MicroRNA‑944 targets vascular endothelial growth factor to inhibit cell proliferation and invasion in osteosarcoma

Yan,Tingzhen; Zhu,Shiyong; Zhang,Jing; Lu,Gongbiao; Lv,Chaoliang; Wei,Yanchun; Luo,Minghua

doi:10.3892/mmr.2018.9524

MicroRNA‑944 targets vascular endothelial growth factor to inhibit cell proliferation and invasion in osteosarcoma

Authors:
- Tingzhen Yan
- Shiyong Zhu
- Jing Zhang
- Gongbiao Lu
- Chaoliang Lv
- Yanchun Wei
- Minghua Luo
View Affiliations

Affiliations: Department of Spine Surgery, Jining No. 1 People's Hospital, Jining, Shandong 272011, P.R. China, Department of Spine Surgery, Lanling County People's Hospital of Shandong Province, Linyi, Shandong 277700, P.R. China, Department of Pathology, Jining No. 1 People's Hospital, Jining, Shandong 272011, P.R. China, Department of Orthopedics, The Second People's Hospital of Jingdezhen, Jingdezhen, Jiangxi 333000, P.R. China
Published online on: October 1, 2018 https://doi.org/10.3892/mmr.2018.9524
Pages: 5221-5228

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

Emerging evidence has demonstrated that the dysregulation of microRNA (miRNA/miR) serves a crucial role in the tumorigenesis and tumor development of osteosarcoma (OS), primarily by affecting various pathological behaviors. Therefore, better knowledge of miRNA in OS may provide novel insights into the pathogenesis of OS, and may facilitate the development of promising therapeutics for patients with this disease. MiRNA‑944 is frequently dysregulated in human cancers. However, the expression levels, functions and underlying mechanisms of miR‑944 in OS remain largely elusive. In the present study, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was performed to detect miR‑944 expression in OS tissues and cell lines. The regulatory influence of miR‑944 in OS proliferation and invasion was determined with MTT and Transwell invasion assays. In addition, the mechanisms underlying the action of miR‑944 in OS cells were elucidated through a series of experiments, including bioinformatics analysis, luciferase reporter assay, RT‑qPCR and western blot analysis. Spearman's correlation analysis was utilized to examine the relationship between miR‑944 and VEGF expression levels, and rescue experiments were applied to further verify whether VEGF mediates the role of miR‑944 in OS. The results demonstrated that miR‑944 was downregulated in cancer tissues and cell lines. Furthermore, exogenous miR‑944 expression inhibited cell proliferation and invasion in OS in vitro. Vascular endothelial growth factor (VEGF) was identified as a direct target of miR‑944 in OS and was overexpressed in cancer tissues. VEGF expression was inversely correlated with miR‑944 expression in cancer tissues. Rescue experiments demonstrated that overexpression of VEGF partially prevented the miR‑944‑induced inhibition of OS cell proliferation and invasion. These results suggested that miR‑944 may serve a tumor suppressive role in OS by directly targeting VEGF. Therefore, miR‑944 may be a promising target in the treatment of OS.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 66:7–30. 2016. View Article : Google Scholar : PubMed/NCBI
2	Kansara M, Teng MW, Smyth MJ and Thomas DM: Translational biology of osteosarcoma. Nat Rev Cancer. 14:722–735. 2014. View Article : Google Scholar : PubMed/NCBI
3	Gianferante DM, Mirabello L and Savage SA: Germline and somatic genetics of osteosarcoma-connecting aetiology, biology and therapy. Nat Rev Endocrinol. 13:480–491. 2017. View Article : Google Scholar : PubMed/NCBI
4	Fagioli F, Aglietta M, Tienghi A, Ferrari S, del Prever Brach A, Vassallo E, Palmero A, Biasin E, Bacci G, Picci P and Madon E: High-dose chemotherapy in the treatment of relapsed osteosarcoma: An Italian sarcoma group study. J Clin Oncol. 20:2150–2156. 2002. View Article : Google Scholar : PubMed/NCBI
5	Wada T, Isu K, Takeda N, Usui M, Ishii S and Yamawaki S: A preliminary report of neoadjuvant chemotherapy NSH-7 study in osteosarcoma: Preoperative salvage chemotherapy based on clinical tumor response and the use of granulocyte colony-stimulating factor. Oncology. 53:221–227. 1996. View Article : Google Scholar : PubMed/NCBI
6	Tan ML, Choong PF and Dass CR: Osteosarcoma: Conventional treatment vs. gene therapy. Cancer Biol Ther. 8:106–117. 2009. View Article : Google Scholar : PubMed/NCBI
7	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. 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	Lewis BP, Burge CB and Bartel DP: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 120:15–20. 2005. View Article : Google Scholar : PubMed/NCBI
10	Lin S and Gregory RI: MicroRNA biogenesis pathways in cancer. Nat Rev Cancer. 15:321–333. 2015. View Article : Google Scholar : PubMed/NCBI
11	Ghosh T, Varshney A, Kumar P, Kaur M, Kumar V, Shekhar R, Devi R, Priyanka P, Khan MM and Saxena S: MicroRNA-874-mediated inhibition of the major G₁/S phase cyclin, CCNE1, is lost in osteosarcomas. J Biol Chem. 292:21264–21281. 2017. View Article : Google Scholar : PubMed/NCBI
12	Li D, Wang H, Song H, Xu H, Zhao B, Wu C, Hu J, Wu T, Xie D, Zhao J, et al: The microRNAs miR-200b-3p and miR-429-5p target the LIMK1/CFL1 pathway to inhibit growth and motility of breast cancer cells. Oncotarget. 8:85276–85289. 2017.PubMed/NCBI
13	Huang RS, Zheng YL, Zhao J and Chun X: microRNA-381 suppresses the growth and increases cisplatin sensitivity in non-small cell lung cancer cells through inhibition of nuclear factor-kB signaling. Biomed Pharmacother. 98:538–544. 2018. View Article : Google Scholar : PubMed/NCBI
14	Zhao F, Pu Y, Qian L, Zang C, Tao Z and Gao J: MiR-20a-5p promotes radio-resistance by targeting NPAS2 in nasopharyngeal cancer cells. Oncotarget. 8:105873–105881. 2017. View Article : Google Scholar : PubMed/NCBI
15	Zhao Y, Liu X and Lu YX: MicroRNA-143 regulates the proliferation and apoptosis of cervical cancer cells by targeting HIF-1α. Eur Rev Med Pharmacol Sci. 21:5580–5586. 2017.PubMed/NCBI
16	He Y and Yu B: MicroRNA-93 promotes cell proliferation by directly targeting P21 in osteosarcoma cells. Exp Ther Med. 13:2003–2011. 2017. View Article : Google Scholar : PubMed/NCBI
17	Lin W, Zhu X, Yang S, Chen X, Wang L, Huang Z, Ding Y, Huang L and Lv C: MicroRNA-203 inhibits proliferation and invasion, and promotes apoptosis of osteosarcoma cells by targeting Runt-related transcription factor 2. Biomed Pharmacother. 91:1075–1084. 2017. View Article : Google Scholar : PubMed/NCBI
18	He Z, Xu H, Meng Y and Kuang Y: miR-944 acts as a prognostic marker and promotes the tumor progression in endometrial cancer. Biomed Pharmacother. 88:902–910. 2017. View Article : Google Scholar : PubMed/NCBI
19	Xie H, Lee L, Scicluna P, Kavak E, Larsson C, Sandberg R and Lui WO: Novel functions and targets of miR-944 in human cervical cancer cells. Int J Cancer. 136:E230–E241. 2015. View Article : Google Scholar : PubMed/NCBI
20	He H, Tian W, Chen H and Jiang K: MiR-944 functions as a novel oncogene and regulates the chemoresistance in breast cancer. Tumour Biol. 37:1599–1607. 2016. View Article : Google Scholar : PubMed/NCBI
21	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
22	Liu Y, Zhang F, Zhang Z, Wang D, Cui B, Zeng F, Huang L, Zhang Q and Sun Q: High expression levels of Cyr61 and VEGF are associated with poor prognosis in osteosarcoma. Pathol Res Pract. 213:895–899. 2017. View Article : Google Scholar : PubMed/NCBI
23	Yu XW, Wu TY, Yi X, Ren WP, Zhou ZB, Sun YQ and Zhang CQ: Prognostic significance of VEGF expression in osteosarcoma: A meta-analysis. Tumour Biol. 35:155–160. 2014. View Article : Google Scholar : PubMed/NCBI
24	Mei J, Gao Y, Zhang L, Cai X, Qian Z, Huang H and Huang W: VEGF-siRNA silencing induces apoptosis, inhibits proliferation and suppresses vasculogenic mimicry in osteosarcoma in vitro. Exp Oncol. 30:29–34. 2008.PubMed/NCBI
25	Peng N, Gao S, Guo X, Wang G, Cheng C, Li M and Liu K: Silencing of VEGF inhibits human osteosarcoma angiogenesis and promotes cell apoptosis via VEGF/PI3K/AKT signaling pathway. Am J Transl Res. 8:1005–1015. 2016.PubMed/NCBI
26	Kushlinskii NE, Fridman MV and Braga EA: Molecular mechanisms and microRNAs in osteosarcoma pathogenesis. Biochemistry (Mosc). 81:315–328. 2016. View Article : Google Scholar : PubMed/NCBI
27	Sampson VB, Yoo S, Kumar A, Vetter NS and Kolb EA: MicroRNAs and potential targets in osteosarcoma: Review. Front Pediatr. 3:692015. View Article : Google Scholar : PubMed/NCBI
28	Wen L, Li Y, Jiang Z, Zhang Y, Yang B and Han F: miR-944 inhibits cell migration and invasion by targeting MACC1 in colorectal cancer. Oncol Rep. 37:3415–3422. 2017. View Article : Google Scholar : PubMed/NCBI
29	Pan T, Chen W, Yuan X, Shen J, Qin C and Wang L: miR-944 inhibits metastasis of gastric cancer by preventing the epithelial-mesenchymal transition via MACC1/Met/AKT signaling. FEBS Open Bio. 7:905–914. 2017. View Article : Google Scholar : PubMed/NCBI
30	Liu M, Zhou K and Cao Y: MicroRNA-944 affects cell growth by targeting EPHA7 in non-small cell lung cancer. Int J Mol Sci. 17:pii: E1493. 2016. View Article : Google Scholar
31	Ji YN, Wang Q, Li Y and Wang Z: Prognostic value of vascular endothelial growth factor A expression in gastric cancer: A meta-analysis. Tumour Biol. 35:2787–2793. 2014. View Article : Google Scholar : PubMed/NCBI
32	Tian XF, Zhang XW, Chen RX, Wang JW and Zhang D: Clinical significance of expression of VEGF and bFGF in thyroid carcinoma. Zhonghua Wai Ke Za Zhi. 42:864–866. 2004.(In Chinese). PubMed/NCBI
33	Huang YJ, Qi WX, He AN, Sun YJ, Shen Z and Yao Y: Prognostic value of tissue vascular endothelial growth factor expression in bladder cancer: A meta-analysis. Asian Pac J Cancer Prev. 14:645–649. 2013. View Article : Google Scholar : PubMed/NCBI
34	Wen L, Wang R, Lu X and You C: Expression and clinical significance of vascular endothelial growth factor and fms-related tyrosine kinase 1 in colorectal cancer. Oncol Lett. 9:2414–2418. 2015. View Article : Google Scholar : PubMed/NCBI