Genistein downregulates onco-miR-1260b and inhibits Wnt-signalling in renal cancer cells

• Published in: British journal of cancer Vol. 108; no. 10; pp. 2070 - 2078
• Main Authors: HIRATA, H, UENO, K, NAKAJIMA, K, TABATABAI, Z. L, HINODA, Y, ISHII, N, DAHIYA, R
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 28.05.2013
• Subjects: Adult; Aged; Anticarcinogenic Agents - pharmacology; Apoptosis; Biological and medical sciences; Carcinoma, Renal Cell - genetics; Carcinoma, Renal Cell - metabolism; Carcinoma, Renal Cell - pathology; Cell culture; Cell cycle; Cell growth; Cell Proliferation; Down-Regulation - drug effects; Down-Regulation - genetics; Drug Evaluation, Preclinical; Female; Gene expression; Gene Expression Regulation, Neoplastic - drug effects; Genistein - pharmacology; Humans; Kidney cancer; Kidney Neoplasms - genetics; Kidney Neoplasms - metabolism; Kidney Neoplasms - pathology; Kidneys; Male; Medical sciences; Metastasis; MicroRNAs; MicroRNAs - genetics; Middle Aged; Molecular Diagnostics; Multiple tumors. Solid tumors. Tumors in childhood (general aspects); Nephrology. Urinary tract diseases; Oncogenes - drug effects; Oncogenes - genetics; Proteins; Tumor Cells, Cultured; Tumors; Tumors of the urinary system; Urology; Wnt Signaling Pathway - drug effects; San Francisco California; California; United States > US; Japan; miR-1260b; Smad protein; Carcinoma; Genistein; Tumorous infiltration; Metastasis; FACS; invasion assay; real-time RT-PCR; Isoflavone derivatives; Flavonoid; Assay; Signal transduction; Polyphenol; Cancerology; sFRP1; 3' UTR luciferase assay; Immunoblotting assay; Grawitz tumor; TOPflash; Kidney disease; MTS assay; Urinary system disease; Enzyme; Tyrosine kinase inhibitor; Luciferase; Malignant tumor; western blot; Invasion; Dkk2; Cytometry; SFRP1gene; RCC; Smad4; Cell death; Kidney cancer; Phenols; Oxidoreductases; Cancer; Apoptosis; Real time polymerase chain reaction
• ISSN: 0007-0920; 1532-1827
• DOI: 10.1038/bjc.2013.173

Wnt-signalling has an important role in renal cancer and it is modulated by genistein in other cancers. Recently, microRNAs (miRNAs) have emerged as new regulators of gene expression. Thus, we focused on miRNAs to examine the regulatory mechanism of genistein on the Wnt-signalling pathway in renal cell carcinoma (RCC). Initially, we investigated the effect of genistein on Wnt-signalling (TOPflash reporter assay (TCF reporter assays)) in renal cancer cells, and using microarray identified candidate miRNAs whose expression was decreased by genistein. We performed functional analyses and investigated the relationship between miRNA expression and renal cancer patient outcomes. We also did 3'UTR luciferase assays to look at direct miRNA regulation of Wnt-signalling-related genes. Genistein promoted apoptosis while inhibiting RCC cell proliferation and invasion. Genistein also decreased TCF reporter activity in RCC cells. We found that miR-1260b was highly expressed and significantly downregulated by genistein in RCC cells. The expression of miR-1260b was significantly higher in renal cancer tissues compared with normal, and significantly related to overall shorter survival. In addition, miR-1260b promoted renal cancer cell proliferation and invasion in RCC cells. The 3'UTR luciferase activity of target genes (sFRP1, Dkk2, Smad4) was significantly decreased and their protein expression significantly upregulated in miR-1260b inhibitor-transfected renal cancer cells. Our data suggest that genistein inhibited Wnt-signalling by regulating miR-1260b expression in renal cancer cells.