Previously, we designed a DNAzyme (b1DE) targeting the human b1 integrin subunit, which efficiently digested the mRNA of the b1 integrin subunit and downregulated b1 integrin expression in endothelial cells. This DNAzyme blocked the adhesion of endothelial cells and abolished their ability to form microcapillary tubes in Matrigel. In our present study, we demonstrate that b1DE effectively inhibited neovascularization in Matrigel plugs (BALB/c mice, n ¼ 20) and solid human carcinoma tumors developed in nude mice (BALB/cA nude (nu-/-)-B6.Cg-Foxn1 nu ) (n ¼ 30) using prostate carcinoma cells PC-3 (n ¼ 15) and colon adenocarcinoma cells CX1.1 (n ¼ 15). When injected intratumorally, it significantly reduced the tumor size and number of microvessels developed by both CX1.1 and PC-3 cells within the 3 weeks of experiment duration. Thus, DNAzymes targeting b1 integrin genes can inhibit multiple key tumorigenic processes in vitro and in vivo and may serve as useful anti-cancer agents.

The increased expression of TGFbeta1, MMP-9 observed in colorectal adenomas seems to be related to the grade of dysplasia. We assume that overexpression of TGFbeta1, MMP-9 represent an early event in colorectal carcinogenesis and may possibly have the prognostic value.

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Suppressor of cytokine signaling 1 (SOCS1) is the key regulator of cytokine-mediated innate and adaptive immunity. One of the molecular mechanisms of SOCS1 is connected with inhibition of TLR4-NFκB pathway. The relationships among these molecules in laryngeal carcinoma are not exactly known. In this preliminary study we focused on their special activity and role in regulation of development and progression of laryngeal carcinoma. To investigate NFκB (p65 subunit) nuclear and cytoplasmic expression in 45 tumor samples of advanced laryngeal carcinoma IHC staining was performed. To determine the mRNA expression levels of TLR4, IRAK1, TRAF6 and SOCS1 in isolated neoplasm cells and non-cancerous adjacent mucosa epithelial cells RT-PCR was used. The invasiveness of laryngeal carcinomas was evaluated according to tumor front grading, TFG, which included tumor-related features (cytoplasmic differentiation, nuclear polymorphism, number of mitoses) and adjacent stroma-related characteristics of the peripheral edge of tumor infiltration (mode of infiltration, depth of invasion and plasmalymphocytic infiltration). The relationships between pT, pN status, the histological G grade, certain clinicopathological characteristics as well as postoperative observation time and the mRNA expression of the molecules mentioned earlier were investigated. Significant differences of TLR4-NFκB pathway molecules and SOCS1 mRNA expression in laryngeal tumor cells and normal adjacent mucosa cells as well as significant interconnections of TLR4, SOCS1 and NFκB(p65) in isolated tumor cells were obtained. This preliminary study demonstrated that the expression of SOCS1 and TLR4-NFκB pathway molecules had a strong association with the aggressiveness of laryngeal carcinoma. Positive relationships of TRAF6 in tumor margin cells with the histological grade and the mode of tumor invasion as well as the TFG total score were highlighted. Significant positive correlations were found between the TLR4 in tumor central cells and the TFG total score. Negative relationships of SOCS1 in tumor central cells with the histological grade were also noted. Significant positive correlations were found between the cytoplasmic NFκB(p65) and the mode of invasion as well as TFG total score. Our findings confirmed the importance of SOCS1 and TLR4-NFκB pathway molecules as potential biomarkers for assessment of the aggressive tumor phenotype in laryngeal carcinoma.Key words: laryngeal carcinoma, SOCS1 (suppressor of cytokine signaling 1), TLR4 (toll-like receptor 4), IRAK1 (interleukin -1 receptor -associated kinase), TRAF6 (TNF receptor-associated factor 6), NFκB (nuclear factor kappa B, p65 subunit), TFG (tumor front grading)

Noradrenaline (NA) levels in pineal gland of chickens at various posthatching stages (P2, P4, P8, P15, P30 and P57) were determined by high-performance liquid chromatography with electrochemical detection. Pineal NA content markedly increased between P2 and P30. P30 and P57 chickens, kept from the day of hatching under a 12:12 hr light-dark (LD) illumination cycle, exhibited rhythmic changes in pineal NA, with levels in the dark period being markedly higher than in the light period. In younger birds pineal NA concentrations did not show pronounced daily variations. In 4-wk-old chickens (P28-30) kept under constant darkness (DD), the rhythmic pattern of pineal NA persisted for 1 day (with higher values during the subjective dark phase than during the subjective light phase), but this disappeared 24 hr after the introduction of DD. In contrast, NA content in pineal glands isolated from birds maintained for 2 days under continuous light was similar to that found during the light phase of the LD cycle, and did not exhibit significant rhythmicity. In P30 chickens, pretreated with alpha-methyl-p-tyrosine (AMPT, an inhibitor of tyrosine hydroxylase, the key regulatory enzyme in the biosynthesis of catecholamines), pineal NA content declined slowly and monophasically during the light phase. During the dark phase the AMPT-induced decay of NA was biphasic--namely an initial rapid decline over the first 15 min which was followed by a slow-rate decline--an observation indicating that NA turnover was higher in the dark. Acute exposure of the dark-adapted P30 and P57 chickens to light significantly decreased pineal NA content, but did not affect pineal NA concentrations in younger birds. Our results suggest that the NA rhythm in the chicken pineal gland and its sensitivity to light regulation progressively develop during the first month of life.