Long non‑coding RNA maternally expressed gene 3 affects cell proliferation, apoptosis and migration by targeting the microRNA‑9‑5p/midkine axis and activating the phosphoinositide‑dependent kinase/AKT pathway in hepatocellular carcinoma

Wu,Dezhi; Ma,Zheng; Ma,Deyu; Li,Qiquan

doi:10.3892/ol.2021.12606

Long non‑coding RNA maternally expressed gene 3 affects cell proliferation, apoptosis and migration by targeting the microRNA‑9‑5p/midkine axis and activating the phosphoinositide‑dependent kinase/AKT pathway in hepatocellular carcinoma

Authors:
- Dezhi Wu
- Zheng Ma
- Deyu Ma
- Qiquan Li
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Affiliations: School of Pharmaceutical Sciences, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou 550002, P.R. China, Faculty of Pharmaceutical Sciences, Cangzhou Central Hospital, Cangzhou, Hebei 061001, P.R. China, Department of Pharmacy, Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou, Sichuan 646000, P.R. China, GCP Center, Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou, Sichuan 646000, P.R. China
Published online on: March 3, 2021 https://doi.org/10.3892/ol.2021.12606
Article Number: 345
Copyright: © Wu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Long non‑coding RNA (lncRNA) maternally expressed gene 3 (MEG3) is a tumor suppressor in several cancers, such as glioma, prostate cancer and esophageal cancer. However, the role of MEG3 in hepatocellular carcinoma (HCC) and the related molecular mechanisms are not well understood. The present study aimed to determine the biological function of MEG3 in regulating HCC cell viability, apoptosis and migration. In addition, the interaction between MEG3, microRNA (miR)‑9‑5p and Midkine (MDK), and the activation of the phosphoinositide‑dependent kinase (PDK)/AKT pathway in HCC cell line MHCC‑97L were examined. Luciferase reporter assays, reverse transcription‑quantitative PCR and western blotting were used to determine the interaction between MEG3, miR‑9‑5p and MDK and the activation of the PDK/AKT pathway. Cell viability was determined by the CCK8 assay and the cell cycle analysis using flow cytometry analysis. Cell apoptosis was examined by flow cytometry analysis and caspase 3/9 activity. Wound healing assays and western blotting were used to investigate cell migration. The present study demonstrated that MEG3 suppressed HCC cell viability and migration, and induced cell apoptosis. In addition, it was also found that MEG3 targets the miR‑9‑5p/MDK axis and modulates the PDK/AKT pathway in HCC. In conclusion, the findings of the present study demonstrated that lncRNA MEG3 affects HCC cell viability, apoptosis and migration through its targeting of miR‑9‑5p/MDK and regulation of the PDK/AKT pathway. The MEG3/miR‑9‑5p/MDK axis may be a potential therapeutic target in HCC.

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1	McGlynn KA, Petrick JL and London WT: Global epidemiology of hepatocellular carcinoma: An emphasis on demographic and regional variability. Clin Liver Dis. 19:223–238. 2015. View Article : Google Scholar : PubMed/NCBI
2	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2018. CA Cancer J Clin. 68:7–30. 2018. View Article : Google Scholar : PubMed/NCBI
3	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
4	Korean Liver Cancer Association, National Cancer Center, . 2018 Korean Liver Cancer Association-National Cancer Center Korea Practice Guidelines for the Management of Hepatocellular Carcinoma. Gut Liver. 13:227–299. 2019. View Article : Google Scholar : PubMed/NCBI
5	Bhan A, Soleimani M and Mandal SS: Long noncoding RNA and cancer: A new paradigm. Cancer Res. 77:3965–3981. 2017. View Article : Google Scholar : PubMed/NCBI
6	Zhang Y, Liu J, Lv Y, Zhang C and Guo S: lncRNA meg3 suppresses hepatocellular carcinoma in vitro and vivo studies. Am J Transl Res. 11:4089–4099. 2019.PubMed/NCBI
7	Zhuo H, Tang J, Lin Z, Jiang R, Zhang X, Ji J, Wang P and Sun B: The aberrant expression of MEG3 regulated by UHRF1 predicts the prognosis of hepatocellular carcinoma. Mol Carcinog. 55:209–219. 2016. View Article : Google Scholar : PubMed/NCBI
8	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
9	Wu F, Yang Z and Li G: Role of specific microRNAs for endothelial function and angiogenesis. Biochem Biophys Res Commun. 386:549–553. 2009. View Article : Google Scholar : PubMed/NCBI
10	Lopez-Urrutia E, Bustamante Montes LP, Ladron de Guevara Cervantes D, Perez-Plasencia C and Campos-Parra AD: Crosstalk between long non-coding RNAs, Micro-RNAs and mRNAs: Deciphering molecular mechanisms of master regulators in cancer. Front Oncol. 9:6692019. View Article : Google Scholar : PubMed/NCBI
11	Tam C, Wong JH, Tsui SKW, Zuo T, Chan TF and Ng TB: lncRNAs with miRNAs in regulation of gastric, liver, and colorectal cancers: Updates in recent years. Appl Microbiol Biotechnol. 103:4649–4677. 2019. View Article : Google Scholar : PubMed/NCBI
12	Snezhkina AV, Krasnov GS, Zhikrivetskaya SO, Karpova IY, Fedorova MS, Nyushko KM, Belyakov MM, Gnuchev NV, Sidorov DV, Alekseev BY, et al: Overexpression of microRNAs miR-9, −98, and −199 correlates with the downregulation of HK2 expression in colorectal cancer. Mol Biol (Mosk). 52:220–230. 2018.(In Russian). View Article : Google Scholar : PubMed/NCBI
13	Zhou L, Fu L, Lv N, Chen XS, Liu J, Li Y, Xu QY, Huang S, Zhang XD, Dou LP, et al: A minicircuitry comprised of microRNA-9 and SIRT1 contributes to leukemogenesis in t(8;21) acute myeloid leukemia. Eur Rev Med Pharmacol Sci. 21:786–794. 2017.PubMed/NCBI
14	Tsai KW, Liao YL, Wu CW, Hu LY, Li SC, Chan WC, Ho MR, Lai CH, Kao HW, Fang WL, et al: Aberrant hypermethylation of miR-9 genes in gastric cancer. Epigenetics. 6:1189–1197. 2011. View Article : Google Scholar : PubMed/NCBI
15	Wu M, Huang Y, Chen T, Wang W, Yang S, Ye Z and Xi X: lncRNA MEG3 inhibits the progression of prostate cancer by modulating miR-9-5p/QKI-5 axis. J Cell Mol Med. 23:29–38. 2019. View Article : Google Scholar : PubMed/NCBI
16	Dong Z, Zhang A, Liu S, Lu F, Guo Y, Zhang G, Xu F, Shi Y, Shen S, Liang J and Guo W: Aberrant methylation-mediated silencing of lncRNA MEG3 functions as a ceRNA in esophageal cancer. Mol Cancer Res. 15:800–810. 2017. View Article : Google Scholar : PubMed/NCBI
17	Kretschmer PJ, Fairhurst JL, Decker MM, Chan CP, Gluzman Y, Böhlen P and Kovesdi I: Cloning, characterization and developmental regulation of two members of a novel human gene family of neurite outgrowth-promoting proteins. Growth Factors. 5:99–114. 1991. View Article : Google Scholar : PubMed/NCBI
18	Filippou PS, Karagiannis GS and Constantinidou A: Midkine (MDK) growth factor: A key player in cancer progression and a promising therapeutic target. Oncogene. 39:2040–2054. 2020. View Article : Google Scholar : PubMed/NCBI
19	Wang F, Shan S, Huo Y, Xie Z, Fang Y, Qi Z, Chen F, Li Y and Sun B: miR-155-5p inhibits PDK1 and promotes autophagy via the mTOR pathway in cervical cancer. Int J Biochem Cell Biol. 99:91–99. 2018. View Article : Google Scholar : PubMed/NCBI
20	Meng F, Zhang Y, Li X, Wang J and Wang Z: Clinical significance of miR-138 in patients with malignant melanoma through targeting of PDK1 in the PI3K/AKT autophagy signaling pathway. Oncol Rep. 38:1655–1662. 2017. View Article : Google Scholar : PubMed/NCBI
21	Chuang CK, Lin HCA, Tasi HY, Lee KH, Kao Y, Chuang FL, Chang YH, Lin PH, Liu CY and Pang ST: Clinical presentations and molecular studies of invasive renal epithelioid angiomyolipoma. Int Urol Nephrol. 49:1527–1536. 2017. View Article : Google Scholar : PubMed/NCBI
22	Lu J, Liu QH, Wang F, Tan JJ, Deng YQ, Peng XH, Liu X, Zhang B, Xu X and Li XP: Exosomal miR-9 inhibits angiogenesis by targeting MDK and regulating PDK/AKT pathway in nasopharyngeal carcinoma. J Exp Clin Cancer Res. 37:1472018. 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	Abbastabar M, Sarfi M, Golestani A and Khalili E: lncRNA involvement in hepatocellular carcinoma metastasis and prognosis. EXCLI J. 17:900–913. 2018.PubMed/NCBI
25	Huang Z, Zhou JK, Peng Y, He W and Huang C: The role of long noncoding RNAs in hepatocellular carcinoma. Mol Cancer. 19:772020. View Article : Google Scholar : PubMed/NCBI
26	Wei GH and Wang X: lncRNA MEG3 inhibit proliferation and metastasis of gastric cancer via p53 signaling pathway. Eur Rev Med Pharmacol Sci. 21:3850–3856. 2017.PubMed/NCBI
27	Zhang CY, Yu MS, Li X, Zhang Z, Han CR and Yan B: Overexpression of long non-coding RNA MEG3 suppresses breast cancer cell proliferation, invasion, and angiogenesis through AKT pathway. Tumour Biol. 39:10104283177013112017.PubMed/NCBI
28	Zhou Y, Zhang X and Klibanski A: MEG3 noncoding RNA: A tumor suppressor. J Mol Endocrinol. 48:R45–R53. 2012. View Article : Google Scholar : PubMed/NCBI
29	He JH, Han ZP, Liu JM, Zhou JB, Zou MX, Lv YB, Li YG and Cao MR: Overexpression of long non-coding RNA meg3 inhibits proliferation of hepatocellular carcinoma Huh7 Cells via negative modulation of miRNA-664. J Cell Biochem. 118:3713–3721. 2017. View Article : Google Scholar : PubMed/NCBI
30	Liu H, Zhou Y and Tang L: Caffeine induces sustained apoptosis of human gastric cancer cells by activating the caspase-9/caspase-3 signalling pathway. Mol Med Rep. 16:2445–2454. 2017. View Article : Google Scholar : PubMed/NCBI
31	Altadill A, Rodríguez M, González LO, Junquera S, Corte MD, González-Dieguez ML, Linares A, Barbón E, Fresno-Forcelledo M, Rodrigo L and Vizoso FJ: Liver expression of matrix metalloproteases and their inhibitors in hepatocellular carcinoma. Dig Liver Dis. 41:740–748. 2009. View Article : Google Scholar : PubMed/NCBI
32	Veneziano D, Marceca GP, Di Bella S, Nigita G, Distefano R and Croce CM: Investigating miRNA-lncRNA interactions: Computational tools and resources. Methods Mol Biol. 1970:251–277. 2019. View Article : Google Scholar : PubMed/NCBI
33	Wang X, Su R, Guo Q, Liu J, Ruan B and Wang G: Competing endogenous RNA (ceRNA) hypothetic model based on comprehensive analysis of long non-coding RNA expression in lung adenocarcinoma. PeerJ. 7:e80242019. View Article : Google Scholar : PubMed/NCBI
34	Huang Y: The novel regulatory role of lncRNA-miRNA-mRNA axis in cardiovascular diseases. J Cell Mol Med. 22:5768–5775. 2018. View Article : Google Scholar : PubMed/NCBI
35	Chen K, Ma Y, Wu S, Zhuang Y, Liu X, Lv L and Zhang G: Construction and analysis of a lncRNA-miRNA-mRNA network based on competitive endogenous RNA reveals functional lncRNAs in diabetic cardiomyopathy. Mol Med Rep. 20:1393–1403. 2019.PubMed/NCBI
36	Zhu D, Xiao Z, Wang Z, Hu B, Duan C, Zhu Z, Gao N, Zhu Y and Wang H: MEG3/MIR-376B-3P/HMGA2 axis is involved in pituitary tumor invasiveness. J Neurosurg. 2020.(Ahead of print).
37	Lu WX: Long non-coding RNA MEG3 represses cholangiocarcinoma by regulating miR-361-5p/TRAF3 axis. Eur Rev Med Pharmacol Sci. 23:7356–7368. 2019.PubMed/NCBI
38	Zhang S and Guo W: Long non-coding RNA MEG3 suppresses the growth of glioma cells by regulating the miR-96-5p/MTSS1 signaling pathway. Mol Med Rep. 20:4215–4225. 2019.PubMed/NCBI
39	Shen X, Bai H, Zhu H, Yan Q, Yang Y, Yu W, Shi Q, Wang J, Li J and Chen L: Long non-coding RNA MEG3 functions as a competing endogenous RNA to regulate HOXA11 expression by sponging miR-181a in multiple myeloma. Cell Physiol Biochem. 49:87–100. 2018. View Article : Google Scholar : PubMed/NCBI
40	Liu Z, Chen JY, Zhong Y, Xie L and Li JS: lncRNA MEG3 inhibits the growth of hepatocellular carcinoma cells by sponging miR-9-5p to upregulate SOX11. Braz J Med Biol Res. 52:e86312019. View Article : Google Scholar : PubMed/NCBI
41	Li XY, Wen JY, Jia CC, Wang TT, Li X, Dong M, Lin QU, Chen ZH, Ma XK, Wei LI, et al: MicroRNA-34a-5p enhances sensitivity to chemotherapy by targeting AXL in hepatocellular carcinoma MHCC-97L cells. Oncol Lett. 10:2691–2698. 2015. View Article : Google Scholar : PubMed/NCBI
42	Kai AK, Chan LK, Lo RC, Lee JM, Wong CC, Wong JC and Ng IO: Down-regulation of TIMP2 by HIF-1α/miR-210/HIF-3α regulatory feedback circuit enhances cancer metastasis in hepatocellular carcinoma. Hepatology. 64:473–487. 2016. View Article : Google Scholar : PubMed/NCBI