lncRNA NEAT1 regulates the proliferation and migration of hepatocellular carcinoma cells by acting as a miR‑320a molecular sponge and targeting L antigen family member 3

Zhang,Yixi; Huang,Changjun; Zhu,Zebin; Hou,Yufei; Huang,Shanzhou; Sun,Chengjun; Tang,Yunhua; Zhang,Zhiheng; Wang,Linhe; Chen,Huadi; Ju,Weiqiang; Qiao,Xin; Chen,Maogen

doi:10.3892/ijo.2020.5108

lncRNA NEAT1 regulates the proliferation and migration of hepatocellular carcinoma cells by acting as a miR‑320a molecular sponge and targeting L antigen family member 3

Authors:
- Yixi Zhang
- Changjun Huang
- Zebin Zhu
- Yufei Hou
- Shanzhou Huang
- Chengjun Sun
- Yunhua Tang
- Zhiheng Zhang
- Linhe Wang
- Huadi Chen
- Weiqiang Ju
- Xin Qiao
- Maogen Chen
View Affiliations

Affiliations: Organ Transplant Center, The First Affiliated Hospital, Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China, Organ Transplant Center, The First Affiliated Hospital, Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China, Department of Ultrasound Medicine, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, Guangdong 510600, P.R. China, Department of Anesthesiology, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, P.R. China
Published online on: August 10, 2020 https://doi.org/10.3892/ijo.2020.5108
Pages: 1001-1012

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

Long non‑coding RNAs (lncRNAs) serve a pivotal role in hepatocellular carcinoma (HCC) progression and have been confirmed to participate in the carcinogenesis and development of HCC. Certain studies have focused on lncRNA nuclear enriched abundant transcript 1 (NEAT1) in HCC. However, the relationship between lncRNA NEAT1 and HCC remains unclear. The present study found that NEAT1 was significantly overexpressed in HCC cell lines compared with LX‑2 hepatic stellate cells. NEAT1 expression in Huh7 and MHCC‑97H cells was increased following transfection with lentivirus (LV)‑NEAT1 but inhibited by LV‑short hairpin NEAT1. Knockdown of NEAT1 significantly repressed HCC cell viability, increased cell apoptosis, and inhibited cell migration and invasion capacity. By contrast, upregulation of NEAT1 demonstrated the reverse effects. Furthermore, microRNA‑320a (miR‑230a) was predicted to be a direct target of NEAT1 and was significantly reduced in HCC cells. Additionally, a luciferase activity reporter assay and RNA immunoprecipitation assay were performed to confirm the interaction between miR‑320a and NEAT1. Using a dual‑luciferase activity assay, L antigen family member 3 (LAGE3) was found to be a target of miR‑320a. Finally, in vivo nude mouse models were established, and the results indicated that NEAT1 suppressed HCC progression by targeting miR‑320a. In conclusion, the present findings revealed that the NEAT1/miR‑320a/LAGE3 axis participates in HCC development and that NEAT1 could be used as a biomarker for HCC.

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 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Sia D, Villanueva A, Friedman SL and Llovet JM: Liver cancer cell of origin, molecular class, and effects on patient prognosis. Gastroenterology. 152:745–761. 2017. View Article : Google Scholar : PubMed/NCBI
3	Villanueva A: Hepatocellular carcinoma. N Engl J Med. 380:1450–1462. 2019. View Article : Google Scholar : PubMed/NCBI
4	Xu J: Trends in liver cancer mortality among adults aged 25 and over in the United States, 2000-2016. NCHS Data Brief. 1–8. 2018.
5	Necsulea A, Soumillon M, Warnefors M, Liechti A, Daish T, Zeller U, Baker JC, Grützner F and Kaessmann H: The evolution of lncRNA repertoires and expression patterns in tetrapods. Nature. 505:635–640. 2014. View Article : Google Scholar : PubMed/NCBI
6	Fernandes JCR, Acuña SM, Aoki JI, Floeter-Winter LM and Muxel SM: Long non-coding RNAs in the regulation of gene expression: Physiology and disease. Noncoding RNA. 5:172019.
7	Bartel DP: Metazoan MicroRNAs. Cell. 173:20–51. 2018. View Article : Google Scholar : PubMed/NCBI
8	Friedman RC, Farh KKH, Burge CB and Bartel DP: Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19:92–105. 2009. View Article : Google Scholar
9	Kabekkodu SP, Shukla V, Varghese VK, D' Souza J, Chakrabarty S and Satyamoorthy K: Clustered miRNAs and their role in biological functions and diseases. Biol Rev. 93:1955–1986. 2018. View Article : Google Scholar : PubMed/NCBI
10	Marchese FP, Raimondi I and Huarte M: The multidimensional mechanisms of long noncoding RNA function. Genome Biol. 18:2062017. View Article : Google Scholar : PubMed/NCBI
11	Ding L, Ren J, Zhang D, Li Y, Huang X, Hu Q, Wang H, Song Y, Ni Y and Hou Y: A novel stromal lncRNA signature reprograms fibroblasts to promote the growth of oral squamous cell carcinoma via LncRNA-CAF/interleukin-33. Carcinogenesis. 39:397–406. 2018. View Article : Google Scholar : PubMed/NCBI
12	Xing F, Liu Y, Wu SY, Wu K, Sharma S, Mo YY, Feng J, Sanders S, Jin G, Singh R, et al: Loss of XIST in breast cancer activates MSN-c-Met and reprograms microglia via exosomal miRNA to promote brain metastasis. Cancer Res. 78:4316–4330. 2018. View Article : Google Scholar : PubMed/NCBI
13	Yang Y, Zhang JP, Chen X, Xu X, Cao G, Li H and Wu T: LncRNA FTX sponges miR-215 and inhibits phosphorylation of vimentin for promoting colorectal cancer progression. Gene Ther. 25:321–330. 2018. View Article : Google Scholar : PubMed/NCBI
14	Chen X, Chen Z, Yu S, Nie F, Yan S, Ma P, Chen Q, Wei C, Fu H, Xu T, et al: Long noncoding RNA LINC01234 functions as a competing endogenous RNA to regulate CBFB expression by sponging miR-204-5p in gastric cancer. Clin Cancer Res. 24:2002–2014. 2018. View Article : Google Scholar : PubMed/NCBI
15	Zhao L, Zhang YJ and Zhang YB: Long noncoding RNA CASC2 regulates hepatocellular carcinoma cell oncogenesis through miR-362-5p/Nf-B axis. J Cell Physiol. 233:6661–6670. 2018. View Article : Google Scholar : PubMed/NCBI
16	Yang X, Qu S, Wang L, Zhang H, Yang Z, Wang J, Dai B, Tao K, Shang R, Liu Z, et al: PTBP3 splicing factor promotes hepatocellular carcinoma by destroying the splicing balance of NEAT1 and pre-miR-612. Oncogene. 37:6399–6413. 2018. View Article : Google Scholar : PubMed/NCBI
17	Liu Z, Chang Q, Yang F, Liu B, Yao HW, Bai ZG, Pu CS, Ma XM, Yang Y, Wang TT, et al: Long non-coding RNA NEAT1 overexpression is associated with unfavorable prognosis in patients with hepatocellular carcinoma after hepatectomy: A Chinese population-based study. Eur J Surg Onc. 43:1697–1703. 2017. View Article : Google Scholar
18	Chen S and Xia X: Long noncoding RNA NEAT1 suppresses sorafenib sensitivity of hepatocellular carcinoma cells via regu-lating miR-335-c-Met. J Cell Physiol. Apr 1–2019.Epub ahead of print.
19	Müller V, Oliveira-Ferrer L, Steinbach B, Pantel K and Schwarzenbach H: Interplay of lncRNA H19/miR-675 and lncRNA NEAT1/miR-204 in breast cancer. Mol Oncol. 13:1137–1149. 2019. View Article : Google Scholar : PubMed/NCBI
20	Kong X, Zhao Y, Li X, Tao Z, Hou M and Ma H: Overexpression of HIF-2α-dependent NEAT1 promotes the progression of non-small cell lung cancer through miR-101-3p/SOX9/Wnt/β-catenin signal pathway. Cell Physiol Biochem. 52:368–381. 2019. View Article : Google Scholar
21	Zhang M, Weng WW, Zhang QY, Wu Y, Ni S, Tan C, Xu M, Sun H, Liu C, Wei P and Du X: The lncRNA NEAT1 activates Wnt/β-catenin signaling and promotes colorectal cancer progression via interacting with DDX5. J Hematol Oncol. 11:1132018. View Article : Google Scholar
22	Yan P, Su Z, Zhang Z and Gao T: LncRNA NEAT1 enhances the resistance of anaplastic thyroid carcinoma cells to cisplatin by sponging miR-9-5p and regulating SPAG9 expression. Int J Oncol. 55:988–1002. 2019.PubMed/NCBI
23	Xia TF, Chen J, Wu K, Zhang J and Yan Q: Long noncoding RNA NEAT1 promotes the growth of gastric cancer cells by regulating miR-497-5p/PIK3R1 axis. Eur Rev Med Pharmacol Sci. 23:6914–6926. 2019.PubMed/NCBI
24	He J, Xu F, Man X, Zhang Y and Li H: Long non-coding RNA NEAT1 promotes tumor development and metastasis through targeting RAB9A in malignant melanoma. Minerva Med. Jul 17–2019. View Article : Google Scholar : Online ahead of print. PubMed/NCBI
25	Livak KJ and Schmittgen TD: Analysis of relative gene expres-sion data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
26	Sun JY, Zhao ZW, Li WM, Yang G, Jing PY, Li P, Dang HZ, Chen Z, Zhou YA and Li XF: Knockdown of MALAT1 expression inhibits HUVEC proliferation by upregulation of miR-320a and downregulation of FOXM1 expression. Oncotarget. 8:61499–61509. 2017. View Article : Google Scholar : PubMed/NCBI
27	Scholzen T and Gerdes J: The Ki-67 protein: From the known and the unknown. J Cell Physiol. 182:311–322. 2000. View Article : Google Scholar : PubMed/NCBI
28	Brown DC and Gatter KC: Ki67 protein: The immaculate deception? Histopathology. 40:2–11. 2002. View Article : Google Scholar : PubMed/NCBI
29	Ponting CP, Oliver PL and Reik W: Evolution and functions of long noncoding RNAs. Cell. 136:629–641. 2009. View Article : Google Scholar : PubMed/NCBI
30	Clemson CM, Hutchinson JN, Sara SA, Ensminger AW, Fox AH, Chess A and Lawrence JB: An architectural role for a nuclear noncoding RNA: NEAT1 RNA is essential for the structure of paraspeckles. Mol Cell. 33:717–726. 2009. View Article : Google Scholar : PubMed/NCBI
31	Souquere S, Beauclair G, Harper F, Fox A and Pierron G: Highly ordered spatial organization of the structural long noncoding NEAT1 RNAs within paraspeckle nuclear bodies. Mol Biol Cell. 21:4020–4027. 2010. View Article : Google Scholar : PubMed/NCBI
32	Fortunato O, Borzi C, Milione M, Centonze G, Conte D, Boeri M, Verri C, Moro M, Facchinetti F, Andriani F, et al: Circulating mir-320a promotes immunosuppressive macrophages M2 phenotype associated with lung cancer risk. Int J Cancer. 144:2746–2761. 2019. View Article : Google Scholar
33	Sun L, Liu B, Lin Z, Yao Y, Chen Y, Li Y, Chen J, Yu D, Tang Z, Wang B, et al: miR-320a acts as a prognostic factor and Inhibits metastasis of salivary adenoid cystic carcinoma by targeting ITGB3. Mol Cancer. 14:962015. View Article : Google Scholar : PubMed/NCBI
34	Yu J, Wang JG, Zhang L, Yang HP, Wang L, Ding D, Chen Q, Yang WL, Ren KH, Zhou DM, et al: MicroRNA-320a inhibits breast cancer metastasis by targeting metadherin. Oncotarget. 7:38612–38625. 2016. View Article : Google Scholar : PubMed/NCBI
35	Shu S, Liu X, Xu M, Gao X, Chen S, Zhang L and Li R: MicroRNA-320a acts as a tumor suppressor in endometrial carcinoma by targeting IGF-1R. Int J Mol Med. 43:1505–1512. 2019.PubMed/NCBI
36	Zhang Z, Li X, Sun W, Yue S, Yang J, Li J, Ma B, Wang J, Yang X, Pu M, et al: Loss of exosomal miR-320a from cancer-associated fibroblasts contributes to HCC proliferation and metastasis. Cancer Lett. 397:33–42. 2017. View Article : Google Scholar : PubMed/NCBI
37	Alpen B, Güre AO, Scanlan MJ, Old LJ and Chen YT: A new member of the NY-ESO-1 gene family is ubiquitously expressed in somatic tissues and evolutionarily conserved. Gene. 297:141–149. 2002. View Article : Google Scholar : PubMed/NCBI
38	Wan LC, Maisonneuve P, Szilard RK, Lambert JP, Ng TF, Manczyk N, Huang H, Laister R, Caudy AA, Gingras AC, et al: Proteomic analysis of the human KEOPS complex identifies C14ORF142 as a core subunit homologous to yeast Gon7. Nucleic Acids Res. 45:805–817. 2017. View Article : Google Scholar
39	Daugeron MC, Lenstra TL, Frizzarin M, El Yacoubi B, Liu X, Baudin-Baillieu A, Lijnzaad P, Decourty L, Saveanu C, Jacquier A, et al: Gcn4 misregulation reveals a direct role for the evolutionary conserved EKC/KEOPS in the t6A modification of tRNAs. Nucleic Acids Res. 39:6148–6160. 2011. View Article : Google Scholar : PubMed/NCBI
40	Rojas-Benitez D, Ibar C and Glavic A: The Drosophila EKC/KEOPS complex: Roles in protein synthesis homeostasis and animal growth. Fly (Austin). 7:168–172. 2013. View Article : Google Scholar
41	Costessi A, Mahrour N, Sharma V, Stunnenberg R, Stoel MA, Tijchon E, Conaway JW, Conaway RC and Stunnenberg HG: The human EKC/KEOPS complex is recruited to Cullin2 ubiquitin ligases by the human tumour antigen PRAME. PLoS One. 7:e428222012. View Article : Google Scholar : PubMed/NCBI
42	Liu S, Liu LH, Hu WW and Wang M: Long noncoding RNA TUG1 regulates the development of oral squamous cell carcinoma through sponging miR-524-5p to mediate DLX1 expression as a competitive endogenous RNA. J Cell Physiol. 234:20206–20216. 2019. View Article : Google Scholar : PubMed/NCBI