There is a growing body of evidence suggesting antitumor activity of statins. In metastasis and invasion of cancer the Epithelial–Mesenchymal Transition (EMT) of cancerous cells is an important process. Our goal was to understand the effect of Rosuvastatin on the EMT process in human prostate cancer cell line PC-3 cells in adherent 2 dimensional (2D) and spheroid 3 dimensional (3D) culture. PC-3 cells were cultured in adherence and/or spheroid culture system. The cells were treated with different concentrations of Rosuvastatin. After 96 h, the cell proliferation, viability, type and number of spheroids, the expression of E-Cadherin, Vimentin and Zeb-1 were analyzed. The results show that Rosuvastatin inhibit cell proliferation without significant cytotoxicity. The spheroid formation and spheroid sizes were inhibited by Rousavastatin in a dose dependent manner. In 2D culture, expression of the E-Cadherin was increased up to 2.0 fold in a dose dependent linear manner (R2 = 0.89). Vimentin and Zeb-1 expressions were decreased up to 40 and 20% of untreated control cells expression level respectively, (R2 = 0.99 and 0.92). In 3D system, the expression of E-Cadherin did not show a significant change, but Vimentin and Zeb-1 expressions were decreased up to 70 and 40% of untreated control cells expression level respectively in a dose dependent linear manner in comparison to 2D system (R2 = 0.36 and 0.90). Our finding indicates that Rousavastatin inhibit cell proliferation and spheroid formation of PC-3 cells. This inhibition accompanies by inhibition of EMT markers. Therefor, this cholesterol lowering agent could probably have potential in the prevention and suppression of cancer in androgen dependent prostate cancer.
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Marín de Mas I, Aguilar E, Jayaraman A, Polat IH, Martín-Bernabé A et al (2014) Cancer cell metabolism as new targets for novel designed therapies. Future Med Chem 6(16):1791–1810
Alderton GK (2014) Metastasis: metabolic reprogramming in disseminated cells. Nat Rev Cancer 14(11):703. https://doi.org/10.1038/nrc3842
Hryniewicz-Jankowska A, Augoff K, Sikorski AF (2019) Highlight article: the role of cholesterol and cholesterol-driven membrane raft domains in prostate cancer. Exp Biol Med (Maywood) 244(13):1053–1061. https://doi.org/10.1177/1535370219870771
Shitara Y, Sugiyama Y (2006) Pharmacokinetic and pharmacodynamic alterations of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors: drug–drug interactions and interindividual differences in transporter and metabolic enzyme functions. Pharmacol Ther 112(1):71–105. https://doi.org/10.1016/j.pharmthera.2006.03.003
Yeganeh B, Wiechec E, Ande SR, Sharma P, Moghadam AR et al (2014) Targeting the mevalonate cascade as a new therapeutic approach in heart disease, cancer and pulmonary disease. Pharmacol Ther 143(1):87–110. https://doi.org/10.1016/j.pharmthera
Janicko M, Drazilova S, Pella D, Fedacko J, Jarcuska P (2016) Pleiotropic effects of statins in the diseases of the liver. World J Gastroenterol 22(27):6201–6213. https://doi.org/10.3748/wjg.v22.i27.6201
Pavan LMC, Rêgo DF, Elias ST, De Luca Canto G, Silva Guerra EN (2015) In vitro anti-tumor effects of statins on head and neck squamous cell carcinoma: a systematic review. PLoS ONE 10(6):e0130476. https://doi.org/10.1371/journal.pone
Lytras T, Nikolopoulos G, Bonovas S (2014) Statins and the risk of colorectal cancer: an updated systematic review and meta-analysis of 40 studies. World J Gastroenterol 20(7):1858–1870. https://doi.org/10.3748/wjg.v20.i7
Hamada T, Khalaf N, Yuan C, Morales-Oyarvide V, Babic A, Nowak JA et al (2018) Pre-diagnosis use of statins associates with increased survival times of patients with pancreatic cancer. Clin Gastroenterol Hepatol 16(8):1300-1306.e3. https://doi.org/10.1016/j.cgh
Omori M, Okuma Y, Hakozaki T, Hosomi Y (2019) Statins improve survival in patients previously treated with nivolumab for advanced non-small cell lung cancer: an observational study. Mol Clin Oncol 10(1):137–143. https://doi.org/10.3892/mco
Skaletz-Rorowski A, Walsh K (2003) Statin therapy and angiogenesis. Curr Opin Lipidol 14(6):599–603
Dulak J, Jozkowicz A (2005) Anti-angiogenic and anti-inflammatory effects of statins: relevance to anti-cancer therapy. Curr Cancer Drug Targets 5(8):579–594. https://doi.org/10.2174/156800905774932824
Clendening JW, Penn LZ (2012) Targeting tumor cell metabolism with statins. Oncogene 31(48):4967–4978. https://doi.org/10.1038/onc.2012.6
Hindler K et al (2006) The role of statins in cancer therapy. Oncologist 11(3):306–315. https://doi.org/10.1634/theoncologist.11-3-306
Micalizzi DS, Farabaugh SM, Ford HL (2010) Epithelial–mesenchymal transition in cancer: parallels between normal development and tumor progression. J Mammary Gland Biol Neoplasia 15(2):117–134. https://doi.org/10.1007/s10911-010-9178-9
Gravdal K et al (2007) A switch from E-cadherin to N-cadherin expression indicates epithelial to mesenchymal transition and is of strong and independent importance for the progress of prostate cancer. Clin Cancer Res 13(23):7003–7011. https://doi.org/10.1158/1078-0432.CCR-07-1263
Grant CM, Kyprianou N (2013) Epithelial mesenchymal transition (EMT) in prostate growth and tumor progression. Transl Androl Urol 2(3):202–211. https://doi.org/10.3978/j.issn.2223-4683.2013.09.04
Di Zazzo E, Galasso G, Giovannelli P, Di Donato M, Bilancio A et al (2019) Estrogen receptors in epithelial–mesenchymal transition of prostate cancer. Cancers (Basel) 11(10):1418. https://doi.org/10.3390/cancers11101418
Rossi V, Di Zazzo E, Galasso G, De Rosa C, Abbondanza C et al (2019) Estrogens modulate somatostatin receptors expression and synergize with the somatostatin analog pasireotide in prostate cells. Front Pharmacol 10:1–8. https://doi.org/10.3389/fphar.2019.00028
Achilli T-M, Meyer J, Morgan JR (2012) Advances in the formation, use and understanding of multi-cellular spheroids. Expert Opin Biol Ther 12(10):1347–1360. https://doi.org/10.1517/14712598.2012.707181
Elliott NT, Yuan F (2011) A review of three-dimensional in vitro tissue models for drug discovery and transport studies. J Pharm Sci 100(1):59–74. https://doi.org/10.1002/JPS.22257
Williams C, Xie AW, Yamato M, Okana T, Wong JY (2011) ‘Stacking of aligned cell sheets for layer-by-layer control of complex tissue structure. Biomaterial 32(24):5625–5632
Duguay D, Foty RA, Steinberg MS (2003) Cadherin-mediated cell adhesion and tissue segregation: qualitative and quantitative determinants. Dev Biol 253(2):309–323. https://doi.org/10.1016/S0012-1606(02)00016-7
Ghulam J, Stuerken C, Wicklein D, Pries R, Wollenberg B, Schumacher U (2019) Immunohistochemical analysis of transcription factors and markers of epithelial–mesenchymal transition (EMT) in human tumors. Anticancer Res 39(10):5437–5448. https://doi.org/10.21873/anticanres.13737
Iwatsuki M, Mimori K, Yokobori T, Ishi H, Beppu T, Nakamori S et al (2010) Epithelial–mesenchymal transition in cancer development and its clinical significance. Cancer Sci 101(2):293–299
Cheaito KA, Bahmad HF, Hadadeh O, Saleh E, Dagher C, Hammoud MS et al (2019) EMT markers in locally-advanced prostate cancer: predicting recurrence? Front Oncol 11(9):131. https://doi.org/10.3389/fonc.2019.00131
Mitra A, Satelli A, Xia X, Cutrera J, Mishra L, Li S (2015) Cell-surface Vimentin: a mislocalized protein for isolating csVimentin(+) CD133(-) novel stem-like hepatocellular carcinoma cells expressing EMT markers. Int J Cancer 137(2):491–496. https://doi.org/10.1002/ijc.29382
Saar M, Zhao H, Nolley R, Young SR, Coleman I et al (2014) Spheroid culture of LuCaP 147 as an authentic preclinical model of prostate cancer subtype with SPOP mutation and hypermutator phenotype. Cancer Lett 351(2):272–280. https://doi.org/10.1016/j.canlet.2014.06.014
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 22DDCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Moreno-Bueno G, Peinado H, Molina P, Olmeda D, Cubillo E, Santos V, Palacios J et al (2009) The morphological and molecular features of the epithelial-to-mesenchymal transition. Nat Protoc 4:1591–1613
Chen T, You Y, Jiang H, Wang ZZ (2017) Epithelial–mesenchymal transition (EMT): a biological process in the development, stem cell differentiation and tumorigenesis. J Cell Physiol 232(12):3261–3272
Qureshi-Baig K, Ullmann P, Rodriguez F, Frasquilho S, Nazarov PV, Haan S, Letellier E (2016) What do we learn from spheroid culture systems? Insights from tumorspheres derived from primary colon cancer tissue. PLoS ONE. https://doi.org/10.1371/journal.pone.0150179
Riemann A, Rauschner M, Gießelmann M, Reime S, Haupt V, Thews O (2019) Extracellular acidosis modulates the expression of epithelial–mesenchymal transition (EMT) markers and adhesion of epithelial and tumor cells. Neoplasia 21(5):450–458. https://doi.org/10.1016/j.neo.2019.03.004
Peinado H, Olmeda D, Cano A (2007) Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer 7:415–428
Drivalos A, Chrisofos M, Efstathiou E, Kapranou A, Kollaitis G et al (2016) Expression of _5-integrin, _7-integrin, E-cadherin, and N-cadherin in localized prostate cancer. Urol Oncol 34:e11–e18
Hotz B, Arndt M, Dullat S, Bahrgava S, Buhr HJ, Hotz HG (2007) Epithelial to mesenchymal transition: expression of the regulators snail, slug, and twist in pancreatic cancer. Clin Cancer Res 13:4769–4776
Celesti G, Di Caro G, Bianchi P, Grizzi F, Basso G et al (2013) Presence of Twist1-positive neoplastic cells in the stroma of chromosome-unstable colorectal tumors. Gastroenterology 145:647–657
Van Roy F, Berx G (2008) The cell-cell adhesion molecule E-cadherin. Cell Mol Life Sci 65:3756–3788
Jang HJ, Hong EM, Park SW, Byun HW, Koh DH et al (2016) (2016) Statin induces apoptosis of human colon cancer cells and downregulation of insulin-like growth factor 1 receptor via proapoptotic ERK activation. Oncol Lett 12(1):250–256. https://doi.org/10.3892/ol.2016.4569
Lee SK, Kim YS (2013) Phosphorylation of eIF2α attenuates statin-induced apoptosis by inhibiting the stabilization and translocation of p53 to the mitochondria. Int J Oncol 42(3):810–816. https://doi.org/10.3892/ijo.2013.1792
Saha B, Arase A, Imam SS, Tsao-Wei D, Naritoku WY et al (2008) Overexpression of E-cadherin and beta-catenin proteins in metastatic prostate cancer cells in bone. Prostate 68:78–84
Pontes J Jr, Srougi M, Borra PM, DallOglio MF, Ribeiro-Filho LA, Leite KR (2010) E-cadherin and beta-catenin loss of expression related to bone metastasis in prostate cancer. Appl Immunohistochem Mol Morphol 18:179–184
Stadler M, Scherzer M, Walter S, Holzner S, Pudelko K et al (2018) Exclusion from spheroid formation identifies loss of essential cell-cell adhesion molecules in colon cancer cells. Sci Rep 8:1151. https://doi.org/10.1038/s41598-018-19384-0
Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A et al (2008) The epithelial–mesenchymal transition generates cells with properties of stem cells. Cell 133:704–715
Fontana F, Raimondi M, Marzagalli M, Sommariva M, Limonta P, Gagliano N (2019) Epithelial-to-mesenchymal transition markers and CD44 isoforms are differently expressed in 2D and 3D cell cultures of prostate cancer cells. Cells 8(2):143–149. https://doi.org/10.3390/cells8020143
Gagliano N, Celesti G, Tacchini L, Pluchino S, Sforza C et al (2016) Epithelial-to-mesenchymal transition in pancreatic ductal adenocarcinoma: characterization in a 3D-cell culture model. World J Gastroenterol 22(18):4466–4483. https://doi.org/10.3748/wjg.v22.i18.4466
This work was supported by research funds (No. 619) from National Institute of Genetic Engineering and Biotechnology, Tehran-Iran.
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The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.
The study protocol was approved by the Research Ethics Committee at National Institute of Genetic Engineering and Biotechnology (Tehran-Iran).
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Deezagi, A., Safari, N. Rosuvastatin inhibit spheroid formation and epithelial–mesenchymal transition (EMT) in prostate cancer PC-3 cell line. Mol Biol Rep 47, 8727–8737 (2020). https://doi.org/10.1007/s11033-020-05918-1
- Prostate cancer
- Epithelial–mesenchymal transition
- Spheroid culture