MicroRNA‑744‑5p inhibits glioblastoma malignancy by suppressing replication factor C subunit 2

Fan,Fei; Yao,Dongxiao; Yan,Pengfei; Jiang,Xiaobing; Hu,Jie

doi:10.3892/ol.2021.12869

MicroRNA‑744‑5p inhibits glioblastoma malignancy by suppressing replication factor C subunit 2

Authors:
- Fei Fan
- Dongxiao Yao
- Pengfei Yan
- Xiaobing Jiang
- Jie Hu
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Affiliations: Department of Neurosurgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China, Department of Neurosurgery, General Hospital of the Yangtze River Shipping, Jiangan, Wuhan, Hubei 430010, P.R. China
Published online on: June 15, 2021 https://doi.org/10.3892/ol.2021.12869
Article Number: 608
Copyright: © Fan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Glioblastoma (GBM) is the most common malignant primary brain tumor, accounting for ~57% of all gliomas and 48% of all malignant primary central nervous system tumors in the United States. Abnormal expression of the replication factor C subunit 2 (RFC2) gene and microRNA (miR)‑744‑5p is associated with tumorigenic characteristics, including cellular proliferation, migration and invasiveness. However, the mechanism underlying the interaction between miR‑744‑5p and RFC2 in GBM remains unknown. Reverse transcription‑quantitative (RT‑q) PCR analysis of RFC2 and miR‑744‑5p was performed using GBM tumor tissues and cells, and the association between miR‑744‑5p and RFC2 was determined by dual‑luciferase reporter assay. Cell Counting Kit 8, 5‑bromo‑2‑deoxyuridine (BrdU), wound‑healing and cellular adhesion assays, as well as the detection of caspase‑3 activity and western blotting were used to detect cellular proliferation, migration and adhesion, caspase‑3 activity, and Bax and Bcl‑2 protein expression, respectively, in GBM cells. The results of the present study demonstrated that RFC2 expression was increased in GBM tissues and cell lines. Overexpression of RFC2 promoted cellular proliferation, migration, adhesion and an increase in Bcl‑2 protein levels, and suppressed cellular caspase‑3 activity and Bax protein expression, while silencing RFC2 resulted in the opposite effect. The effects of miR‑744‑5p inhibition were similar to those of RFC2 overexpression. Moreover, miR‑744‑5p was found to target RFC2 in GBM cells, and inhibiting the expression of RFC2 suppressed GBM tumorigenesis. In conclusion, the present study demonstrated that miR‑744‑5p targets RFC2 and suppresses the progression of GBM by repressing cellular proliferation, migration and Bcl‑2 protein expression, and effectively promoting caspase‑3 activity and Bax protein expression. These findings suggest a new target for the clinical treatment and improved prognosis of patients with GBM in the future.

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1	Wen PY and Kesari S: Malignant gliomas in adults. New Engl J Med. 359:492–507. 2008. View Article : Google Scholar : PubMed/NCBI
2	Ostrom QT, Gittleman H, Truitt G, Boscia A, Kruchko C and Barnholtz-Sloan JS: CBTRUS statistical report: Primary brain and other central nervous system tumors diagnosed in the United States in 2011–2015. Neuro Oncol. 20 (Suppl 4):iv1–iv86. 2018. View Article : Google Scholar : PubMed/NCBI
3	Patel NP, Lyon KA and Huang JH: The effect of race on the prognosis of the glioblastoma patient: A brief review. Neurol Res. 41:967–971. 2019. View Article : Google Scholar : PubMed/NCBI
4	Taphoorn MJ, Sizoo EM and Bottomley A: Review on quality of life issues in patients with primary brain tumors. Oncologist. 15:618–626. 2010. View Article : Google Scholar : PubMed/NCBI
5	Bartel DP: MicroRNAs: Target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI
6	Pasquinelli AE: MicroRNAs and their targets: Recognition, regulation and an emerging reciprocal relationship. Nat Rev Genet. 13:271–282. 2012. View Article : Google Scholar : PubMed/NCBI
7	Li XT, Wang HZ, Wu ZW, Yang TQ, Zhao ZH, Chen GL, Xie XS, Li B, Wei YX, Huang YL, et al: miR-494-3p regulates cellular proliferation, invasion, migration, and apoptosis by PTEN/AKT signaling in human glioblastoma cells. Cell Mol Neurobiol. 35:679–687. 2015. View Article : Google Scholar : PubMed/NCBI
8	Chae DK, Park J, Cho M, Ban E, Jang M, Yoo YS, Kim EE, Baik JH and Song EJ: MiR-195 and miR-497 suppress tumorigenesis in lung cancer by inhibiting SMURF2-induced TGF-β receptor I ubiquitination. Mol Oncol. 13:2663–2678. 2019. View Article : Google Scholar : PubMed/NCBI
9	Shen J and Li M: MicroRNA-744 inhibits cellular proliferation and invasion of colorectal cancer by directly targeting oncogene notch1. Oncol Res. 26:1401–1409. 2018. View Article : Google Scholar : PubMed/NCBI
10	Chen XF and Liu Y: MicroRNA-744 inhibited cervical cancer growth and progression through apoptosis induction by regulating Bcl-2. Biomed Pharmacother. 81:379–387. 2016. View Article : Google Scholar : PubMed/NCBI
11	Wang J, Cai H, Dai Z and Wang G: Down-regulation of lncRNA XIST inhibits cell proliferation via regulating miR-744/RING1 axis in non-small cell lung cancer. Clin Sci (Lond). 133:1567–1579. 2019. View Article : Google Scholar : PubMed/NCBI
12	Guo Q, Zhang Q, Lu L and Xu Y: Long noncoding RNA RUSC1-AS1 promotes tumorigenesis in cervical cancer by acting as a competing endogenous RNA of microRNA-744 and consequently increasing Bcl-2 expression. Cell Cycle. 19:1222–1235. 2020. View Article : Google Scholar : PubMed/NCBI
13	Miyamae M, Komatsu S, Ichikawa D, Kawaguchi T, Hirajima S, Okajima W, Ohashi T, Imamura T, Konishi H and Shiozaki A: Plasma microRNA profiles: Identification of miR-744 as a novel diagnostic and prognostic biomarker in pancreatic cancer. Br J Cancer. 113:1467–1476. 2015. View Article : Google Scholar : PubMed/NCBI
14	Kleemann M, Schneider H, Unger K, Sander P, Schneider EM, Fischer-Posovszky P, Handrick R and Otte K: MiR-744-5p inducing cell death by directly targeting HNRNPC and NFIX in ovarian cancer cells. Sci Rep. 8:90202018. View Article : Google Scholar : PubMed/NCBI
15	Chen S, Shi F, Zhang W, Zhou Y and Huang J: miR-744-5p inhibits non-small cell lung cancer proliferation and invasion by directly targeting PAX2. Technol Cancer Res Treat. 18:15330338198769132019. View Article : Google Scholar : PubMed/NCBI
16	Hübner M, Hinske CL, Effinger D, Wu T, Thon N, Kreth FW and Kreth S: Intronic miR-744 inhibits glioblastoma migration by functionally antagonizing its host gene MAP2K4. Cancers (Basel). 25:4002018. View Article : Google Scholar
17	Deng Y, Li Y, Fang Q, Luo H and Zhu G: microRNA-744 is downregulated in glioblastoma and inhibits the aggressive behaviors by directly targeting NOB1. Am J Cancer Res. 8:2238–2253. 2018.PubMed/NCBI
18	Li Y, Gan S, Ren L, Yuan L, Liu J, Wang W, Wang X, Zhang Y, Jiang J, Zhang F and Qi X: Multifaceted regulation and functions of replication factor C family in human cancers. Am J Cancer Res. 8:1343–1355. 2018.PubMed/NCBI
19	Xiong S, Wang Q, Zheng L, Gao F and Li J: Identification of candidate molecular markers of nasopharyngeal carcinoma by tissue microarray and in situ hybridization. Med Oncol. 28 (Suppl 1):S341–S348. 2011. View Article : Google Scholar : PubMed/NCBI
20	Xu J, Wang G, Gong W, Guo S, Li D and Zhan Q: The noncoding function of NELFA mRNA promotes the development of oesophageal squamous cell carcinoma by regulating the Rad17-RFC2-5 complex. Mol Oncol. 14:611–624. 2020. View Article : Google Scholar : PubMed/NCBI
21	Ho KH, Kuo TC, Lee YT, Chen PH, Shih CM, Cheng CH, Liu AJ, Lee CC and Chen KC: Xanthohumol regulates miR-4749-5p-inhibited RFC2 signaling in enhancing temozolomide cytotoxicity to glioblastoma. Life Sci. 254:1178072020. View Article : Google Scholar : PubMed/NCBI
22	Hu T, Shen H, Li J, Yang P, Gu Q and Fu Z: RFC2, a direct target of miR-744, modulates the cell cycle and promotes the proliferation of CRC cells. J Cell Physiol. 235:8319–8333. 2020. View Article : Google Scholar : PubMed/NCBI
23	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. View Article : Google Scholar : PubMed/NCBI
24	Kato T, Natsume A, Toda H, Iwamizu H, Sugita T, Hachisu R, Watanabe R, Yuki K, Motomura K, Bankiewicz K and Wakabayashi T: Efficient delivery of liposome-mediated MGMT-siRNA reinforces the cytotoxity of temozolomide in GBM-initiating cells. Gene Ther. 17:1363–1371. 2010. View Article : Google Scholar : PubMed/NCBI
25	Omuro A and DeAngelis LM: Glioblastoma and other malignant gliomas: A clinical review. Jama. 310:1842–1850. 2013. View Article : Google Scholar : PubMed/NCBI
26	Gilbert MR, Wang M, Aldape KD, Stupp R, Hegi ME, Jaeckle KA, Armstrong TS, Wefel JS, Won M, Blumenthal DT, et al: Dose-dense temozolomide for newly diagnosed glioblastoma: A randomized phase III clinical trial. J Clin Oncol. 31:4085–4091. 2013. View Article : Google Scholar : PubMed/NCBI
27	Cloughesy TF, Cavenee WK and Mischel PS: Glioblastoma: From molecular pathology to targeted treatment. Ann Rev Pathol. 9:1–25. 2014. View Article : Google Scholar : PubMed/NCBI
28	Johnson DR and O'Neill BP: Glioblastoma survival in the United States before and during the temozolomide era. J Neurooncol. 107:359–364. 2012. View Article : Google Scholar : PubMed/NCBI
29	Luo JW, Wang X, Yang Y and Mao Q: Role of micro-RNA (miRNA) in pathogenesis of glioblastoma. Eur Rev Med Pharmacol Sci. 19:1630–1639. 2015.PubMed/NCBI
30	Saadatpour L, Fadaee E, Fadaei S, Mansour RN, Mohammadi M, Mousavi SM, Goodarzi M, Verdi J and Mirzaei H: Glioblastoma: Exosome and microRNA as novel diagnosis biomarkers. Cancer Gene Ther. 23:415–418. 2016. View Article : Google Scholar : PubMed/NCBI
31	Cui JQ, Shi YF and Zhou HJ: Expression of RFC2 and PCNA in different gestational trophoblastic diseases. Ai Zheng. 23:196–200. 2004.(In Chinese). PubMed/NCBI
32	Olbrich K, Costard L, Möser CV, Syhr KMJ, King-Himmelreich TS, Wolters MC, Schmidtko A, Geisslinger G and Niederberger E: Cleavage of SNAP-25 ameliorates cancer pain in a mouse model of melanoma. Eur J Pain. 21:101–111. 2017. View Article : Google Scholar : PubMed/NCBI
33	Mu Y, Yan X, Li D, Zhao D, Wang L, Wang X, Gao D, Yang J, Zhang H, Li Y, et al: NUPR1 maintains autolysosomal efflux by activating SNAP25 transcription in cancer cells. Autophagy. 14:654–670. 2018. View Article : Google Scholar : PubMed/NCBI
34	Antonucci F, Corradini I, Morini R, Fossati G, Menna E, Pozzi D, Pacioni S, Verderio C, Bacci A and Matteoli M: Reduced SNAP-25 alters short-term plasticity at developing glutamatergic synapses. EMBO Rep. 14:645–651. 2013. View Article : Google Scholar : PubMed/NCBI
35	Fulda S, Wick W, Weller M and Debatin KM: Smac agonists sensitize for Apo2L/TRAIL- or anticancer drug-induced apoptosis and induce regression of malignant glioma in vivo. Nat Med. 8:808–815. 2002. View Article : Google Scholar : PubMed/NCBI
36	Shiraishi S, Tada K, Nakamura H, Makino K, Kochi M, Saya H, Kuratsu Ji and Ushio Y: Influence of p53 mutations on prognosis of patients with glioblastoma. Cancer. 95:249–257. 2002. View Article : Google Scholar : PubMed/NCBI
37	Wu T and Dai Y: Tumor microenvironment and therapeutic response. Cancer Lett. 387:61–68. 2017. View Article : Google Scholar : PubMed/NCBI
38	She X, Yu Z, Cui Y, Lei Q, Wang Z, Xu G, Xiang J, Wu M and Li G: miR-128 and miR-149 enhance the chemosensitivity of temozolomide by Rap1B-mediated cytoskeletal remodeling in glioblastoma. Oncol Rep. 32:957–964. 2014. View Article : Google Scholar : PubMed/NCBI
39	Xu P, Yang J, Liu J, Yang X, Liao J, Yuan F, Xu Y, Liu B and Chen Q: Identification of glioblastoma gene prognosis modules based on weighted gene co-expression network analysis. BMC Med Genomics. 11:962018. View Article : Google Scholar : PubMed/NCBI
40	Xiang Y, Zhang CQ and Huang K: Predicting glioblastoma prognosis networks using weighted gene co-expression network analysis on TCGA data. BMC Bioinformatics. 13 (Suppl 2):S122012. View Article : Google Scholar : PubMed/NCBI
41	Kong Y, Feng ZC, Zhang YL, Liu XF, Ma Y, Zhao ZM, Huang B, Chen AJ, Zhang D, Thorsen F, et al: Identification of immune-related genes contributing to the development of glioblastoma using weighted gene co-expression network analysis. Front Immunol. 11:12812020. View Article : Google Scholar : PubMed/NCBI