The effect of baicalein on Wnt/ß-catenin pathway and miR-25 expression in Saos-2 osteosarcoma cell line

Background/aim: Osteosarcoma is the most common primary bone malignancy that occurs frequently in children and adolescents. Baicalein, a flavonoid that has attracted great attention in recent years with its strong antitumor activity, shows a wide range of biological and pharmaceutical effects.MicroRNAs have been found to be involved in many critical processes in cancers. This study aimed to investigate the effect of baicalein and miR-25 on Wnt/ß-catenin signaling pathway of osteosarcoma cell line Saos-2. Materials and methods: Cell viability was assessed, and qRT-PCR and Western blot were performed to study the effects of baicalein on expression of Wnt/ß-catenin signaling pathway-realted genes ß-catenin, GSK-3ß, and Axin2 of Saos-2 cells. Results: Our results indicated that baicalein can inhibit the proliferation IC50 value 35 -M , regulate Wnt/ß-catenin pathway and also increase miR-25 expression of Saos-2. Baicalein and also miR-25 decreased the expression of ß-catenin and Axin2, while increasing the expression of GSK-3ß. Down regulation of miR-25 decreased the expression of GSK-3ß, while ß-catenin and Axin2 expression increased. Conclusion: These findings demonstrate that baicalein may target genes related to the Wnt/ß-catenin pathway by regulating miR-25 expression and may be a potential Wnt/ß-catenin pathway inhibitor for osteosarcoma therapy.

___

  • 1. Broadhead ML, Clark JCM, Myers D, Dass CR, Choong PFM. The molecular pathogenesis of osteosarcoma: a review. Sarcoma 2011; 1-12. doi: 10.1155/2011/959248
  • 2. Amado NG, Fonseca BF, Cerqueira DM, Neto VM, Abreu JG. Flavonoids: potential Wnt/β-catenin signaling modulators in cancer. Life Sciences 2011; 89: 545-554. doi: 10.1016/j. lfs.2011.05.003
  • 3. Park S, Choi J. Inhibition of β-catenin/Tcf signaling by flavonoids. Journal of Cellular Biochemistry 2010; 110: 1376- 1385. doi: 10.1002/jcb.22654
  • 4. Chen J, Li Z, Chen AY, Ye X, Luo H et al. Inhibitory effect of baicalin and baicalein on ovarian cancer cells. International Journal of Molecular Sciences 2013; 14 (3): 6012-6025. doi: 10.3390/ijms14036012
  • 5. Ye F, Wang H, Zhang L, Zou Y, Han H et al. Baicalein induces human osteosarcoma cell line MG-63 apoptosis via ROSinduced BNIP3 expression. Tumor Biology 2015; 36: 4731- 4740. doi: 10.1007/s13277-015-3122-y
  • 6. Guo Z, Hu X, Xing Z, Xing R, Lv R et al. Baicalein inhibits prostate cancer cell growth and metastasis via the caveolin-1/ AKT/mTOR pathway. Molecular and Cellular Biochemistry 2015; 406: 111-119. doi: 10.1007/s11010-015-2429-8
  • 7. Li HL, Zhang S, Wang Y, Liang RR, Li J et al. Baicalein induces apoptosis via a mitochondrial-dependent caspase activation pathway in T24 bladder cancer cells. Molecular Medicine Reports 2013; 7: 266-270. doi: 10.3892/mmr.2012.1123
  • 8. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116: 281-297. doi: 10.1016/s0092- 8674(04)00045-5
  • 9. Miao J, Wu S, Peng Z, Tania M, Zhang C. MicroRNAs in osteosarcoma: diagnostic and therapeutic aspects. Tumor Biology 2013; 34: 2093-2098. doi: 10.1007/s13277-013-0940-7
  • 10. Kumar RMR, Boro A, Fuchs B. Involvement and clinical aspects of MicroRNA in osteosarcoma. International Journal of Molecular Sciences 2016; 17: 877. doi: 10.3390/ijms17060877
  • 11. Xu Z, Li C, Qu H, Li H, Gu Q et al. MicroRNA-195 inhibits the proliferation and invasion of pancreatic cancer cells by targeting the fatty acid synthase/Wnt signaling pathway. Tumour Biology 2017; 39 (6): 1010428317711324. doi: 10.1177/1010428317711324
  • 12. Li H, Zhang K, Liu LH, Ouyang Y, Guo HB et al. MicroRNA screening identifies circulating microRNAs as potential biomarkers for osteosarcoma. Oncology Letters 2015; 10: 1662-1668. doi: 10.3892/ol.2015.3378
  • 13. Zhu S, Pan W, Qian Y. MicroRNA in immunity and autoimmunity. Journal of Molecular Medicine 2013; 91: 1039- 1050. doi: 10.1007/s00109-013-1043-z
  • 14. Bian S. and Sun T. Functions of noncoding RNAs in neural development and neurological diseases. Molecular Neurobiology 2011; 44 (3): 359-373. doi: 10.1007/s12035-011- 8211-3
  • 15. Guo Y and Niu S.MiR-25 protects PC-12 cells from H2O2 mediated oxidative damage via WNT/β-catenin pathway. The Journal of Spinal Cord Medicine 2017; 12: 1-10. doi: 10.1080/10790268.2017.1336319
  • 16. Kocyigit I, Taheri S, Sener EF, Eroglu E, Ozturk F et al. Serum micro-RNA profiles in patients with autosomal dominant polycystic kidney disease according to hypertension and renal function. BMC Nephrology 2017; 18: 179. doi: 10.1186/s12882- 017-0600-z
  • 17. Huo J, Zhang Y, Li R, Wang Y, Wu J et al.Upregulated MicroRNA-25 mediates the migration of melanoma cells by targeting DKK3 through the WNT/β-Catenin pathway. International Journal of Molecular Sciences 2016; 17: 1124. doi: 10.3390/ijms17111124
  • 18. Li Q, ZouC, Zou C, Han Z, Xiao H et al. MicroRNA-25 functions as a potential tumor suppressor in colon cancer by targeting Smad7. Cancer Letters 2013; 335: 168-174. doi: 10.1016/j.canlet.2013.02.029
  • 19. Kim YK, Yu J, Han TS, Park SY, Namkoong B et al. Functional links between clustered microRNAs: suppression of cell-cycle inhibitors by microRNA clusters in gastric cancer. Nucleic Acids Research 2009; 37: 1672-1681. doi: 10.1093/nar/gkp002
  • 20. Poliseno L, Salmena L, Riccardi L, Fornari A, SongMS et al. Identification of the miR-106b~25 microRNA cluster as a proto-oncogenic PTEN-targeting intron that cooperates with its host gene MCM7 in transformation. Science Signaling 2010; 3 (117): ra29. doi: 10.1126/scisignal.2000594
  • 21. Feng X, Jiang J, Shi S, Xie H, Zhou L et al. Knockdown of miR25 increases the sensitivity of liver cancer stem cells to TRAILinduced apoptosis via PTEN/PI3K/Akt/Bad signaling pathway. International Journal of Oncology 2016; 49: 2600-2610. doi: 10.3892/ijo.2016.3751
  • 22. He N, Zhang Z. Baicalein suppresses the viability of MG-63 osteosarcoma cells through inhibiting c-MYC expression via Wnt signaling pathway. Molecular and Cellular Biochemistry 2015; 405: 187-196. doi: 10.1007/s11010-015-2410-6
  • 23. Liu B, Li G, Wang X, Liu Y. A furin inhibitor downregulates osteosarcoma cell migration by downregulating the expression levels of MT1 MMP via the Wnt signaling pathway. Oncology Letters 2014; 7: 1033-1038. doi: 10.3892/ol.2014.1839
  • 24. Leichter AL, Sullivan MJ, Eccles MR, Chatterjee A. MicroRNA expression patterns and signalling pathways in the development and progression of childhood solid tumours. Molecular Cancer 2017; 16: 15. doi: 10.1186/s12943-017-0584-0
  • 25. Anton R, Chatterjee SS, Simundza J, Cowin P, Dasgupta R. A systematic screen for Micro-RNAs regulating the canonical Wnt pathway. PLoS One 2011; 6 (10): e26257. doi: 10.1371/ journal.pone.0026257
  • 26. Ma Y, Zhu B, Liu X, Yu H, Yong L et al. Inhibition of oleandrin on the proliferation show and invasion of osteosarcoma cells in vitro by suppressing Wnt/β-catenin signaling pathway. Journal of Experimental & Clinical Cancer Research 2015; 34: 115. doi: 10.1186/s13046-015-0232-8
  • 27. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta DeltaC(T)) method. Methods 2001; 25 (4): 402-408. doi: 10.1006/meth.2001.1262
  • 28. Han Z, Zhu S, Han X, Wang Z, Wu S et al. Baicalein inhibits hepatocellular carcinoma cells through suppressing the expression of CD24. International Immunopharmacology 2015; 29: 416-422. doi: 10.1016/j.intimp.2015.10.021
  • 29. Liu X, Liu S, Chen J, He L, Meng X et al. Baicalein suppresses the proliferation of acute T-lymphoblastic leukemia Jurkat cells by inhibiting the Wnt/β-catenin signaling. Annals of Hematology 2016; 95: 1787-1793. doi: 10.1007/s00277-016-2766-z
  • 30. Vega OA, Lucero CMJ, Araya HF, Jerez S, Tapia JC et al. Wnt/ β-catenin signaling activates expression of the bone-related transcription factor RUNX2 in select human osteosarcoma cell types. Journal of Cellular Biochemistry 2017; 118: 3662-3674. doi: 10.1002/jcb.26011
  • 31. Haydon RC, Deyrup A, Ishikawa A, Heck R, Jiang W et al.Cytoplasmic and/or nuclear accumulation of the β-catenin protein is a frequent event in human osteosarcoma. International Journal of Cancer 2002; 102: 338-342. doi: 10.1002/ijc.10719
  • 32. Huang K, Zhang J, Han L, You Y, Jiang T et al. MicroRNA roles in beta-catenin pathway. Molecular Cancer 2010; 9: 252-263. doi: 10.1186/1476-4598-9-252
  • 33. Stolz A, Bastians H. Fresh WNT into the regulation of mitosis. Cell Cycle 2015; 14: 2566-2570. doi: 10.1080/15384101.2015.1064569
  • 34. Taketo MM. Shutting down Wnt signal-activated cancer. Nature Genetics 2004; 36: 320-322. doi: 10.1038/ng0404-320
  • 35. Lustig B, Jerchow B, Sachs M, Weiler S, Pietsch T et al. Negative feedback loop of Wnt signaling through upregulation of conductin/axin2 in colorectal and liver tumors. Molecular and Cellular Biology 2002; 22: 1184-1193. doi: 10.1128/ mcb.22.4.1184-1193.2002
  • 36. Onyido EK, Sweeney E, Nateri AS. Wnt-signalling pathways and microRNAs network in carcinogenesis: experimental and bioinformatics approaches. Molecular Cancer 2016; 15: 56. doi: 10.1186/s12943-016-0541-3
  • 37. Ma X, Yan W, Dai Z, Gao X, Ma Y et al. Baicalein suppresses metastasis of breast cancer cells by inhibiting EMT via downregulation of SATB1 and Wnt/β-catenin pathway. Drug Design, Development and Therapy 2016;10: 1419-1441. doi: 10.2147/DDDT.S102541
  • 38. Song JL, Nigam P, Tektas SS, Selva E. MicroRNA regulation of Wnt signaling pathways in development and disease. Cell Signalling 2015; 27: 1380-1391. doi: 10.1016/j.cellsig.2015.03.018
  • 39. Suzuki A, Ozono K, Kubota T, Kondou H, Tachikawa K et al. PTH/cAMP/PKA signaling facilitates canonical Wnt signaling via inactivation of glycogen synthase kinase-3β in osteoblastic Saos-2 Cells. Journal of Cellular Biochemistry 2008; 104: 304- 317. doi: 10.1002/jcb.21626
  • 40. Yan K, Gao J, Yang T, Ma Q, Qiu X et al. MicroRNA-34a inhibits the proliferation and metastasis of osteosarcoma cells both in vitro and in vivo. PLoS One 2012; 7 (3): e33778. doi: 10.1371/ journal.pone.0033778
Turkish Journal of Medical Sciences-Cover
  • ISSN: 1300-0144
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Ömer KUTLU, Zeynal DOĞAN, Hatice Meral EKŞİOĞLU, Murat KEKİLLİ

Evaluation of the cardioprotective effects of crystalloid del Nido cardioplegia solution via a rapid and accurate cardiac marker: heart-type fatty acid-binding protein

Duygun ALTINTAŞ AYKAN, Aydemir KOÇARSLAN, Adem DOĞANER, Yavuz ORAK, Mehmet KİRİŞCİ, Filiz ALKAN BAYLAN

Subclinical cardiac structural and electrical abnormalities in fibromyalgia syndrome

Ekrem AKSU, Abdullah SÖKMEN, Gülizar SÖKMEN, Enes ÇELİK, Ejder BERK

Cross-cultural adaptation, reliability and validity of the Turkish version of Patient-Specific Functional Scale in patients with chronic neck pain

Gamze YALÇINKAYA, Bilge KARA, Mehmet Nuri ARDA

Ekrem AKSU, Ejder BERK, Abdullah SÖKMEN, Gülizar SÖKMEN, Enes ÇELİK

Should the dose of levothyroxine be changed in hypothyroidism patients fasting during Ramadan?

Arzu OR KOCA, Murat DAĞDEVİREN, Mustafa ALTAY

Horse allergen sensitivity and respiratory symptoms among horse farm workers

Burcu BARAN KETENCİOĞLU, Nuri TUTAR, İnci GÜLMEZ, Fatma Sema OYMAK, İnsu YILMAZ

Özgür Korhan TUNÇEL, Gökhan SARISOY, Eda ÇETİN, Ebru KAYNAR TUNÇEL, Birşen BİLGİCİ, Arzu KARAUSTAOĞLU

Mohammad ZARE, Zahra TAGHARROBI, Khadıjeh SHARIFI, Zahra SOOKI, Javad ABOLHASANI

Immunomagnetic separation and Listeria monocytogenes detection with surfaceenhanced Raman scattering

Hilal TORUL, İsmail Hakkı BOYACI, Adem ZENGİN, Hande YEĞENOĞLU AKÇINAR, Belma ASLIM, Zekiye SULUDERE, Uğur TAMER, Burcu GÜVEN