miR‑483‑5p promotes growth, invasion and self‑renewal of gastric cancer stem cells by Wnt/β‑catenin signaling

Wu,Kai; Ma,Longan; Zhu,Jinxiang

doi:10.3892/mmr.2016.5603

miR‑483‑5p promotes growth, invasion and self‑renewal of gastric cancer stem cells by Wnt/β‑catenin signaling

Authors:
- Kai Wu
- Longan Ma
- Jinxiang Zhu
View Affiliations

Affiliations: Department of General Surgery, The Third Affiliated Hospital (Shaanxi Provincial People's Hospital), Medical College of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China
Published online on: August 8, 2016 https://doi.org/10.3892/mmr.2016.5603
Pages: 3421-3428

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

Gastric carcinoma (GC) ranks as the second most common cause of cancer‑associated mortality worldwide. Emerging evidence has suggested a potential novel therapeutic strategy based on the ability of cancer stem cells (CSCs) to trigger tumorigenesis. MicroRNAs (miRNAs) have previously been implicated in CSC formation and regulation of their functional characteristics. In the current study, a significant upregulation of miR‑483‑5p levels was demonstrated in spheroid body‑forming cells (P<0.01) by reverse transcription‑quantitative polymerase chain reaction, which were isolated from the MKN‑45 gastric cancer cell line and possessed gastric CSC (GCSC) properties. An MTT assay demonstrated that overexpression of miR‑483‑5p by transfection with miR‑483‑5p mimics significantly increased cell proliferation and Annexin V‑propidium iodide staining indicated the suppression of cell apoptosis, suggesting that miR‑483‑5p has an important function in GCSC growth. Notably, Transwell and sphere formation assays demonstrated that miR‑483‑5p elevation promoted GCSC invasion and cell self‑renewal ability, respectively. Further western blotting assays demonstrated that miR‑483‑5p upregulation induced an increase in the protein expression levels of β‑catenin and its downstream target molecules, including cyclin D1, Bcl‑2 and matrix metalloproteinase 2, indicating that miR‑483‑5p activates Wnt/β‑catenin signaling. Inhibition of this pathway by β‑catenin small interfering RNA transfection attenuated the miR‑483‑5p‑induced effects on cell growth, invasion and self‑renewal. These results demonstrate that miR‑483‑5p may act as an oncogene to promote the development of GC by regulating GCSC growth, invasion and self‑renewal via the Wnt/β‑catenin signaling pathway. Thus, the present study suggests that miR‑483‑5p may be a promising therapeutic target against GC.

View Figures

View References

1	Gomceli I, Demiriz B and Tez M: Gastric carcinogenesis. World J Gastroenterol. 18:5164–5170. 2012.PubMed/NCBI
2	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
3	Brenner B, Hoshen MB, Purim O, David MB, Ashkenazi K, Marshak G, Kundel Y, Brenner R, Morgenstern S, Halpern M, et al: MicroRNAs as a potential prognostic factor in gastric cancer. World J Gastroenterol. 17:3976–3985. 2011. View Article : Google Scholar : PubMed/NCBI
4	Yu Z, Pestell TG, Lisanti MP and Pestell RG: Cancer stem cells. Int J Biochem Cell Biol. 44:2144–2151. 2012. View Article : Google Scholar : PubMed/NCBI
5	Xu G, Shen J, Ou Yang X, Sasahara M and Su X: Cancer stem cells: The 'heartbeat' of gastric cancer. J Gastroenterol. 48:781–797. 2013. View Article : Google Scholar
6	Visvader JE and Lindeman GJ: Cancer stem cells: Current status and evolving complexities. Cell Stem Cell. 10:717–728. 2012. View Article : Google Scholar : PubMed/NCBI
7	Conley SJ, Gheordunescu E, Kakarala P, Newman B, Korkaya H, Heath AN, Clouthier SG and Wicha MS: Antiangiogenic agents increase breast cancer stem cells via the generation of tumor hypoxia. Proc Natl Acad Sci USA. 109:2784–2789. 2012. View Article : Google Scholar : PubMed/NCBI
8	Yan K, Wu Q, Yan DH, Lee CH, Rahim N, Tritschler I, DeVecchio J, Kalady MF, Hjelmeland AB and Rich JN: Glioma cancer stem cells secrete Gremlin1 to promote their maintenance within the tumor hierarchy. Genes Dev. 28:1085–1100. 2014. View Article : Google Scholar : PubMed/NCBI
9	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: Accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar : PubMed/NCBI
10	Que T, Song Y, Liu Z, Zheng S, Long H, Li Z, Liu Y, Wang G, Liu Y, Zhou J, et al: Decreased miRNA-637 is an unfavorable prognosis marker and promotes glioma cell growth, migration and invasion via direct targeting Akt1. Oncogene. 34:4952–4963. 2015. View Article : Google Scholar : PubMed/NCBI
11	McGirt LY, Adams CM, Baerenwald DA, Zwerner JP, Zic JA and Eischen CM: miR-223 regulates cell growth and targets proto-oncogenes in mycosis fungoides/cutaneous T-cell lymphoma. J Invest Dermatol. 134:1101–1107. 2014. View Article : Google Scholar :
12	Chen PS, Su JL and Hung MC: Dysregulation of microRNAs in cancer. J Biomed Sci. 19:902012. View Article : Google Scholar : PubMed/NCBI
13	Croce CM and Calin GA: miRNAs, cancer, and stem cell division. Cell. 122:6–7. 2005. View Article : Google Scholar : PubMed/NCBI
14	Okada N, Lin CP, Ribeiro MC, Biton A, Lai G, He X, Bu P, Vogel H, Jablons DM, Keller AC, et al: A positive feedback between p53 and miR-34 miRNAs mediates tumor suppression. Genes Dev. 28:438–450. 2014. View Article : Google Scholar : PubMed/NCBI
15	Bao B, Wang Z, Ali S, Ahmad A, Azmi AS, Sarkar SH, Banerjee S, Kong D, Li Y, Thakur S and Sarkar FH: Metformin inhibits cell proliferation, migration and invasion by attenuating CSC function mediated by deregulating miRNAs in pancreatic cancer cells. Cancer Prev Res (Phila). 5:355–364. 2012. View Article : Google Scholar
16	Liu C, Kelnar K, Vlassov AV, Brown D, Wang J and Tang DG: Distinct microRNA expression profiles in prostate cancer stem/progenitor cells and tumor-suppressive functions of let-7. Cancer Res. 72:3393–3404. 2012. View Article : Google Scholar : PubMed/NCBI
17	Chabre O, Libé R, Assié G, Barreau O, Bertherat J, Bertagna X, Feige JJ and Cherradi N: Serum miR-483-5p and miR-195 are predictive of recurrence risk in adrenocortical cancer patients. Endocr Relat Cancer. 20:579–594. 2013.PubMed/NCBI
18	Zhang Z, Ge S, Wang X, Yuan Q, Yan Q, Ye H, Che Y, Lin Y, Zhang J and Liu P: Serum miR-483-5p as a potential biomarker to detect hepatocellular carcinoma. Hepatol Int. 7:199–207. 2013. View Article : Google Scholar : PubMed/NCBI
19	Song Q, Xu Y, Yang C, Chen Z, Jia C, Chen J, Zhang Y, Lai P, Fan X, Zhou X, et al: miR-483-5p promotes invasion and metastasis of lung adenocarcinoma by targeting RhoGDI1 and ALCAM. Cancer Res. 74:3031–3042. 2014. View Article : Google Scholar : PubMed/NCBI
20	Liu J, Ma L, Xu J, Liu C, Zhang J, Liu J, Chen R and Zhou Y: Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties. Int J Oncol. 42:453–459. 2013.
21	Wang L, Shi M, Hou S, Ding B, Liu L, Ji X, Zhang J and Deng Y: MiR-483-5p suppresses the proliferation of glioma cells via directly targeting ERK1. FEBS Lett. 586:1312–1317. 2012. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-timequantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
23	Wu H, Echt CS, Popp MP and Davis JM: Molecular cloning, structure and expression of an elicitor-inducible chitinase gene from pine trees. Plant Mol Biol. 33:979–987. 1997. View Article : Google Scholar : PubMed/NCBI
24	White BD, Chien AJ and Dawson DW: Dysregulation of Wnt/β-catenin signaling in gastrointestinal cancers. Gastroenterology. 142:219–232. 2012. View Article : Google Scholar
25	Yu T, Liu K, Wu Y, Fan J, Chen J, Li C, Yang Q and Wang Z: MicroRNA-9 inhibits the proliferation of oral squamous cell carcinoma cells by suppressing expression of CXCR4 via the Wnt/β-catenin signaling pathway. Oncogene. 33:5017–5027. 2014. View Article : Google Scholar
26	Brower JV, Clark PA, Lyon W and Kuo JS: MicroRNAs in cancer: Glioblastoma and glioblastoma cancer stem cells. Neurochem Int. 77:68–77. 2014. View Article : Google Scholar : PubMed/NCBI
27	Jiang J, Zhang Y, Chuai S, Wang Z, Zheng D, Xu F, Li C, Liang Y and Chen Z: Trastuzumab (herceptin) targets gastric cancer stem cells characterized by CD90 phenotype. Oncogene. 31:671–682. 2012. View Article : Google Scholar
28	Templeton AK, Miyamoto S, Babu A, Munshi A and Ramesh R: Cancer stem cells: Progress and challenges in lung cancer. Stem Cell Investigation. 1:92014.PubMed/NCBI
29	Qu X, Zhao M, Wu S, Yu W, Xu J, Xu J, Li J and Chen L: Circulating microRNA 483-5p as a novel biomarker for diagnosis survival prediction in multiple myeloma. Med Oncol. 31:2192014. View Article : Google Scholar : PubMed/NCBI
30	Arensman M, Lay AR, Kulikauskas RM, Chien AJ and Dawson DW: Wnt/β-catenin transcriptional activation promotes tumorigenesis and predicts survival in pancreatic cancer. Cancer Res. 73:40112013. View Article : Google Scholar
31	Holland JD, Klaus A, Garratt AN and Birchmeier W: Wnt signaling in stem and cancer stem cells. Curr Opin Cell Biol. 25:254–264. 2013. View Article : Google Scholar : PubMed/NCBI
32	Cai C and Zhu X: The Wnt/β-catenin pathway regulates self-renewal of cancer stem-like cells in human gastric cancer. Mol Med Rep. 5:1191–1196. 2012.PubMed/NCBI
33	Li J and Zhou BP: Activation of β-catenin and Akt pathways by Twist are critical for the maintenance of EMT associated cancer stem cell-like characters. BMC Cancer. 11:492011. View Article : Google Scholar
34	Kim HY, Park JH, Won HY, Lee JY and Kong G: CBX7 inhibits breast tumorigenicity through DKK-1-mediated suppression of the Wnt/β-catenin pathway. FASEB J. 29:300–313. 2015. View Article : Google Scholar