TARGETING OF CANCER STEM CELLS BY LET-7 miRNA

miRNAs are short non-coding RNA that regulate gene expression at the post-transcriptional level by inhibiting the translation of mRNAs through pairing to their 3’ UTR. The let-7 family of miRNAs regulates cell differentiation during embryogenesis and is responsible for the maintenance of the differentiated state in adult cells. Let-7 miRNA levels are often reduced in malignant tumors and its ectopic expression in cancer cells causes cell growth arrest, reduces invasiveness and down-regulates several oncogenes. To gain a further understanding of their biological functions, the breast cancer cell line MDA-MB-231 was transfected with let-7 miRNA mimics and their effect was examined by qPCR, western blot, flow cytometry and functional assays to determine stem cell characteristics and differentiation. We confirmed that let-7 miRNA mimics reduced cell proliferation, and HMGA2, Cyclin D1, Ras and Lin28A protein level in MDA-MB-231 cells. We found that let-7 miRNA down-regulated the levels of active β-catenin (ABC). We showed for the first time that let-7 miRNAs coordinately induce the enzymes in the serine biosynthesis pathway at the transcriptional level in MDA-MB-231 cells. Furthermore, we found that the protein level of the enzymes is differentially regulated: whereas the first two enzymes of the pathway are up-regulated, the last enzyme, phosphoserine phosphatase (PSPH), is down-regulated. ABC down-regulation by let-7 mimics is consistent with the tumor suppressor role of let-7 miRNA family and represents an additional mechanism by which let-7 interferes with Wnt pathway. The regulation of serine biosynthesis pathway is a novel and unexpected function of let-7 that raises many questions and leads to the exciting emerging field: Metaboloepigenetics

Evaluation of Image Warping Algorithms for Implementation in FPGA

The target of this master thesis is to evaluate the Image Warping technique and propose a possible design for an implementation in FPGA. The Image Warping is widely used in the image processing for image correction and rectification. A DSP is a usual choice for implantation of the image processing algorithms, but to decrease a cost of the target system it was proposed to use an FPGA for implementation. In this work a different Image Warping methods was evaluated in terms of performance, produced image quality, complexity and design size. Also, considering that it is not only Image Warping algorithm which will be implemented on the target system, it was important to estimate a possible memory bandwidth used by the proposed design. The evaluation was done by implemented a C-model of the proposed design with a finite datapath to simulate hardware implementation as close as possible

Additional file 1: of Real-Time Vital Mineralization Detection and Quantification during In Vitro Osteoblast Differentiation

Figure S1. a An example of the mask set for the green objects count on day 10; left pictures shows green fluorescent dots corresponding to the forming calcium crystals; right pictures shows how the IncuZOOM mask depicted green dots with high sensitivity (even very weak and small green dots are counted), representative pictures are shown. b Real-time calcium crystals formation detected by fluorescent imaging in calcein-treated and calcein-untreated samples, n = 3, SE. c Calcein detection in negative control and PPP2R2C siRNA treated samples, representative pictures are shown. d Number of dead cells detected by propidium iodide in calcein-treated and calcein untreated cells, n = 2, SE. (TIF 2900 kb

Accurate 3-gene-signature for early diagnosis of liposarcoma progression

Background Well- and dedifferentiated liposarcoma (WD/DDLPS) are rare mesenchymal malignant tumors that account for 20% of all sarcomas in adults. The WD form is a low-grade malignancy with a favourable prognosis which may progress to DDLPS, a high-grade aggressive counterpart. WDLPS is referred to as atypical lipomatous tumour (ALT) when localised in extremities, due to its better prognosis. Currently the final differential diagnosis to distinguish between more aggressive and less aggressive form is based on post-surgical histological examination and no molecular biomarkers for early detection are available. Methods Quantitative polymerase chain reaction (qPCR) analysis of 11 metabolic genes involved in general and adipose tissue-specific metabolism, was performed on ALT (= 8), WDLPS (= 9) and DDLPS (= 20) samples. Subsequent statistical analysis was carried out to determine genes that most accurately can predict DDLPS differential diagnosis. Selected genes were further validated in a separate cohort by qPCR and the data statistically analysed. Deep sequencing was performed on DDLPS specimen from the metastatic patient and on five random WDLPS specimens. Results We established a three-gene signature based on PNPLA2, LIPE and PLIN1, which identified DDLPS with 100% sensitivity and 90% specificity, even in specimens from the WD component of DDLPS tumors. Interestingly, the PNPLA2 gene is deleted in 45% of DDLPS samples analyzed under TCGA project, and the deletion is associated with significantly lower PNPLA2 expression level. However, other mechanisms causing loss or downregulation of the expression of these three genes may be involved. Moreover, the significantly lower level of PNPLA2 is associated with R1 surgical margins, compare to R0 margins, which suggests the more invasive tumor phenotype in the absence of PNPLA2. Conclusions The identified metabolic signature allows highly accurate differential diagnosis between WD- and DDLPS even in samples containing lipid droplets, a marker of differentiation, which makes it very suitable for the use on biopsies. In respect to the pathogenesis of the disease, our results give a new insight into possible molecular mechanisms involved and support the recent observation that deletion of PNPLA2 is a novel factor in liposarcoma progression