Genistein downregulates onco-miR-1260b and upregulates sFRP1 and Smad4 via demethylation and histone modification in prostate cancer cells

Background:Recently several microRNAs (miRNAs) have been found to be regulated by genistein in cancer cells. In this study, we focused on the gene regulatory effect of genistein on microRNA and its target genes in prostate cancer (PC).Methods:Initially, we investigated the effect of genistein on prostate cancer cells and identified that the expression of miRNA-1260b was decreased by genistein. We performed functional analyses and investigated the relationship between miRNA-1260b expression and prostate cancer patient outcomes. Two target genes (sFRP1 and Smad4) of miR-1260b were identified based on computer algorithm and 3′UTR luciferase assay was carried out to determine direct miRNA regulation of the genes.Results:Genistein promoted apoptosis while inhibiting prostate cancer cell proliferation, invasion and TCF reporter activity in PC cells. MiR-1260b was highly expressed in prostate cancer tissues and significantly downregulated by genistein in PC cells. After knocking down miR-1260b, cell proliferation, invasion, migration and TCF reporter activity were decreased in PC cells. Western analysis and 3′UTR luciferase assay showed that the two target genes (sFRP1 and Smad4) were directly regulated by miR-1260b. The expression of sFRP1 and Smad4 was significantly decreased in prostate cancer tissues. Genistein also increased expression of these two genes via DNA demethylation and histone modifications.Conclusions:Our data suggest that genistein exerts its anti-tumour effect via downregulation of miR-1260b that targeted sRRP1 and Smad4 genes in prostate cancer cells. The expression of sFRP1 and Smad4 was also modulated by genistein via DNA methylation or histone modifications in PC cell lines.

[1]  L. Wilkens,et al.  Cancer incidence trends among Asian American populations in the United States, 1990-2008. , 2013, Journal of the National Cancer Institute.

[2]  Yiwei Li,et al.  Inhibition of Nuclear Factor κB Activation in PC3 Cells by Genistein Is Mediated via Akt Signaling Pathway , 2002 .

[3]  N. Dubrawsky Cancer statistics , 1989, CA: a cancer journal for clinicians.

[4]  Ximing J. Yang,et al.  MEK4 function, genistein treatment, and invasion of human prostate cancer cells. , 2009, Journal of the National Cancer Institute.

[5]  L. Jian Soy, isoflavones, and prostate cancer. , 2009, Molecular nutrition & food research.

[6]  Stefano Piccolo,et al.  MicroRNA control of signal transduction , 2010, Nature Reviews Molecular Cell Biology.

[7]  Joachim Diebold,et al.  Functional epigenomics identifies genes frequently silenced in prostate cancer. , 2005, Cancer research.

[8]  J. Kench,et al.  Loss of BMP2, Smad8, and Smad4 expression in prostate cancer progression , 2004, The Prostate.

[9]  R. Lambrot,et al.  Histone methylation is a critical regulator of the abnormal expression of POU5F1 and RASSF1A in testis cancer cell lines. , 2011, International journal of andrology.

[10]  D. Cutler,et al.  Genistein cooperates with the histone deacetylase inhibitor vorinostat to induce cell death in prostate cancer cells , 2012, BMC Cancer.

[11]  M. Spitz,et al.  Limiting the location of putative human prostate cancer tumor suppressor genes on chromosome 18q , 1999, Oncogene.

[12]  A. Menke,et al.  Smad4-Independent TGF-β Signaling in Tumor Cell Migration , 2007, Cells Tissues Organs.

[13]  M. Wolter,et al.  Frequent promoter hypermethylation of Wnt pathway inhibitor genes in malignant astrocytic gliomas , 2010, International journal of cancer.

[14]  K. Katanoda,et al.  An updated report of the trends in cancer incidence and mortality in Japan. , 2013, Japanese journal of clinical oncology.

[15]  B. Bao,et al.  Perspectives on the Role of Isoflavones in Prostate Cancer , 2013, The AAPS Journal.

[16]  A. Perry,et al.  Discovery of DNA Hypermethylation Using a DHPLC Screening Strategy , 2007, Epigenetics.

[17]  Y. Bignon,et al.  Soy phytoestrogens modify DNA methylation of GSTP1, RASSF1A, EPH2 and BRCA1 promoter in prostate cancer cells. , 2010, In vivo.

[18]  I. Eltoum,et al.  Genistein in the diet reduces the incidence of poorly differentiated prostatic adenocarcinoma in transgenic mice (TRAMP). , 2001, Cancer research.

[19]  H. Akaza,et al.  Prostate Cancer Working Group report. , 2010, Japanese journal of clinical oncology.

[20]  A. Jemal,et al.  Global Cancer Statistics , 2011 .

[21]  L. Wilkens,et al.  Cancer incidence trends among native Hawaiians and other Pacific Islanders in the United States, 1990-2008. , 2013, Journal of the National Cancer Institute.

[22]  Michael Q. Zhang,et al.  Combinatorial patterns of histone acetylations and methylations in the human genome , 2008, Nature Genetics.

[23]  Long-Cheng Li,et al.  MethPrimer: designing primers for methylation PCRs , 2002, Bioinform..

[24]  I. Eltoum,et al.  Genistein alters growth but is not toxic to the rat prostate. , 2002, The Journal of nutrition.

[25]  E. Rimm,et al.  Diet After Diagnosis and the Risk of Prostate Cancer Progression, Recurrence, and Death (United States) , 2006, Cancer Causes & Control.

[26]  F. Bosman,et al.  Alterations of the Wnt signaling pathway during the neoplastic progression of Barrett's esophagus , 2006, Oncogene.

[27]  A. Jemal,et al.  Cancer statistics, 2012 , 2012, CA: a cancer journal for clinicians.

[28]  T. Swigut,et al.  H3K27 Demethylases, at Long Last , 2007, Cell.

[29]  R. Dahiya,et al.  Genistein Up-Regulates Tumor Suppressor MicroRNA-574-3p in Prostate Cancer , 2013, PloS one.

[30]  G. Morgia,et al.  Polyphenols: Key Issues Involved in Chemoprevention of Prostate Cancer , 2012, Oxidative medicine and cellular longevity.

[31]  A. Menke,et al.  Smad4-independent TGF-beta signaling in tumor cell migration. , 2007, Cells, tissues, organs.

[32]  Y. Bignon,et al.  miRNAs differentially expressed in prostate cancer cell lines after soy treatment. , 2011, In vivo.

[33]  Yiwei Li,et al.  Inhibition of nuclear factor kappaB activation in PC3 cells by genistein is mediated via Akt signaling pathway. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[34]  A. Svindland,et al.  Efficacy and Safety of Short-Term Genistein Intervention in Patients with Localized Prostate Cancer Prior to Radical Prostatectomy: A Randomized, Placebo-Controlled, Double-Blind Phase 2 Clinical Trial , 2011, Nutrition and cancer.

[35]  E. Fearon,et al.  Cancer progression , 1999, Current Biology.

[36]  M. Bosland,et al.  Chemoprevention of prostate cancer: Natural compounds, antiandrogens, and antioxidants – In vivo evidence , 2011, Journal of carcinogenesis.

[37]  R. Dahiya,et al.  Genistein downregulates onco-miR-1260b and inhibits Wnt-signalling in renal cancer cells , 2013, British Journal of Cancer.

[38]  Scott E. Kern,et al.  DPC4, A Candidate Tumor Suppressor Gene at Human Chromosome 18q21.1 , 1996, Science.

[39]  A. Jemal,et al.  Global cancer statistics , 2011, CA: a cancer journal for clinicians.

[40]  T. H. van der Kwast,et al.  Prevalence, treatment modalities and prognosis of familial prostate cancer in a screened population. , 2006, The Journal of urology.

[41]  K. Kinzler,et al.  Frequency of Smad gene mutations in human cancers. , 1997, Cancer research.

[42]  Hui Wang,et al.  Genistein, a dietary isoflavone, down-regulates the MDM2 oncogene at both transcriptional and posttranslational levels. , 2005, Cancer research.

[43]  Yoshiaki Kawano,et al.  Secreted antagonists of the Wnt signalling pathway , 2003, Journal of Cell Science.

[44]  T. Ørntoft,et al.  Non‐CpG island promoter hypomethylation and miR‐149 regulate the expression of SRPX2 in colorectal cancer , 2013, International journal of cancer.

[45]  John C. Boik,et al.  A comparative survey of leguminous plants as sources of the isoflavones, genistein and daidzein: implications for human nutrition and health. , 1997, Journal of alternative and complementary medicine.

[46]  Gerald C. Chu,et al.  SMAD4-dependent barrier constrains prostate cancer growth and metastatic progression , 2011, Nature.

[47]  S. Shukla,et al.  Dietary Agents in the Chemoprevention of Prostate Cancer , 2005, Nutrition and cancer.

[48]  Jun Wang,et al.  Genistein alters growth factor signaling in transgenic prostate model (TRAMP) , 2004, Molecular and Cellular Endocrinology.

[49]  Wei Dong Chen,et al.  Epigenetic inactivation of SFRP genes allows constitutive WNT signaling in colorectal cancer , 2004, Nature Genetics.