miR‑200b‑3p inhibits proliferation and induces apoptosis in colorectal cancer by targeting Wnt1

Chen,Lijuan; Wang,Xiangqun; Zhu,Yunhua; Zhu,Jian; Lai,Qingzhong

doi:10.3892/mmr.2018.9287

miR‑200b‑3p inhibits proliferation and induces apoptosis in colorectal cancer by targeting Wnt1

Authors:
- Lijuan Chen
- Xiangqun Wang
- Yunhua Zhu
- Jian Zhu
- Qingzhong Lai
View Affiliations

Affiliations: Department of Traditional Chinese Medicine, Yuhang Branch of Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, P.R. China, Department of Gastrointestinal Surgery, Yuhang Branch of Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, P.R. China, Massage Department, Hospital of Zhejiang Provincial Integrated Chinese and Western medicine, Hangzhou, Zhejiang 310003, P.R. China
Published online on: July 16, 2018 https://doi.org/10.3892/mmr.2018.9287
Pages: 2571-2580
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

MicroRNA (miR)‑200b‑3p is downregulated in multiple human cancer types. Wnt signaling serves a role in human colorectal cancer (CRC). The present study aimed to examine the effect of miR‑200b‑3p on human CRC and its potential association with Wnt signaling. The Cell Counting Kit‑8 (CCK‑8) was employed to assess cell viability. A flow cytometric assay was conducted to examine cell proliferation and apoptosis. The regulation model of miR‑200b‑3p and Wnt1 was assessed by a luciferase reporter assay. A commercial kit was used to evaluate the activity of caspase‑3 following treatment of the cells by miR‑200b‑3p or Wnt1. The expression of target factors was determined by a quantitative real‑time polymerase chain reaction and western blot analysis. The expression of miR‑200b‑3p was decreased in human CRC tissues and in cell lines. The bioinformatics analysis and the luciferase reporter assay revealed that Wnt1 may be a direct target of miR‑200b‑3p. Moreover, the viability and proliferation of CRC cells was suppressed by miR‑200b‑3p. miR‑200b‑3p additionally induced apoptosis in CRC cells. Furthermore, the caspase‑3 activity was enhanced in the miR‑200b‑3p mimics group. The expression of antigen Ki‑67 (additionally termed KI‑67) and β‑catenin was decreased, while the expression of cleaved caspase‑3 was increased by miR‑200b‑3p. In conclusion, miR‑200b‑3p inhibited proliferation and induced apoptosis in CRC cells by inactivating Wnt/β‑catenin signaling. The present study provided potential biomarkers and candidate modalities for the management of CRC.

View Figures

View References

1	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar : PubMed/NCBI
2	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2013. CA Cancer J Clin. 63:11–30. 2013. View Article : Google Scholar : PubMed/NCBI
3	Vaiopoulos AG, Athanasoula KC and Papavassiliou AG: Epigenetic modifications in colorectal cancer: Molecular insights and therapeutic challenges. Biochim Biophys Acta. 1842:971–980. 2014. View Article : Google Scholar : PubMed/NCBI
4	Watson AJM and Collins PD: Colon cancer: A civilization disorder. Dig Dis. 29:222–228. 2011. View Article : Google Scholar : PubMed/NCBI
5	Lee IM, Shiroma EJ, Lobelo F, Puska P, Blair SN and Katzmarzyk PT: Lancet Physical Activity Series Working Group: Effect of physical inactivity on major non-communicable diseases worldwide: An analysis of burden of disease and life expectancy. Lancet. 380:219–229. 2012. View Article : Google Scholar : PubMed/NCBI
6	Fearon ER, Pardoll DM, Itaya T, Golumbek P, Levitsky HI, Simons JW, Karasuyama H, Vogelstein B and Frost P: Interleukin-2 production by tumor cells bypasses T helper function in the generation of an antitumor response. Cell. 60:397–403. 1990. View Article : Google Scholar : PubMed/NCBI
7	Basu S, Haase G and Ben-Ze'ev A: Wnt signaling in cancer stem cells and colon cancer metastasis. F1000Res. 19:52016.
8	Giles RH, van Es JH and Clevers H: Caught up in a wnt storm: Wnt signaling in cancer. Biochim Biophys Acta. 1653:1–24. 2003.PubMed/NCBI
9	Capdevila J and Belmonte lzpisúa JC: Extracellular modulation of the Hedgehog, Wnt and TGF-beta signalling pathways during embryonic development. Curr Opin Genet Dev. 9:427–433. 1999. View Article : Google Scholar : PubMed/NCBI
10	Tanaka S, Terada K and Nohno T: Canonical Wnt signaling is involved in switching from cell proliferation to myogenic differentiation of mouse myoblast cells. J Mol Signal. 6:122011. View Article : Google Scholar : PubMed/NCBI
11	Chen S, Guttridge DC, You Z, Zhang Z, Fribley A, Mayo MW, Kitajewski J and Wang CY: WNT-1 Signaling Inhibits Apoptosis by Activating β-Catenin/T Cell Factor–Mediated Transcription. J Cell Biol. 152:87–96. 2001. View Article : Google Scholar : PubMed/NCBI
12	Logan CY and Nusse R: The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol. 20:781–810. 2004. View Article : Google Scholar : PubMed/NCBI
13	Cadigan KM and Nusse R: Wnt signaling: A common theme in animal development. Genes Dev. 11:3286–3305. 1997. View Article : Google Scholar : PubMed/NCBI
14	Dai X, Wang L, Zhang L, Han Y, Yang G and Li L: The expression and mutation of beta-catenin in colorectal traditional serrated adenomas. Indian J Pathol Microbiol. 55:288–293. 2012. View Article : Google Scholar : PubMed/NCBI
15	Pasquinelli AE, Reinhart BJ, Slack F, Martindale MQ, Kuroda MI, Maller B, Hayward DC, Ball EE, Degnan B, Muller P, et al: Conservation of the sequence and temporal expression of let-7 heterochronic regulatory RNA. Nature. 408:86–89. 2000. View Article : Google Scholar : PubMed/NCBI
16	Wang XJ, Reyes JL, Chua NH and Gaasterland T: Prediction and identification of Arabidopsis thaliana microRNAs and their mRNA targets. Genome Biol. 5:E652004. View Article : Google Scholar
17	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
18	Michael MZ, SM OC, van Holst Pellekaan NG, Young GP and James RJ: Reduced accumulation of specific microRNAs in colorectal neoplasia. Mol Cancer Res. 1:882–891. 2003.PubMed/NCBI
19	Fan D, Lin X, Zhang F, Zhong W, Hu J, Chen Y, Cai Z, Zou Y, He X, Chen X, et al: MicroRNA 26b promotes colorectal cancer metastasis by down-regulating pten and wnt5a. Cancer Sci. 109:354–362. 2017. View Article : Google Scholar : PubMed/NCBI
20	Hashimi ST, Fulcher JA, Chang MH, Gov L, Wang S and Lee B: MicroRNA profiling identifies miR-34a and miR-21 and their target genes JAG1 and WNT1 in the coordinate regulation of dendritic cell differentiation. Blood. 114:404–414. 2009. View Article : Google Scholar : PubMed/NCBI
21	Huang S, Zhang L, Yang P, Chen P and Xie Y: HCV core protein-induced down-regulation of microRNA-152 promoted aberrant proliferation by targeting Wnt1 in HepG2 cells. PLOS One. 9:e817302014. View Article : Google Scholar : PubMed/NCBI
22	Si W, Li Y, Shao H, Hu R, Wang W, Zhang K and Yang Q: MiR-34a Inhibits Breast Cancer Proliferation and Progression by Targeting Wnt1 in Wnt/β-catenin Signaling Pathway. Am J Med Sci. 352:191–199. 2016. View Article : Google Scholar : PubMed/NCBI
23	Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y and Goodall GJ: The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 10:593–601. 2008. View Article : Google Scholar : PubMed/NCBI
24	Kinzler KW and Vogelstein B: Lessons from hereditary colorectal cancer. Cell. 87:159–170. 1996. View Article : Google Scholar : PubMed/NCBI
25	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
26	Williams LV, Veliceasa D, Vinokour E and Volpert OV: miR-200b inhibits prostate cancer EMT, growth and metastasis. PLoS One. 8:e839912013. View Article : Google Scholar : PubMed/NCBI
27	Wei W, Chua MS, Grepper S and So SK: Blockade of Wnt-1 signaling leads to anti-tumor effects in hepatocellular carcinoma cells. Mol Cancer. 8:762009. View Article : Google Scholar : PubMed/NCBI
28	Carthew RW and Sontheimer EJ: Origins and Mechanisms of miRNAs and siRNAs. Cell. 136:642–655. 2009. View Article : Google Scholar : PubMed/NCBI
29	Porter AG and Janicke RU: Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 6:99–104. 1999. View Article : Google Scholar : PubMed/NCBI
30	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
31	Gao ZH, Lu C, Wang MX, Han Y and Guo LJ: Differential β-catenin expression levels are associated with morphological features and prognosis of colorectal cancer. Oncol Lett. 8:2069–2076. 2014. View Article : Google Scholar : PubMed/NCBI
32	Leskela S, Leandro-Garcia LJ, Mendiola M, Barriuso J, Inglada-Perez L, Munoz I, Martinez-Delgado B, Redondo A, de Santiago J, Robledo M, et al: The miR-200 family controls beta-tubulin III expression and is associated with paclitaxel-based treatment response and progression-free survival in ovarian cancer patients. Endocr Relat Cancer. 18:85–95. 2010. View Article : Google Scholar : PubMed/NCBI
33	Yao Y, Hu J, Shen Z, Yao R, Liu S, Li Y, Cong H, Wang X, Qiu W and Yue L: MiR-200b expression in breast cancer: A prognostic marker and act on cell proliferation and apoptosis by targeting Sp1. J Cell Mol Med. 19:760–769. 2015. View Article : Google Scholar : PubMed/NCBI
34	He B, You L, Uematsu K, Xu Z, Lee AY, Matsangou M, Mccormick F and Jablons DM: A monoclonal antibody against Wnt-1 induces apoptosis in human cancer cells. Neoplasia. 6:7–14. 2004. View Article : Google Scholar : PubMed/NCBI
35	Humphries B and Yang C: The microRNA-200 family: Small molecules with novel roles in cancer development, progression and therapy. Oncotarget. 6:6472–6498. 2015. View Article : Google Scholar : PubMed/NCBI
36	Ghavami S, Hashemi M, Ande SR, Yeganeh B, Xiao W, Eshraghi M, Bus CJ, Kadkhoda K, Wiechec E, Halayko AJ and Los M: Apoptosis and cancer: Mutations within caspase genes. J Med Genet. 46:497–510. 2009. View Article : Google Scholar : PubMed/NCBI
37	Boatright KM and Salvesen GS: Mechanisms of caspase activation. Curr Opin Cell Biol. 15:725–731. 2003. View Article : Google Scholar : PubMed/NCBI
38	Bruno S and Darzynkiewicz Z: Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60 cells. Cell Prolif. 25:31–40. 1992. View Article : Google Scholar : PubMed/NCBI
39	Rafael S, Veganzones S, Vidaurreta M, de la Orden V and Maestro ML: Effect of β-catenin alterations in the prognosis of patients with sporadic colorectal cancer. J Cancer Res Ther. 10:591–596. 2014.PubMed/NCBI
40	Jung YS, Jun S, Lee SH, Sharma A and Park JI: Wnt2 complements Wnt/β-catenin signaling in colorectal cancer. Oncotarget. 6:37257–37268. 2015. View Article : Google Scholar : PubMed/NCBI
41	Clevers H: Wnt/beta-catenin signaling in development and disease. Cell. 127:469–480. 2006. View Article : Google Scholar : PubMed/NCBI
42	Chen DQ, Pan BZ, Huang JY, Zhang K, Cui SY, De W, Wang R and Chen LB: HDAC 1/4-mediated silencing of microRNA-200b promotes chemoresistance in human lung adenocarcinoma cells. Oncotarget. 5:3333–3349. 2014. View Article : Google Scholar : PubMed/NCBI
43	Kolesnikoff N, Attema JL, Roslan S, Bert AG, Schwarz QP, Gregory PA and Goodall GJ: Specificity protein 1 (Sp1) maintains basal epithelial expression of the miR-200 family: Implications for epithelial-mesenchymal transition. J Biol Chem. 289:11194–11205. 2014. View Article : Google Scholar : PubMed/NCBI
44	Manavalan TT, Teng Y, Litchfield LM, Muluhngwi P, Al-Rayyan N and Klinge CM: Reduced expression of miR-200 family members contributes to antiestrogen resistance in LY2 human breast cancer cells. PLOS One. 8:e623342013. View Article : Google Scholar : PubMed/NCBI