:1194–1204. 2013.PubMed/NCBI
28	Wang YD, Cai N, Wu XL, Cao HZ, Xie LL and Zheng PS: OCT4 promotes tumorigenesis and inhibits apoptosis of cervical cancer cells by miR-125b/BAK1 pathway. Cell Death Dis. 4:e7602013. View Article : Google Scholar : PubMed/NCBI
29	Jeselsohn R, Brown NE, Arendt L, Klebba I, Hu MG, Kuperwasser C and Hinds PW: Cyclin D1 kinase activity is required for the self-renewal of mammary stem and progenitor cells that are targets of MMTV-ErbB2 tumorigenesis. Cancer Cell. 17:65–76. 2010. View Article : Google Scholar : PubMed/NCBI
30	Shimura T, Noma N, Oikawa T, Ochiai Y, Kakuda S, Kuwahara Y, Takai Y, Takahashi A and Fukumoto M: Activation of the AKT/Cyclin D1/Cdk4 survival signaling pathway in radioresistant cancer stem cells. Oncogenesis. 1:e122012. View Article : Google Scholar : PubMed/NCBI
31	Chu Q, Han N, Yuan X, Nie X, Wu H, Chen Y, Guo M, Yu S and Wu K: DACH1 inhibits Cyclin D1 expression, cellular proliferation and tumor growth of renal cancer cells. J Hematol Oncol. 7:732014. View Article : Google Scholar : PubMed/NCBI
32	Jin Q, Li X and Cao P: EphA2 modulates radiosensitive of hepatocellular carcinoma cells via p38/mitogen-activated protein kinase-mediated signal pathways. Kaohsiung J Med Sci. 31:510–517. 2015. View Article : Google Scholar : PubMed/NCBI
33	Suresh TN, Hemalatha A, Kumar ML Harendra and Mohiyuddin SM Azeem: Evaluation of histomorphological and immunohistochemical parameters as biomarkers of cervical lymph node metastasis in squamous cell carcinoma of oral cavity: A retrospective study. J Oral Maxillofac Pathol. 19:18–24. 2015. View Article : Google Scholar : PubMed/NCBI
34	Hwang SJ, Lee HW, Kim HR, Song HJ, Lee DH, Lee H, Shin CH, Joung JG, Kim DH, Joo KM and Kim HH: Overexpression of microRNA-95-3p suppresses brain metastasis of lung adenocarcinoma through downregulation of Cyclin D1. Oncotarget. 6:20434–20448. 2015. View Article : Google Scholar : PubMed/NCBI
35	Chen YJ, Lee LY, Chao YK, Chang JT, Lu YC, Li HF, Chiu CC, Li YC, Li YL, Chiou JF and Cheng AJ: DSG3 facilitates cancer cell growth and invasion through the DSG3-plakoglobin-TCF/LEF-Myc/cyclin D1/MMP signaling pathway. PLoS One. 8:e640882013. View Article : Google Scholar : PubMed/NCBI
36	Atkinson RL, Zhang M, Diagaradjane P, Peddibhotla S, Contreras A, Hilsenbeck SG, Woodward WA, Krishnan S, Chang JC and Rosen JM: Thermal enhancement with optically activated gold nanoshells sensitizes breast cancer stem cells to radiation therapy. Sci Transl Med. 2:55ra792010. View Article : Google Scholar : PubMed/NCBI
37	Card DA, Hebbar PB, Li L, Trotter KW, Komatsu Y, Mishina Y and Archer TK: Oct4/Sox2-regulated miR-302 targets Cyclin D1 in human embryonic stem cells. Mol Cell Biol. 28:6426–6438. 2008. View Article : Google Scholar : PubMed/NCBI