EVL (Ena/VASP-like) expression is up-regulated in human breast cancer and its relative expression level is correlated with clinical stages.

EVL belongs to the Ena/VASP (Enabled/vasodilator-stimulated phosphoprotein) family of proteins [Mena (mammalian Ena), VASP and EVL], which have a range of roles in regulating the actin cytoskeleton. Growing evidence suggests that Mena and VASP are involved in carcinogenesis, though little is known about the significance of EVL's function in cancer. This study examined the expression levels of EVL mRNA in paired breast cancer specimens using semi-quantitative and real-time RT-PCR. In a comparison between matched breast tumors and normal tissues, a significant increase in the level of EVL mRNA was found in 23 of the 35 (65.7%) tumors (P=0.032). Patients in the advanced stages more frequently exhibited an elevated EVL expression in tumor tissues. The EVL mRNA relative expression level significantly correlated with clinical stages (P=0.021). To begin elucidating the mechanism, we measured the ability of EVL to transform NIH3T3 cells and regulate the motility of MCF-7 cells in vitro by focus formation and modified Boyden chamber assays. The results indicated that overexpression of EVL was insufficient to transform NIH3T3 cells, although the motility of EVL transfected MCF-7 cells was markedly promoted. Collectively, EVL expression level was higher in breast tumors compared to normal tissues and its up-regulation was positively associated with the clinical stages of breast cancer. Additionally, EVL may be implicated in invasion and/or metastasis of human breast cancer.

Biphasic expression of RhoGDI2 in the progression of breast cancer and its negative relation with lymph node metastasis.

RhoGDI2 has been shown to be a metastasis-related gene in several cancers. In human breast cancer, little clinical study of RhoGDI2 has been reported. In this study, we investigated the expression level of RhoGDI2 by immunohistochemistry, as well as the correlation of RhoGDI2 with clinicopathological parameters in 71 breast cancer specimens. We also examined RhoGDI2 expression at mRNA and protein levels of four human breast cancer cell lines differing in in vivo metastasis. Along with the extent of mammary epithelia proliferation and carcinogenesis, a biphasic pattern of RhoGDI2 expression (increase and then decrease) was observed, which was also found in these examined cells. Furthermore, univariate and multivariate analysis revealed that reduced expression of RhoGDI2 in the most malignant epithelia was significantly associated with lymph node metastasis (P<0.01). Our results suggest that RhoGDI2 may be implicated in the progress of malignancy and act as a metastasis-related marker in breast cancer.

Genome-wide ORFeome cloning and analysis of Arabidopsis transcription factor genes.

Here, we report our effort in generating an ORFeome collection for the Arabidopsis transcription factor (TF) genes. In total, ORFeome clones representing 1,282 Arabidopsis TF genes have been obtained in the Gateway high throughput cloning pENTR vector, including 411 genes whose annotation lack cDNA support. All the ORFeome inserts have also been mobilized into a yeast expression destination vector, with an estimated 85% rate of expressing the respective proteins. Sequence analysis of these clones revealed that 34 of them did not match with either the reported cDNAs or current predicted open-reading-frame sequences. Among those, novel alternative splicing of TF gene transcripts is responsible for the observed differences in at least five genes. However, those alternative splicing events do not appear to be differentially regulated among distinct Arabidopsis tissues examined. Lastly, expression of those TF genes in 17 distinct Arabidopsis organ types and the cultured cells was profiled using a 70-mer oligo microarray.