The Effect of Snail1 Gene Silencing by siRNA in Metastatic Breast Cancer Cell Lines
Abstract
Background: Breast cancer is the most common diagnosed cancer among women in the world. Snail1 plays a role in the development of the invasive phenotypes of cancer, neural cell differentiation, cell division and apoptosis in tumor cells. Traces of snail1 in metastasis of breast cancer to bone are observed. The aim of this study was to investigate the effect of specific snail1 siRNAs on the proliferation, migration, induction of apoptosis and cell cycle arrest of MDA-MB-468 cells.
Methods: In 2015, this experimental study was performed on the MDA-MB-468 cell lines in Immunology Research Center, Tabriz University of Medical Sciences. After the design and construction of siRNA, transfection was performed with transfection reagent. The expression levels of mRNA and protein were measured by qRT-PCR and western blot analysis, respectively. The survival of cells was determined by using MTT assay cells, apoptosis using Tunel assay, Cell migration using scratch assay, Cell cycle analysis by Propidium Iodide (PI) DNA staining method using flow cytometry on the MDA-MB-468.
Results: Transfection with siRNA significantly suppressed the expression of snail1 gene in dose-dependent manner after 48 h (P<0.0001). Surprisingly, treatment with snail1 siRNA arrested cell cycle in S phases (P<0.0001). Moreover, siRNA transfection had effects on breast adenocarcinoma cells and inhibited the migration (P<0.0001), proliferation (P<0.0001) and induced apoptosis (P<0.0016).
Conclusion: The snail1 can be considered as a potent adjuvant in breast cancer therapy.
Sahranavard S, Naghibi F, Mosaddegh M, Esmaeili S, Sarkhail P, Taghvaei M, Ghafari S (2009). Cytotoxic activities of selected medicinal plants from Iran and phytochemical evaluation of the most potent extract. Res Pharm Sci,4(2):133-7.
Valdivieso M, Kujawa AM, Jones T, Baker LH (2012). Cancer survivors in the United States: a review of the literature and a call to action. Int J Med Sci,9(2):163-73.
Winer E, Gralow J, Diller L et al (2009). Clinical cancer advances 2008: major research advances in cancer treatment, prevention, and screening—a report from the American Society of Clinical Oncology. J Clin Oncol,27(5):812-26.
DeSantis C, Ma J, Bryan L, Jemal A (2014). Breast cancer statistics, 2013. CA Cancer J Clin,64(1):52-62.
Aamir Ahmad. (2013). Breast Cancer Metastasis and Drug Resistance Progress and Prospects. New York.
Izquierdo A, Gispert R, Saladie F, Espinàs JA (2008). Análisis de la incidencia, la supervivencia y la mortalidad según las principales localizaciones tumorales, 1985-2019: cáncer de mama. Med Clin,131:50-2.
Yun J, Frankenberger CA, Kuo WL et al (2011). Signalling pathway for RKIP and Let‐7 regulates and predicts metastatic breast cancer. EMBO J,30(21):4500-14.
Baritaki S, Chapman A, Yeung K, Spandidos D, Palladino M, Bonavida B (2009). Inhibition of epithelial to mesenchymal transition in metastatic prostate cancer cells by the novel proteasome inhibitor, NPI-0052: pivotal roles of Snail repression and RKIP induction. Oncogene,28(40):3573-85.
Baritaki S, Huerta-Yepez S, Sakai T, Spandidos DA, Bonavida B (2007). Chemotherapeutic drugs sensitize cancer cells to TRAIL-mediated apoptosis: up-regulation of DR5 and inhibition of Yin Yang 1. Mol Cancer Ther,6(4):1387-99.
Jordà M, Vinyals A, Marazuela A et al (2007). Id-1 is induced in MDCK epithelial cells by activated Erk/MAPK pathway in response to expression of the Snail and E47 transcription factors. Exp Cell Res,313(11):2389-403.
De Craene B, Gilbert B, Stove C, Bruyneel E, Van Roy F, Berx G (2005). The transcription factor snail induces tumor cell invasion through modulation of the epithelial cell differentiation program. Cancer Res,65(14):6237-44.
Peinado H, Olmeda D, Cano A (2007). Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nature Reviews Cancer,7(6):415-28.
Thiery JP, Acloque H, Huang RY, Nieto MA (2009). Epithelial-mesenchymal transitions in development and disease. Cell,139(5):871-90.
Thuault S, Tan E-J, Peinado H, Cano A, Heldin C-H, Moustakas A (2008). HMGA2 and Smads co-regulate SNAIL1 expression during induction of epithelial-to-mesenchymal transition. J Biol Chem,283(48):33437-46.
Yuan H, Kajiyama H, Ito S et al (2013). ALX1 induces snail expression to promote epithelial-to-mesenchymal transition and invasion of ovarian cancer cells. Cancer Res,73(5):1581-90.
Sun M, Guo X, Qian X et al (2012). Activation of the ATM-Snail pathway promotes breast cancer metastasis. J Mol Cell Biol,4(5):304-15.
Zhang A, Chen G, Meng L et al (2008). Antisense-Snail transfer inhibits tumor metastasis by inducing E-cadherin expression. Anticancer Res,28(2A):621-8.
Olmeda D, Jorda M, Peinado H, Fabra A, Cano A (2007). Snail silencing effectively suppresses tumour growth and invasiveness. Oncogene,26(13):1862-74.
Sugimachi K, Tanaka S, Kameyama Tet al (2003). Transcriptional repressor snail and progression of human hepatocellular carcinoma. Clin Cancer Res,9(7):2657-64.
Kanellopoulou C, Monticelli S (2008). A role for microRNAs in the development of the immune system and in the pathogenesis of cancer. Semin Cancer Biol,18(2):79-88.
Giovannetti E, Erozenci A, Smit J, Danesi R, Peters GJ (2012). Molecular mechanisms underlying the role of microRNAs (miRNAs) in anticancer drug resistance and implications for clinical practice. Crit Rev Oncol Hematol,81(2):103-22.
Li M, Li J, Ding X, He M, Cheng SY (2010). microRNA and cancer. AAPS J,12(3):309-17.
Hickey M, Peate M, Saunders CM, Friedlander M (2009). Breast cancer in young women and its impact on reproductive function. Hum Reprod Update,15(3):323-39.
Tavakoli J, Miar S, Majid Zadehzare M, Akbari H (2012). Evaluation of effectiveness of herbal medication in cancer care: a review study. Iran J Cancer Prev,5(3):144-56.
Karami H, Baradaran B, Esfahani A, Sakhinia M, Sakhinia E (2014). Therapeutic Effects of Myeloid Cell Leukemia-1 siRNA on Human Acute Myeloid Leukemia Cells. Adv Pharm Bull,4(3):243-8.
Zaffaroni N, Daidone MG (2002). Survivin expression and resistance to anticancer treatments: perspectives for new therapeutic interventions. Drug Resist Updat,5(2):65-72.
Gritsko T, Williams A, Turkson J et al (2006). Persistent activation of stat3 signaling induces survivin gene expression and confers resistance to apoptosis in human breast cancer cells. Clin Cancer Res,12(1):11-9.
Blanco MJ, Moreno-Bueno G, Sarrio D, Locascio A, Cano A, Palacios J, Nieto MA (2002). Correlation of Snail expression with histological grade and lymph node status in breast carcinomas. Oncogene,21(20):3241-6.
Imai T, Horiuchi A, Wang C, Oka K, Ohira S, Nikaido T, Ikuo Konishi (2003). Hypoxia attenuates the expression of E-cadherin via up-regulation of SNAIL in ovarian carcinoma cells. Am J Pathol,163(4):1437-47.
Poser I, Domı́nguez D, de Herreros AG, Varnai A, Buettner R, Bosserhoff AK (2001). Loss of E-cadherin expression in melanoma cells involves up-regulation of the transcriptional repressor Snail. J Biol Chem,276(27):24661-6.
Takkunen M, Grenman R, Hukkanen M, Korhonen M, De Herreros AG, Virtanen I (2006). Snail-dependent and-independent epithelial-mesenchymal transition in oral squamous carcinoma cells. J Histochem Cytochem,54(11):1263-75.
Becker K-F, Rosivatz E, Blechschmidt K, Kremmer E, Sarbia M, Höfler H (2007). Analysis of the E-cadherin repressor Snail in primary human cancers. Cells Tissues Organs,185(1-3):204-12.
Espineda CE, Chang JH, Twiss J, Rajasekaran SA, Rajasekaran AK (2004). Repression of Na, K-ATPase β1-subunit by the transcription factor snail in carcinoma. Mol Biol Cell,15(3):1364-73.
Barrallo-Gimeno A, Nieto MA (2005). The Snail genes as inducers of cell movement and survival: implications in development and cancer. Development,132(14):3151-61.
Peinado H, Portillo F, Cano A (2004). Transcriptional regulation of cadherins during development and carcinogenesis. Int J Dev Biol, 48(5-6):365-75.
Thiery JP, Sleeman JP (2006). Complex networks orchestrate epithelial–mesenchymal transitions. Nat Rev Mol Cell Biol,7(2):131-42.
Wan Z, Pan H, Liu S, Zhu J, Qi W, Fu K, Zhao T, Liang J (2015). Downregulation of SNAIL sensitizes hepatocellular carcinoma cells to TRAIL-induced apoptosis by regulating the NF-κB pathway. Oncol Rep,33(3):1560-6.
Osorio LA, Farfán NM, Castellón EA, Contreras HR (2016). SNAIL transcription factor increases the motility and invasive capacity of prostate cancer cells. Mol Med Rep,13(1):778-86.
Neal CL, Mckeithen D, Odero-Marah VA (2011). Snail negatively regulates cell adhesion to extracellular matrix and integrin expression via the MAPK pathway in prostate cancer cells. Cell Adh Migr,5(3):249-57.
Jin H, Yu Y, Zhang T et al (2010). Snail is critical for tumor growth and metastasis of ovarian carcinoma. Int J Cancer,126(9):2102-11.
de Souza Palma C, Grassi ML, Thomé CH (2016). Proteomic analysis of epithelial to mesenchymal transition reveals crosstalk between SNAIL and HDAC1 in breast cancer cells. Mol Cell Proteomics, 15(3):906-17.
Lundgren K, Nordenskjöld B, Landberg G (2009). Hypoxia, Snail and incomplete epithelial–mesenchymal transition in breast cancer. Br J Cancer,101(10):1769-81.
Vega S, Morales AV, Ocaña OH, Valdés F, Fabregat I, Nieto MA (2004). Snail blocks the cell cycle and confers resistance to cell death. Genes Dev,18(10):1131-43.
Andorfer CA, Necela BM, Thompson EA, Perez EA (2011). MicroRNA signatures: clinical biomarkers for the diagnosis and treatment of breast cancer. Trends Mol Med,17(6):313-9.
Heneghan H, Miller N, Lowery A, Sweeney K, Kerin M (2009). MicroRNAs as novel biomarkers for breast cancer. J Oncol, 950201.
Fu SW, Chen L, Man Y (2011). miRNA biomarkers in breast cancer detection and management. J Cancer,2:116-22.
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| Issue | Vol 46 No 5 (2017) | |
| Section | Original Article(s) | |
| Keywords | ||
| Snail1 siRNA RNA interference Cell line Breast cancer | ||
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