MicroRNA‑150 suppresses p27<sup>Kip1</sup> expression and promotes cell proliferation in HeLa human cervical cancer cells

Oboshi,Wataru; Hayashi,Keisuke; Takeuchi,Hiroaki; Ikeda,Katsuhide; Yamaguchi,Yoshitaka; Kimura,Asako; Nakamura,Takehiro; Yukimasa,Nobuyasu

doi:10.3892/ol.2020.12073

MicroRNA‑150 suppresses p27^Kip1 expression and promotes cell proliferation in HeLa human cervical cancer cells

Authors:
- Wataru Oboshi
- Keisuke Hayashi
- Hiroaki Takeuchi
- Katsuhide Ikeda
- Yoshitaka Yamaguchi
- Asako Kimura
- Takehiro Nakamura
- Nobuyasu Yukimasa
View Affiliations

Affiliations: Department of Medical Technology and Sciences, International University of Health and Welfare, Narita Chiba 286‑8686, Japan, Department of Clinical Laboratory, Shikoku Central Hospital, Shikokuchuo, Ehime 799‑0193, Japan, Department of Medical Technology, Kagawa Prefectural University of Health Sciences, Takamatsu, Kagawa 761‑0123, Japan
Published online on: September 8, 2020 https://doi.org/10.3892/ol.2020.12073
Article Number: 210
Copyright: © Oboshi 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 (miRNAs) exert critical roles in the majority of biological and pathological processes. Recent studies have associated miR‑150 with a number of different cancer types. However, little is known about miR‑150 targets in cervical cancer. In the present study, the HeLa human cervical cancer cell line was transfected with hsa‑miR‑150‑5p mimics, hsa‑miR‑150‑5p inhibitors or miRNA controls. miR‑150 was predicted to bind the 3'untranslated region (3'UTR) of the CDKN1B gene, which encodes the cyclin‑dependent kinase inhibitor 1B (p27^Kip1). The direct binding between miR‑150 and the 3'UTR of CDKN1B was confirmed using dual‑luciferase reporter assays. The effects of miR‑150 on CDKN1B mRNA expression, p27^Kip1 protein expression, cell cycle and cell proliferation were determined using reverse‑transcription quantitative PCR, western blot analysis, flow cytometry and WST‑8 assays, respectively. miR‑150 was demonstrated to directly target the 3'UTR of CDKN1B in transfected HeLa cells. The expression of CDKN1B mRNA and p27^Kip1 protein was reduced by miR‑150 mimics, and increased by miR‑150 inhibitors. Moreover, the overexpression of miR‑150 promoted cell cycle progression from the G0/G1 to the S phase and led to a significant increase in HeLa cell proliferation. The results of the present study indicated that miR‑150 promotes HeLa cell cycle progression and proliferation via the suppression of p27^Kip1 expression.

View Figures

View References

1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 66:7–30. 2016. View Article : Google Scholar : PubMed/NCBI
3	Hagemann T, Bozanovic T, Hooper S, Ljubic A, Slettenaar VI, Wilson JL, Singh N, Gayther SA, Shepherd JH and Van Trappen PO: Molecular profiling of cervical cancer progression. Br J Cancer. 96:321–328. 2007. View Article : Google Scholar : PubMed/NCBI
4	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
5	Lai EC: miRNAs: Whys and wherefores of miRNA-mediated regulation. Curr Biol. 15:R458–R460. 2005. View Article : Google Scholar : PubMed/NCBI
6	Friedman RC, Farh KK, Burge CB and Bartel DP: Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19:92–105. 2009. View Article : Google Scholar : PubMed/NCBI
7	Ebert MS and Sharp PA: Roles for microRNAs in conferring robustness to biological processes. Cell. 149:515–524. 2012. View Article : Google Scholar : PubMed/NCBI
8	Calin GA and Croce CM: MicroRNA signatures in human cancers. Nat Rev Cancer. 6:857–866. 2006. View Article : Google Scholar : PubMed/NCBI
9	Farazi TA, Spitzer JI, Morozov P and Tuschl T: miRNAs in human cancer. J Pathol. 223:102–115. 2011. View Article : Google Scholar : PubMed/NCBI
10	Wang F, Ren X and Zhang X: Role of microRNA-150 in solid tumors. Oncol Lett. 10:11–16. 2015. View Article : Google Scholar : PubMed/NCBI
11	Feng J, Yang Y, Zhang P, Wang F, Ma Y, Qin H and Wang Y: miR-150 functions as a tumour suppressor in human colorectal cancer by targeting c-Myb. J Cell Mol Med. 18:2125–2134. 2014. View Article : Google Scholar : PubMed/NCBI
12	Srivastava SK, Bhardwaj A, Singh S, Arora S, Wang B, Grizzle WE and Singh AP: MicroRNA-150 directly targets MUC4 and suppresses growth and malignant behavior of pancreatic cancer cells. Carcinogenesis. 32:1832–1839. 2011. View Article : Google Scholar : PubMed/NCBI
13	Huang S, Chen Y, Wu W, Ouyang N, Chen J, Li H, Liu X, Su F, Lin L and Yao Y: miR-150 promotes human breast cancer growth and malignant behavior by targeting the pro-apoptotic purinergic P2X7 receptor. PLoS One. 8:e807072013. View Article : Google Scholar : PubMed/NCBI
14	Jin M, Yang Z, Ye W, Xu H and Hua X: MicroRNA-150 predicts a favorable prognosis in patients with epithelial ovarian cancer, and inhibits cell invasion and metastasis by suppressing transcriptional repressor ZEB1. PLoS One. 9:e1039652014. View Article : Google Scholar : PubMed/NCBI
15	Dezhong L, Xiaoyi Z, Xianlian L, Hongyan Z, Guohua Z, Bo S, Shenglei Z and Lian Z: miR-150 is a factor of survival in prostate cancer patients. J BUON. 20:173–179. 2015.PubMed/NCBI
16	Zhang J, Luo N, Luo Y, Peng Z, Zhang T and Li S: MicroRNA-150 inhibits human CD133-positive liver cancer stem cells through negative regulation of the transcription factor c-Myb. Int J Oncol. 40:747–756. 2012.PubMed/NCBI
17	Katada T, Ishiguro H, Kuwabara Y, Kimura M, Mitui A, Mori Y, Ogawa R, Harata K and Fujii Y: MicroRNA expression profile in undifferentiated gastric cancer. Int J Oncol. 34:537–542. 2009.PubMed/NCBI
18	Li J, Hu L, Tian C, Lu F, Wu J and Liu L: MicroRNA-150 promotes cervical cancer cell growth and survival by targeting FOXO4. BMC Mol Biol. 16:242015. View Article : Google Scholar : PubMed/NCBI
19	Zhou L, Qi X, Potashkin JA, Abdul-Karim FW and Gorodeski GI: MicroRNAs miR-186 and miR-150 down-regulate expression of the pro-apoptotic purinergic P2X7 receptor by activation of instability sites at the 3′-untranslated region of the gene that decrease steady-state levels of the transcript. J Biol Chem. 283:28274–2886. 2008. View Article : Google Scholar : PubMed/NCBI
20	Zhu J and Han S: miR-150-5p promotes the proliferation and epithelial-mesenchymal transition of cervical carcinoma cells via targeting SRCIN1. Pathol Res Pract. 215:738–747. 2019. View Article : Google Scholar : PubMed/NCBI
21	Boura E, Silhan J, Herman P, Vecer J, Sulc M, Teisinger J, Obsilova V and Obsil T: Both the N-terminal loop and wing W2 of the forkhead domain of transcription factor Foxo4 are important for DNA binding. J Biol Chem. 282:8265–8275. 2007. View Article : Google Scholar : PubMed/NCBI
22	Wang Q, Li X, Wang L, Feng YH, Zeng R and Gorodeski G: Antiapoptotic effects of estrogen in normal and cancer human cervical epithelial cells. Endocrinology. 145:5568–5579. 2004. View Article : Google Scholar : PubMed/NCBI
23	Cabodi S, del Pilar Camacho-Leal M, Di Stefano P and Defilippi P: Integrin signalling adaptors: Not only figurants in the cancer story. Nat Rev Cancer. 10:858–870. 2010. View Article : Google Scholar : PubMed/NCBI
24	Zhang Z, Wang J, Li J, Wang X and Song W: MicroRNA-150 promotes cell proliferation, migration, and invasion of cervical cancer through targeting PDCD4. Biomed Pharmacother. 97:511–517. 2018. View Article : Google Scholar : PubMed/NCBI
25	Polyak K, Lee MH, Erdjument-Bromage H, Koff A, Roberts JM, Tempst P and Massagué J: Cloning of p27Kip1, a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals. Cell. 78:59–66. 1994. View Article : Google Scholar : PubMed/NCBI
26	Toyoshima H and Hunter T: p27, a novel inhibitor of G1 cyclin-Cdk protein kinase activity, is related to p21. Cell. 78:67–74. 1994. View Article : Google Scholar : PubMed/NCBI
27	Pizzini S, Bisognin A, Mandruzzato S, Biasiolo M, Facciolli A, Perilli L, Rossi E, Esposito G, Rugge M, Pilati P, et al: Impact of microRNAs on regulatory networks and pathways in human colorectal carcinogenesis and development of metastasis. BMC Genomics. 14:5892013. View Article : Google Scholar : PubMed/NCBI
28	Ma Y, Zhang P, Wang F, Zhang H, Yang J, Peng J, Liu W and Qin H: miR-150 as a potential biomarker associated with prognosis and therapeutic outcome in colorectal cancer. Gut. 61:1447–1453. 2012. View Article : Google Scholar : PubMed/NCBI
29	Sun W, Zhang Z, Wang J, Shang R, Zhou L, Wang X, Duan J, Ruan B, Gao Y, Dai B, et al: MicroRNA-150 suppresses cell proliferation and metastasis in hepatocellular carcinoma by inhibiting the GAB1-ERK axis. Oncotarget. 7:11595–11608. 2016. View Article : Google Scholar : PubMed/NCBI
30	Yokobori T, Suzuki S, Tanaka N, Inose T, Sohda M, Sano A, Sakai M, Nakajima M, Miyazaki T, Kato H and Kuwano H: miR-150 is associated with poor prognosis in esophageal squamous cell carcinoma via targeting the EMT inducer ZEB1. Cancer Sci. 104:48–54. 2013. View Article : Google Scholar : PubMed/NCBI
31	Li H, Ouyang R, Wang Z, Zhou W, Chen H, Jiang Y, Zhang Y, Li H, Liao M, Wang W, et al: miR-150 promotes cellular metastasis in non-small cell lung cancer by targeting FOXO4. Sci Rep. 6:390012016. View Article : Google Scholar : PubMed/NCBI
32	Yin QW, Sun XF, Yang GT, Li XB, Wu MS and Zhao J: Increased expression of microRNA-150 is associated with poor prognosis in non-small cell lung cancer. Int J Clin Exp Pathol. 8:842–846. 2015.PubMed/NCBI
33	Zhang N, Wei X and Xu L: miR-150 promotes the proliferation of lung cancer cells by targeting P53. FEBS Lett. 587:2346–2351. 2013. View Article : Google Scholar : PubMed/NCBI
34	Wu Q, Jin H, Yang Z, Luo G, Lu Y, Li K, Ren G, Su T, Pan Y, Feng B, et al: miR-150 promotes gastric cancer proliferation by negatively regulating the pro-apoptotic gene EGR2. Biochem Biophys Res Commun. 392:340–345. 2010. View Article : Google Scholar : PubMed/NCBI
35	Filipowicz W, Bhattacharyya SN and Sonenberg N: Mechanisms of post-transcriptional regulation by microRNAs: Are the answers in sight? Nat Rev Genet. 9:102–114. 2008. View Article : Google Scholar : PubMed/NCBI