Expression and regulatory effects on cancer cell behavior of NELL1 and NELL2 in human renal cell carcinoma

We thank Professors Michael Rehli, Yoshiaki Ito, and Kristian Helin for gifting plasmids, Dr. Alasdair MacKenzie (University of Aberdeen) for helpful discussion, and Mr. Takashi Mizukami, Ms. Ryoko Tokuda, and Ms. Sanae Funaoka (Kanazawa University) for technical assistance.Peer reviewedPublisher PD

Structural and kinetic modification of aqueous hydroxypropylmethylcellulose(HPMC) induced by electron beam irration

Electron beam was irradiated on 10% and 20% hydroxypropylmethylcellulose (HPMC) aqueous solutions with different doses to make gel films. As increasing dose, the gel fraction of the film increased sharply above a critical dose and then decreased gradually after passing a maximum. The scission/cross-linking ratio and the critical dose were determined using the Charlesby-Rosiak equation as 0.52 and 9 kGy for 10% gel and 0.43 and 14 kGy for 20% gel, respectively. The gel fraction for 20% HPMC film was lower at low dose and higher at high dose than that for 10% film. The behavior of the swelling ratio of the gel film was just opposite to that of the gel fraction. The cross-linking density of the gel estimated from the Flory theory linearly increased with irradiation dose at low dose, passed a maximum around 100 and 160 kGy for 10% and 20% films, respectively, and decreased at high dose. These results suggest the competition of scission and cross-linking induced by indirect effect of irradiation. Dielectric relaxation measurement by time domain reflectometry and RF impedance/material analyzer revealed two characteristic relaxations of chain motions around 100MHz and of orientation of free water around 20GHz. From the dose dependence of the relaxation parameters determined by fitting to a combined equation of Cole-Cole type and KWW type, a coupling of motions of HPMC molecules and water molecules was strongly suggested. The critical dose for gelation was coincident with the dose for the maximum of t h and the minimum of Deh together with the minimum of t m and the maximum of Dem, where t h and Deh denote the relaxation time and the relaxation strength for free water molecular motion and t m and Dem the corresponding ones for HPMC molecular motion. The characteristic behavior was discussed in terms of the increase of affinity between HPMC and water and the constrained molecular motion in the gel network

Expression and regulatory effects on cancer cell behavior of NELL1 and NELL2 in human renal cell carcinoma

Neural epidermal growth factor-like like (NELL) 1 and 2 constitute a family of multimeric and multimodular extracellular glycoproteins. Although the osteogenic effects of NELL1 and functions of NELL2 in neural development have been reported, their expression and functions in cancer are largely unknown. In this study, we examined expression of NELL1 and NELL2 in renal cell carcinoma (RCC) using clinical specimens and cell lines. We show that, whereas NELL1 and NELL2 proteins are strongly expressed in renal tubules in non-cancerous areas of RCC specimens, their expression is significantly downregulated in cancerous areas. Silencing of NELL1 and NELL2 mRNA expression was also detected in RCC cell lines. Analysis of NELL1/2 promoter methylation status indicated that the CpG islands in the NELL1 and NELL2 genes are hypermethylated in RCC cell lines. NELL1 and NELL2 bind to RCC cells, suggesting that these cells express a receptor for NELL1 and NELL2 that can transduce signals. Furthermore, we found that both NELL1 and NELL2 inhibit RCC cell migration, and NELL1 further inhibits RCC cell adhesion. These results suggest that silencing of NELL gene expression by promoter hypermethylation plays roles in RCC progression by affecting cancer cell behavior. We found that the down-regulation of NELL1 and NELL2 in renal cell carcinoma (RCC) is in part due to the hypermethylation of CpG islands in their putative promoter regions. Furthermore, we found that NELL1 suppresses and NELL2 partially suppresses RCC cell migration, and NELL1 further inhibits RCC cell adhesion. © 2015 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association

Quantitative analysis of β1,6GlcNAc-branched N-glycans on β4 integrin in cutaneous squamous cell carcinoma

α6β4 integrin plays pivotal roles in cancer progression in several types of cancers. Our previous study using N-glycan-manipulated cell lines demonstrated that defects in N-glycans or decreased β1,6GlcNAc-branched N-glycans on β4 integrin suppress β4 integrin-mediated cancer cell adhesion, migration, invasion, and tumorigenesis. Furthermore, immunohistochemical analysis has shown that colocalization of β1,6GlcNAc-branched N-glycans with β4 integrin was observed in cutaneous squamous cell carcinoma (SCC) tissue. However, until now there has been no direct evidence that β1,6GlcNAc-branched N-glycans are upregulated on β4 integrin in cutaneous SCC. In the present study, we performed an ELISA analysis of β1,6GlcNAc-branched N-glycans on β4 integrins as well as β4 integrins in cell lysates from human normal skin and cutaneous SCC tissues. The SCC samples showed a 4.9- to 7.4-fold increase in the ratio of β1,6GlcNAc-branched N-glycans to β4 integrin compared with normal skin samples. These findings suggest that the addition of β1,6GlcNAc-branched N-glycans onto β4 integrin was markedly elevated in cutaneous SCC tissue compared to normal skin tissue. The value of β1,6GlcNAc-branched N-glycans on β4 integrin may be useful as a diagnostic marker associated with cutaneous SCC tumor progression