Silibinin suppresses bladder cancer through down-regulation of actin cytoskeleton and PI3K/Akt signaling pathways.

Silibinin is the major active constituent of silymarin, an extract of milk thistle seeds. Silibinin has been shown to have significant anti-cancer effects in a variety of malignancies. However, the molecular mechanisms of silibinin action in bladder cancer have not been studied extensively. In the present study, we found that silibinin (10 μM) significantly suppressed proliferation, migration, invasion and induced apoptosis of T24 and UM-UC-3 human bladder cancer cells. Silibinin down-regulated the actin cytoskeleton and phosphatidylinositide 3-kinase (PI3K)/Akt signaling pathways in these cancer cell lines. These pathways were found to crosstalk through RAS cascades. We found that silibinin suppressed levels of trimethylated histone H3 lysine 4 and acetylated H3 at the KRAS promoter. Furthermore, silibinin targets long non-coding RNA: HOTAIR and ZFAS1, which are known to play roles as oncogenic factors in various cancers. This study shows that silibinin exerts anti-cancer effects through down-regulation of actin cytoskeleton and PI3K/Akt pathways and thus suppresses bladder cancer growth and progression

Tumor-suppressivemicroRNA-218inhibits cancer cell migration and invasion via targeting ofLASP1in prostate cancer

U uvodnom dijelu rada dana je definicija postupka toplinskog naštrcavanja. Detaljno su opisani značaj u pojedinim granama industrije, načini provedbe toplinskih naštrcavanja i primjena materijala. Opisane su najčešće vrste toplinskog naštrcavanja, te su nakon toga i međusobno uspoređene prema potrebnoj opremi za toplinsko naštrcavanje, zatim prema vrstama dodatnog materijala, te korištenom izvoru topline. U eksperimentalnom dijelu rada opisuje se plazma navarivanje kao nisko energetski postupak naštrcavanja, tj. brzina čestica je puno manja pa je sloj dodatnog materijala navaren. Analiziraju se načinjeni uzorci, ocjenjuje se prikladnost tehnologije i utvrđuju specifične greške prilikom postupka. U završnom dijelu rada dani su prijedlozi za poboljšanje tehnologije plazma navarivanja ovisno o vrsti praška i zahtjevima navarenog sloja

The MicroRNA Expression Signature of Bladder Cancer by Deep Sequencing: The Functional Significance of the <i>miR-195/497</i> Cluster

<div><p>Current genome-wide microRNA (miRNA) expression signature analysis using deep sequencing technologies can drive the discovery of novel cancer pathways regulated by oncogenic and/or tumor suppressive miRNAs. We determined the genome-wide miRNA expression signature in bladder cancer (BC) by deep sequencing technology. A total of ten small RNA libraries were sequenced (five BCs and five samples of histologically normal bladder epithelia (NBE)), and 13,190,619 to 18,559,060 clean small RNA reads were obtained. A total of 933 known miRNAs and 17 new miRNA candidates were detected in this analysis. Among the known miRNAs, a total of 60 miRNAs were significantly downregulated in BC compared with NBE. We also found that several miRNAs, such as <i>miR-1/133a</i>, <i>miR-206/133b</i>, <i>let-7c/miR-99a</i>, <i>miR-143/145</i> and <i>miR-195/497</i>, were located close together at five distinct loci and constituted clustered miRNAs. Among these clustered miRNAs, we focused on the <i>miR-195/497</i> cluster because this clustered miRNA had not been analyzed in BC. Transfection of mature <i>miR-195</i> or <i>miR-497</i> in two BC cell lines (BOY and T24) significantly inhibited cancer cell proliferation, migration and invasion, suggesting that the <i>miR-195/497</i> cluster functioned as tumor suppressors in BC. Regarding the genes targeted by the <i>miR-195/497</i> cluster, the TargetScan algorithm showed that 6,730 genes were putative <i>miR-195/497</i> targets, and 113 significantly enriched signaling pathways were identified in this analysis. The “Pathways in cancer” category was the most enriched, involving 104 candidate target genes. Gene expression data revealed that 27 of 104 candidate target genes were actually upregulated in BC clinical specimens. Luciferase reporter assays and Western blotting demonstrated that <i>BIRC5</i> and <i>WNT7A</i> were directly targeted by <i>miR-195/497</i>. In conclusion, aberrant expression of clustered miRNAs was identified by deep sequencing, and downregulation of <i>miR-195/497</i> contributed to BC progression and metastasis. Tumor suppressive miRNA-mediated cancer pathways provide new insights into the potential mechanisms of BC oncogenesis.</p></div

Patient characteristics for deep sequencing.

<p>Patient characteristics for deep sequencing.</p

Genistein inhibits prostate cancer cell growth by targeting miR-34a and oncogenic HOTAIR.

Genistein is a soy isoflavone that has antitumor activity both in vitro and in vivo. It has been shown that genistein inhibits many type of cancers including prostate cancer (PCa) by regulating several cell signaling pathways and microRNAs (miRNAs). Recent studies suggest that the long non-coding RNAs (lncRNAs) are also involved in many cellular processes. At present there are no reports about the relationship between gensitein, miRNAs and lncRNAs. In this study, we focused on miRNAs, lncRNA that are regulated by genistein and investigated their functional role in PCa.Microarray (SurePrint G3 Human GE 8×60K) was used for expression profiling of genistein treated and control PCa cells (PC3 and DU145). Functional assay (cell proliferation, migration, invasion, apoptosis and cell cycle assays) were performed with the PCa cell lines, PC3 and DU145. Both in vitro and in vivo (nude mouse) models were used for growth assays. Luciferase reporter assays were used for binding of miR-34a to HOTAIR.LncRNA profiling showed that HOTAIR was highly regulated by genistein and its expression was higher in castration-resistant PCa cell lines than in normal prostate cells. Knockdown (siRNA) of HOTAIR decreased PCa cell proliferation, migration and invasion and induced apoptosis and cell cycle arrest. miR-34a was also up-regulated by genistein and may directly target HOTAIR in both PC3 and DU145 PCa cells.Our results indicated that genistein inhibited PCa cell growth through down-regulation of oncogenic HOTAIR that is also targeted by tumor suppressor miR-34a. These findings enhance understanding of how genistein regulates lncRNA HOTAIR and miR-34a in PCa

Downregulated miRNAs in deep sequencing of BC.

<p>Downregulated miRNAs in deep sequencing of BC.</p