Gene-to-metabolite network for biosynthesis of lignans in MeJA-elicited Isatis indigotica hairy root cultures.

Root and leaf tissue of Isatis indigotica shows notable anti-viral efficacy, and are widely used as "Banlangen" and "Daqingye" in traditional Chinese medicine. The plants' pharmacological activity is attributed to phenylpropanoids, especially a group of lignan metabolites. However, the biosynthesis of lignans in I. indigotica remains opaque. This study describes the discovery and analysis of biosynthetic genes and AP2/ERF-type transcription factors involved in lignan biosynthesis in I. indigotica. MeJA treatment revealed differential expression of three genes involved in phenylpropanoid backbone biosynthesis (IiPAL, IiC4H, Ii4CL), five genes involved in lignan biosynthesis (IiCAD, IiC3H, IiCCR, IiDIR, and IiPLR), and 112 putative AP2/ERF transcription factors. In addition, four intermediates of lariciresinol biosynthesis were found to be induced. Based on these results, a canonical correlation analysis using Pearson's correlation coefficient was performed to construct gene-to-metabolite networks and identify putative key genes and rate-limiting reactions in lignan biosynthesis. Over-expression of IiC3H, identified as a key pathway gene, was used for metabolic engineering of I. indigotica hairy roots, and resulted in an increase in lariciresinol production. These findings illustrate the utility of canonical correlation analysis for the discovery and metabolic engineering of key metabolic genes in plants

Discrimination of Colon Cancer Stem Cells Using Noncanonical Amino Acid

Cancer stem cells (CSCs) may be responsible for tumor recurrence. Metabolic labelling of newly synthesized proteins with non-canonical amino acids allows us to discriminate CSCs in mixed populations due to the quiescent nature of these cells

Breast Tumour Initiating Cell Fate Is Regulated by Microenvironmental Cues from an Extracellular Matrix

Cancer stem cells, also known as tumour-initiating cells (TICs), are identified as highly tumorigenic population within tumours and hypothesized to be main regulators in tumour growth, metastasis and relapse. Evidence also suggests that a tumour microenvironment plays a critical role in the development and progression of cancer, by constantly modulating cell–matrix interactions. Scientists have tried to characterize and identify the TIC population but the actual combination of extracellular components in deciphering the fate of TICs has not been explored. The basic unanswered question is the phenotypic stability of this TIC population in a tissue extracellular matrix setting. The in vivo complexity makes it difficult to identify parameters in a diverse milieu that affect TICs behaviour. Herein we studied how the TIC population would respond when subjected to a unique microenvironment composed of different extracellularproteins. The TIC-enriched population isolated from a Her2/neu-induced mouse mammary tumour was cultured on collagen, fibronectin and laminin coated substrates for one to two weeks. Our observations indicate that a laminin substrate can maintain the majority of the self-renewing and tumorigenic TIC population, whereas collagen induced a more differentiated phenotype of the cells. Also interestingly, fibronectin substrates dictated an invasive phenotype of TICs as evidenced from the EMT-related gene expression pattern. The results of this study signify that the microenvironmental cues play a considerable role in tumour relapse and progression by altering the cancer stem cell behaviour and thus this knowledge could be used to design novel cancer therapeutics

The Effect of B27 Supplement on Promoting In Vitro Propagation of Her2/neu-Transformed Mammary Tumorspheres

A variety of human solid tumors, including breast cancer, are considered to embrace a hierarchical organization in which only rare tumor-initiating cancer stem cells are truly responsible for tumor formation. Cultivation of tumorspheres in non-adherent conditions is widely employed for enriching these putative cancer stem cells in vitro. However, the absence of a defined culture medium has handicapped further characterization and isolation of this cell population. In this study, we used mouse mammary tumor virus MMTV-HER2/neu transgenic mice that mimic the HER2/Neu-positive subtype of human breast cancer as a model system, and cultured primary tumor cells and tumorspheres from these mice under non-adherent conditions. In addition to essential growth factors, we found that B27 supplement played an important role in promoting tumorsphere formation and maintaining cultures through passaging. A tumorsphere-formation assay and measurements of average tumorsphere size provided insight into the characteristics of the putative breast cancer stem cells. Tumorspheres were enriched for cancer stem cells through serial passaging, while sphere size was determined by the innate properties of cancer stem cells and independent of the culture condition. Our study identifies B27 as an essential component of culture medium necessary for sustained propagation and enrichment of breast cancer stem cells in vitro

Expansion of Breast Cancer Stem Cells with Fibrous Scaffolds

Cancer stem cells (CSCs) are hypothesized as tumor-initiating cells within tumors and main contributors of tumor growth, metastasis and recurrence. Mammary cancer cells, MCF-7 cells, were cultured on 3D polycaprolactone (PCL) fibrous scaffolds, showing an increased proportion of CSCs. The expression of stem cell markers, including OCT3/4 and SOX2, and breast CSC-specific markers, SOX4 and CD49f, was significantly upregulated, and the mammosphere-forming capability in cells cultured on PCL fibrous scaffolds increased. The fibrous scaffolds also induced the elongation of MCF-7 cells and extended cell proliferation. The increase of CSC properties after being cultured on fibrous scaffolds was further confirmed with two luminal-type mammary cell lines, T47D and SK-BR-3, and a basal-type cell line, MDA-MB-231, by ALDEFLUOR assay and mammosphere formation assay. Moreover, we observed the upregulation of epithelial to mesenchymal transition and increased invasive capability in cells cultured on PCL fibrous scaffolds. These data suggest that the increase of CSC proportion in a 3D culture system may account for the enhanced malignancy. Therefore, our PCL fibrous scaffolds can potentially be used for CSCs enrichment and anti-cancer drug screening

A New Kind of Sub-pixel Interpolation Filtering Algorithm and Hardware Structural Design

Considering the problems of high complexity of sub-pixel interpolation operation and large visitor volume of storage in H.264/AVC standard, a kind of sub-pixel interpolation operation is put forward with changeable filter coefficient and unchangeable coefficient sum. According to the video image, filter coefficient can be confirmed. Based on the algorithm, a kind of 1/4 pixel precision interpolative hardware structural design. It is indicated by the performance analysis and filter structure that the structure is able to calculate pixel interpolation at different positions in a certain clock period with the characteristics of small area and fast speed. It is indicated by the result of experiment that compared with H.264 standard, new algorithm is able to reduce 18 % space complexity, improve noise ratio of peak, reduce bit rate and improve the performance of coding