microRNA-29a functions as a tumor suppressor in nasopharyngeal carcinoma 5-8F cells through targeting VEGF

Document Type: Original Article

Authors

1 Department of Otorhinolaryngology, HwaMei Hospital, University of Chinese Academy of Sciences (Ningbo No.2 Hospital), Ningbo, Zhejiang 315010, P.R.China

2 Department of Clinical Laboratory, HwaMei Hospital, University of Chinese Academy of Sciences (Ningbo No.2 Hospital), Ningbo, Zhejiang 315010, P.R.China

Abstract

Objective(s): microRNA-29 (miR-29) family miRNAs have been mentioned as tumor suppressive genes in several human cancers. The purpose of this study was to investigate the function of miR-29a in nasopharyngeal carcinoma (NPC) cells.
Materials and Methods: Human NPC cell line 5-8F was transfected with mimic, inhibitor or scrambled controls specific for miR-29a. Subsequently, cell viability, migration, apoptosis and expression changes of VEGF were assessed by trypan blue staining, MTT assay, transwell assay, flow cytometry, Western blot and RT-qPCR. TargetScan online database was used to predict the targets of miR-29a, and luciferase reporter assay was carried out for testing the targeting relationship between VEGF and miR-29a. Western blot analysis was performed to determine the expression changes of core proteins in PI3K/AKT and JAK/STAT pathways.
Results: Overexpression of miR-29a suppressed 5-8F cells viability and relative migration, but increased apoptotic cell rate. Consistently, Bcl-2 was downregulated, Bax was upregulated, and caspase-3 and -9 were cleaved by miR-29a overexpression. VEGF was a target gene of miR-29a. Besides, VEGF silence exerted similar effects like miR-29a, as the viability and migration were repressed and apoptosis was induced. Finally, we found that PI3K/AKT and JAK/STAT pathways were deactivated by miR-29a or VEGF silence.
Conclusion: These findings highlighted the tumor suppressive effects of miR-29a on NPC cells, as its overexpression inhibited 5-8F cells viability, migration, and induced apoptosis. miR-29a exerted tumor suppressive functions might be via targeting VEGF and deactivating PI3K/AKT and JAK/STAT pathways.

Keywords

Main Subjects


1.Chang ET, Adami HO. The enigmatic epidemiology of nasopharyngeal carcinoma. Cancer Epidemiol Biomarkers Prev 2006; 15:1765-1777.

2.Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin 2015; 65:87-108.

3.Zhang F, Zhang J. Clinical hereditary characteristics in nasopharyngeal carcinoma through Ye-Liang's family cluster. Chin Med J 1999; 112:185-187.

4.Lee AW, Sze WM, Au JS, Leung SF, Leung TW, Chua DT, et al. Treatment results for nasopharyngeal carcinoma in the modern era: the Hong Kong experience. Int J Radiat Oncol Biol Phys 2005; 61:1107-1116.

5.Tan WL, Tan EH, Lim DW, Ng QS, Tan DS, Jain A, et al. Advances in systemic treatment for nasopharyngeal carcinoma. Chin Clin Oncol 2016; 5:21-32.

6.Feng X, Zhou S, Wang J, Hu W. MicroRNA profiles and functions in mosquitoes. PLoS Negl Trop Dis 2018; 12:1-19.

7.Rossi M, Amodio N, Di Martino MT, Tagliaferri P, Tassone P, Cho WC. MicroRNA and multiple myeloma: from laboratory findings to translational therapeutic approaches. Curr Pharm Biotechnol 2014; 15:459-467.

8.Rocci A, Hofmeister CC, Pichiorri F. The potential of miRNAs as biomarkers for multiple myeloma. Expert Rev Mol Diagn 2014; 14:947-959.

9.Wang CY, Ren JB, Liu M, Yu L. Targeting miR-29 induces apoptosis of osteosarcoma MG-63 cells via regulation of TGF-beta1/PUMA signal. Eur Rev Med Pharmacol Sci 2016; 20:3552-3560.

10.Zhu C, Wang Y, Kuai W, Sun X, Chen H, Hong Z. Prognostic value of miR-29a expression in pediatric acute myeloid leukemia. Clin Biochem 2013; 46:49-53.

11.Zhao X, Hou Y, Tuo Z, Wei F. Application values of miR-194 and miR-29 in the diagnosis and prognosis of gastric cancer. Exp Ther Med 2018; 15:4179-4184.

12.Al-Sheikh YA, Ghneim HK, Softa KI, Al-Jobran AA, Al-Obeed O, Mohamed MA, et al. Expression profiling of selected microRNA signatures in plasma and tissues of saudi colorectal cancer patients by qPCR. Oncol Lett 2016; 11:1406-1412.

13.Hong Q, Fang J, Pang Y, Zheng J. Prognostic value of the microRNA-29 family in patients with primary osteosarcomas. Med Oncol 2014; 31:37-45.

14.Park SY, Lee JH, Ha M, Nam JW, Kim VN. MiR-29 miRNAs activate p53 by targeting p85 alpha and CDC42. Nat Struct Mol Biol 2009; 16:23-29.

15.Xu H, Cheung IY, Guo HF, Cheung NK. MicroRNA miR-29 modulates expression of immunoinhibitory molecule B7-H3: potential implications for immune based therapy of human solid tumors. Cancer Res 2009; 69:6275-6281.

16.Wang HY, Yan LX, Shao Q, Fu S, Zhang ZC, Ye W, et al. Profiling plasma microRNA in nasopharyngeal carcinoma with deep sequencing. Clin Chem 2014; 60:773-782.

17.Gao J, Shao Z, Yan M, Fu T, Zhang L, Yan Y. Targetedregulationof STAT3 by miR-29a in mediating taxol resistance of nasopharyngeal carcinoma cell line CNE-1. Cancer Biomark 2018; 22:6441-648.

18.Qiu F, Sun R, Deng N, Guo T, Cao Y, Yu Y, et al. MiR-29a/b enhances cell migration and invasion in nasopharyngeal carcinoma progression by regulating SPARC and COL3A1 gene expression. PLoS One 2015; 10:1-22.

19.Chen L, Xiao H, Wang ZH, Huang Y, Liu ZP, Ren H, et al. MiR-29a suppresses growth and invasion of gastric cancer cells in vitro by targeting VEGF-A. BMB reports 2014; 47:39-44.

20.Chen J. Roles of the PI3K/Akt pathway in Epstein-Barr virus-induced cancers and therapeutic implications. World J Virol 2012; 1:154-161.

21.Pencik J, Pham HT, Schmoellerl J, Javaheri T, Schlederer M, Culig Z, et al. JAK-STAT signaling in cancer: From cytokines to non-coding genome. Cytokine 2016; 87:26-36.

22.Hu N.

23.Ko JY, Lee MS, Lian WS, Weng WT, Sun YC, Chen YS, et al. MicroRNA-29a counteracts synovitis in knee osteoarthritis pathogenesis by targeting VEGF. Sci Rep 2017; 7:3584-3598.

24.Meng H, Zhu X, Li L, Liang Z, Li X, Pan X, et al. Identification of CALM as the potential serum biomarker for predicting the recurrence of nasopharyngeal carcinoma using a mass spectrometry-based comparative proteomic approach. Int J Mol Med 2017; 40:1152-1164.

25.Deng B, Su F, Xie R, Tang W. MiR-371-5p suppresses the proliferative and migratory capacity of human nasopharyngeal carcinoma by targeting BCL2. Oncol Lett 2018; 15:9209-9215.

26.Jiang N, Jiang X, Chen Z, Song X, Wu L, Zong D, et al. MiR-203a-3p suppresses cell proliferation and metastasis through inhibiting LASP1 in nasopharyngeal carcinoma. J Exp Clin Cancer Res 2017; 36:138-148.

27.Qu R, Sun Y, Li Y, Hu C, Shi G, Tang Y, et al. MicroRNA-130a-3p suppresses cell viability, proliferation and invasion in nasopharyngeal carcinoma by inhibiting CXCL12. Am J Transl Res 2017; 9:3586-3598.

28.Luo Z, Zhang L, Li Z, Li X, Li G, Yu H, et al. An in silico analysis of dynamic changes in microRNA expression profiles in stepwise development of nasopharyngeal carcinoma. BMC Med Genomics 2012;5:3-15.

29.Zhang Y, Yang L, Wang S, Liu Z, Xiu M. MiR-29a suppresses cell proliferation by targeting SIRT1 in hepatocellular carcinoma. Cancer Biomark 2018; 22:151-159.

30.Liu X, Lv X, Yang Q, Jin H, Zhou W, Fan Q. MicroRNA-29a functions as a tumor suppressor and increases cisplatin sensitivity by targeting NRAS in lung cancer. Technol Cancer Res Treat 2018;17:1-8.

31.Shi C, Ren L, Sun C, Yu L, Bian X, Zhou X, et al. MiR-29a/b/c function as invasion suppressors for gliomas by targeting CDC42 and predict the prognosis of patients. Br J Cancer 2017; 117:1036-1047.

32.Li R, Liu J, Li Q, Chen G, Yu X. MiR-29a suppresses growth and metastasis in papillary thyroid carcinoma by targeting AKT3. Tumour Biol 2016; 37:3987-3996.

33.Liang C, Shi S, Meng Q, Liang D, Hua J, Qin Y, et al. MiR-29a, targeting caveolin 2 expression, is responsible for limitation of pancreatic cancer metastasis in patients with normal level of serum CA125. Int J Cancer 2018; 143:2919-2931.

34.Wang SC, Chai DS, Chen CB, Wang ZY, Wang L. HPIP promotes thyroid cancer cell growth, migration and EMT through activating PI3K/AKT signaling pathway. Biomed Pharmacother 2015; 75:33-39.

35.Slattery ML, Lundgreen A, Kadlubar SA, Bondurant KL, Wolff RK. JAK/STAT/SOCS-signaling pathway and colon and rectal cancer. Mol Carcinog 2013; 52:155-166.

36.Schmitt MJ, Philippidou D, Reinsbach SE, Margue C, Wienecke-Baldacchino A, Nashan D, et al. Interferon-gamma-induced activation of signal transducer and activator of transcription 1 (STAT1) up-regulates the tumor suppressing microRNA-29 family in melanoma cells. Cell Commun Signal 2012; 10:41-55.

37.Chen DD, Feng LC, Ye R, He YQ, Wang YD. MiR-29b reduces cisplatin resistance of gastric cancer cell by targeting PI3K/Akt Pathway. Zhongguo Yi Xue Ke Xue Yuan Xue Bao 2015; 37:514-519.