ABSTRACT
Paclitaxel (trade name Taxol) is a rare diterpenoid with anticancer activity isolated from Taxus. At present, paclitaxel is mainly produced by the semi-synthetic method using extract of Taxus tissues as raw materials. The studies of regulatory mechanisms in paclitaxel biosynthesis would promote the production of paclitaxel through tissue/cell culture approaches. Here, we systematically identified 990 transcription factors (TFs), 460 microRNAs (miRNAs), and 160 phased small interfering RNAs (phasiRNAs) in Taxus chinensis to explore their interactions and potential roles in regulation of paclitaxel synthesis. The expression levels of enzyme genes in cone and root were higher than those in leaf and bark. Nearly all enzyme genes in the paclitaxel synthesis pathway were significantly up-regulated after jasmonate treatment, except for GGPPS and CoA Ligase. The expression level of enzyme genes located in the latter steps of the synthesis pathway was significantly higher in female barks than in male. Regulatory TFs were inferred through co-expression network analysis, resulting in the identification of TFs from diverse families including MYB and AP2. Genes with ADP binding and copper ion binding functions were overrepresented in targets of miRNA genes. The miRNA targets were mainly enriched with genes in plant hormone signal transduction, mRNA surveillance pathway, cell cycle and DNA replication. Genes in oxidoreductase activity, protein-disulfide reductase activity were enriched in targets of phasiRNAs. Regulatory networks were further constructed including components of enzyme genes, TFs, miRNAs, and phasiRNAs. The hierarchical regulation of paclitaxel production by miRNAs and phasiRNAs indicates a robust regulation at post-transcriptional level. Our study on transcriptional and posttranscriptional regulation of paclitaxel synthesis provides clues for enhancing paclitaxel production using synthetic biology technology.
ABSTRACT
Microorganisms are a unique part of our ecosystem because they affect the survival of living organisms. Although pathogenic microorganisms could be detrimental to the plants, animals, and humans, beneficial microbes have provided significant improvement in the growth and development of living organisms. In this study, the fungus Chaetomium globosium was isolated from the medicinal tree Gingko biloba, and then incorporated into a polymerization system to fabricate chitosan/acrylamide/gold (CS/Am/Au) nanocomposite hydrogels. The as-prepared hydrogel displayed increased mechanical strength due to the reinforcement of Au (gold) nanocomposites within the hydrogel matrix. Also, the equilibrium pH responsive swelling rates of the hydrogels gradually increased as the pH increases due to partial acid and basic hydrolysis occurring in the hydrogel as well as formation of hydrogen bond. In addition, the hydrogel demonstrated promising antibacterial activities against selected gram-positive (Staphylococcus epidermidis and Staphylococcus aureus) and gram-negative (Pseudomonas aeruginosa) bacterial strains with an average MIC90 of 0.125 mg/mL at a dosage of 1.0 mg/L. The obtained results are quite promising towards resolving several health challenges and advancing the pharmaceutical industries.
Subject(s)
Chaetomium , Chitosan , Nanocomposites , Animals , Humans , Chitosan/chemistry , Nanogels , Ginkgo biloba , Gold/pharmacology , Ecosystem , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Hydrogels/pharmacology , Hydrogels/chemistry , Nanocomposites/chemistry , AcrylamidesABSTRACT
Coronary artery ectasia (CAE) is a rare vascular phenotype characterized by abnormal dilation of blood vessels and disruption of coronary artery blood flow, which may promote thrombosis and an inflammatory response. We performed a cross-sectional study to investigate the association of white blood cells to mean platelet volume ratio (WMR) with CAE. Consecutive eligible patients (n = 492) were divided into two groups: including 238 patients with CAE and 254 patients in the normal coronary artery (NCA) group. WMR, the systemic immune-inflammation index (SII), and neutrophil-to-lymphocyte ratio (NLR) were found to be significantly associated with CAE in both univariate and multivariate logistic regression analyses. In multivariate analysis, the presence of WMR was associated with CAE (the odds ratios (OR) = 1.002, 95% CI: 1.001-1.003, P < .001). In the ROC analysis, the statistics (Z-values) of WMR vs SII and WMR vs NLR were 2.427 and 2.670 and were statistically significant (P = .015 and P = .008), indicating that WMR was superior to SII and NLR in distinguishing WMR. The optimal cut-off value was calculated from the point of maximal sensitivity and specificity by using Youden's index, which was determined to be 635.50. WMR has the potential to be a cost-effective tool to monitor CAE.
ABSTRACT
Background: This study aimed to develop an artificial intelligence-based computer-aided diagnosis system (AI-CAD) emulating the diagnostic logic of radiologists for lymph node metastasis (LNM) in esophageal squamous cell carcinoma (ESCC) patients, which contributed to clinical treatment decision-making. Methods: A total of 689 ESCC patients with PET/CT images were enrolled from three hospitals and divided into a training cohort and two external validation cohorts. 452 CT images from three publicly available datasets were also included for pretraining the model. Anatomic information from CT images was first obtained automatically using a U-Net-based multi-organ segmentation model, and metabolic information from PET images was subsequently extracted using a gradient-based approach. AI-CAD was developed in the training cohort and externally validated in two validation cohorts. Results: The AI-CAD achieved an accuracy of 0.744 for predicting pathological LNM in the external cohort and a good agreement with a human expert in two external validation cohorts (kappa = 0.674 and 0.587, p < 0.001). With the aid of AI-CAD, the human expert's diagnostic performance for LNM was significantly improved (accuracy [95% confidence interval]: 0.712 [0.669-0.758] vs. 0.833 [0.797-0.865], specificity [95% confidence interval]: 0.697 [0.636-0.753] vs. 0.891 [0.851-0.928]; p < 0.001) among patients underwent lymphadenectomy in the external validation cohorts. Conclusions: The AI-CAD could aid in preoperative diagnosis of LNM in ESCC patients and thereby support clinical treatment decision-making.
ABSTRACT
Centromeres are long, often repetitive regions of genomes that bind kinetochore proteins and ensure normal chromosome segregation. Engineering centromeres that function in vivo has proven to be difficult. Here we describe a tethering approach that activates functional maize centromeres at synthetic sequence arrays. A LexA-CENH3 fusion protein was used to recruit native Centromeric Histone H3 (CENH3) to long arrays of LexO repeats on a chromosome arm. Newly recruited CENH3 was sufficient to organize functional kinetochores that caused chromosome breakage, releasing chromosome fragments that were passed through meiosis and into progeny. Several fragments formed independent neochromosomes with centromeres localized over the LexO repeat arrays. The new centromeres were self-sustaining and transmitted neochromosomes to subsequent generations in the absence of the LexA-CENH3 activator. Our results demonstrate the feasibility of using synthetic centromeres for karyotype engineering applications.
Subject(s)
Centromere , Zea mays , Zea mays/genetics , Zea mays/metabolism , Centromere/genetics , Kinetochores/metabolism , Histones/metabolism , Cell CycleABSTRACT
Demethylation of transposons can activate the expression of nearby genes and cause imprinted gene expression in the endosperm; this demethylation is hypothesized to lead to expression of transposon small interfering RNAs (siRNAs) that reinforce silencing in the next generation through transfer either into egg or embryo. Here we describe maize (Zea mays) maternal derepression of r1 (mdr1), which encodes a DNA glycosylase with homology to Arabidopsis thaliana DEMETER and which is partially responsible for demethylation of thousands of regions in endosperm. Instead of promoting siRNA expression in endosperm, MDR1 activity inhibits it. Methylation of most repetitive DNA elements in endosperm is not significantly affected by MDR1, with an exception of Helitrons. While maternally-expressed imprinted genes preferentially overlap with MDR1 demethylated regions, the majority of genes that overlap demethylated regions are not imprinted. Double mutant megagametophytes lacking both MDR1 and its close homolog DNG102 result in early seed failure, and double mutant microgametophytes fail pre-fertilization. These data establish DNA demethylation by glycosylases as essential in maize endosperm and pollen and suggest that neither transposon repression nor genomic imprinting is its main function in endosperm.
Subject(s)
Arabidopsis , DNA Glycosylases , Arabidopsis/genetics , DNA/metabolism , DNA Glycosylases/genetics , DNA Glycosylases/metabolism , DNA Methylation/genetics , Endosperm/genetics , Endosperm/metabolism , Gene Expression Regulation, Plant/genetics , Genomic Imprinting/genetics , RNA, Small Interfering/genetics , Zea mays/genetics , Zea mays/metabolismABSTRACT
Ginkgo (Ginkgo biloba L.) is a deciduous tree species with high timber, medicinal, ecological, ornamental, and scientific values, and is widely cultivated worldwide. However, for such an important tree species, the regulatory mechanisms involved in the photosynthesis of developing leaves remain largely unknown. Here, we observed variations in light response curves (LRCs) and photosynthetic parameters (photosynthetic capacity (Pnmax) and dark respiration rate (Rd)) of leaves across different developmental stages. We found the divergence in the abundance of compounds (such as 3-phospho-d-glyceroyl phosphate, sedoheptulose-1,7-bisphosphate, and malate) involved in photosynthetic carbon metabolism. Additionally, a co-expression network was constructed to reveal 242 correlations between transcription factors (TFs) and photosynthesis-related genes (p < 0.05, |r| > 0.8). We found that the genes involved in the photosynthetic light reaction pathway were regulated by multiple TFs, such as bHLH, WRKY, ARF, IDD, and TFIIIA. Our analysis allowed the identification of candidate genes that most likely regulate photosynthesis, primary carbon metabolism, and plant development and as such, provide a theoretical basis for improving the photosynthetic capacity and yield of ginkgo trees.
Subject(s)
Ginkgo biloba/genetics , Metabolome/genetics , Photosynthesis/genetics , Transcriptome/genetics , Gene Expression Profiling/methods , Plant Leaves/genetics , Plant Proteins/genetics , Transcription Factors/geneticsABSTRACT
As one of the largest families of transcription factors, the R2R3-MYB family plays a significant role in plant growth, development, and response to hormone and environmental stress. To explore its evolutionary mechanism and potential function in Ginkgo biloba, a gymnosperm of great economic and ecological value, we presented a comprehensive analysis of the R2R3-MYB genes in ginkgo. Sixty-nine GbR2R3-MYB genes were identified and these genes could be classified into 33 groups based on the characteristics of the amino acid sequence of the R2R3-MYB domain and gene structure. Syntenic analyses indicated that few tandem and segmental duplications possibly resulted in the contraction of the GbR2R3-MYB gene family. Based on the transcriptome data, expression profiles of eight different tissues and different developmental stages of leaf and kernel showed that GbR2R3-MYB genes had distinct temporal and spatial expression characteristics. Specific expression patterns of the sixteen GbR2R3-MYB genes were also identified in response to different abiotic stresses and hormonal exposures. Further investigation revealed that GbR2R3-MYB19 was located in the nucleus and possessed transcriptional activity, implying its potential roles in the regulation of multiple biological processes. Our findings provide a robust basis for future comprehensive evolutionary and functional analyses of GbR2R3-MYB genes in ginkgo.
Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Gene Expression Regulation, Plant , Ginkgo biloba/genetics , Transcription Factors/genetics , Transcriptome , Amino Acid Sequence , Arabidopsis/classification , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Cell Nucleus/genetics , Cell Nucleus/metabolism , Gene Expression Regulation, Developmental , Ginkgo biloba/classification , Ginkgo biloba/growth & development , Ginkgo biloba/metabolism , Multigene Family , Phylogeny , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Leaves/metabolism , Protein Isoforms/classification , Protein Isoforms/genetics , Protein Isoforms/metabolism , Seeds/genetics , Seeds/growth & development , Seeds/metabolism , Sequence Alignment , Stress, Physiological/genetics , Transcription Factors/metabolismABSTRACT
Ginkgo (Ginkgo biloba L.) is a high-value medicinal tree species characterized by its flavonoids beneficial effects that are abundant in leaves. We performed a temporospatial comprehensive transcriptome and metabolome dynamics analyses of clonally propagated Ginkgo plants at four developmental stages (time: May to August) across three different environments (space) to unravel leaves flavonoids biosynthesis variation. Principal component analysis revealed clear gene expression separation across samples from different environments and leaf-developmental stages. We found that flavonoid-related metabolism was more active in the early stage of leaf development, and the content of total flavonoid glycosides and the expression of some genes in flavonoid biosynthesis pathway peaked in May. We also constructed a co-expression regulation network and identified eight GbMYBs and combining with other TF genes (3 GbERFs, 1 GbbHLH, and 1 GbTrihelix) positively regulated the expression of multiple structural genes in the flavonoid biosynthesis pathway. We found that part of these GbTFs (Gb_11316, Gb_32143, and Gb_00128) expressions was negatively correlated with mean minimum temperature and mean relative humidity, while positively correlated with sunshine duration. This study increased our understanding of the molecular mechanisms of flavonoids biosynthesis in Ginkgo leaves and provided insight into the proper production and management of Ginkgo commercial plantations.
ABSTRACT
Colletotrichum gloeosporioides is a hemibiotrophic pathogen causing significant losses to economically important crops and forest trees, including Liriodendron. To explore the interaction between C. gloeosporioides and Liriodendron and to identify the candidate genes determining the pathogenesis, we sequenced and assembled the whole genome of C. gloeosporioides Lc1 (CgLc1) using PacBio and Illumina next generation sequencing and performed a comparative genomic analysis between CgLc1 and Cg01, the latter being a described endophytic species of the C. gloeosporioides complex. Gene structure prediction identified 15,744 protein-coding genes and 837 noncoding RNAs. Species-specific genes were characterized using an ortholog analysis followed by a pathway enrichment analysis, which showed that genes specific to CgLc1 were enriched for the arginine biosynthetic process. Furthermore, genome synteny analysis revealed that most of the protein-coding genes fell into collinear blocks. However, two clusters of polyketide synthase genes were identified to be specific for CgLc1, suggesting that they might have an important role in virulence control. Transcriptional regulators coexpressed with polyketide synthase genes were detected through a Weighted Correlation Network Analysis. Taken together, this work provides new insight into the virulence- and pathogenesis-associated genes present in C. gloeosporioides and its possible lifestyle.
Subject(s)
Colletotrichum , Liriodendron , Plant Diseases , Plant Leaves , VirulenceABSTRACT
Gametes constitute a critical stage of the plant life cycle during which the genome undergoes reprogramming in preparation for embryogenesis. Here, we examined genome-wide distributions of small RNAs in the sperm and egg cells of rice. We found that 24-nt siRNAs, which are a hallmark of RNA-directed DNA methylation (RdDM) in plants, were depleted from heterochromatin boundaries in both gametes relative to vegetative tissues, reminiscent of siRNA patterns in DDM1-type nucleosome remodeler mutants. In sperm cells, 24-nt siRNAs were spread across heterochromatic regions, while in egg cells, 24-nt siRNAs were concentrated at a smaller number of heterochromatic loci throughout the genome, especially at loci which also produced siRNAs in other tissues. In both gametes, patterns of CHH methylation, typically a strong indicator of RdDM, were similar to vegetative tissues, although lower in magnitude. These findings indicate that the small RNA transcriptome undergoes large-scale redistribution in both male and female gametes, which is not correlated with recruitment of DNA methyltransferases in gametes and suggestive of unexplored regulatory activities of gamete small RNAs.
Subject(s)
Germ Cells/growth & development , Oryza/genetics , RNA, Small Interfering/genetics , Sex Determination Processes/genetics , DNA Methylation/genetics , Gene Expression Regulation, Plant/genetics , Gene Silencing , Genome, Plant/genetics , Heterochromatin/genetics , Nucleosomes/genetics , Oryza/growth & development , Transcriptome/geneticsABSTRACT
Biolistic transformation delivers nucleic acids into plant cells by bombarding the cells with microprojectiles, which are micron-scale, typically gold particles. Despite the wide use of this technique, little is known about its effect on the cell's genome. We biolistically transformed linear 48-kb phage lambda and two different circular plasmids into rice (Oryza sativa) and maize (Zea mays) and analyzed the results by whole genome sequencing and optical mapping. Although some transgenic events showed simple insertions, others showed extreme genome damage in the form of chromosome truncations, large deletions, partial trisomy, and evidence of chromothripsis and breakage-fusion bridge cycling. Several transgenic events contained megabase-scale arrays of introduced DNA mixed with genomic fragments assembled by nonhomologous or microhomology-mediated joining. Damaged regions of the genome, assayed by the presence of small fragments displaced elsewhere, were often repaired without a trace, presumably by homology-dependent repair (HDR). The results suggest a model whereby successful biolistic transformation relies on a combination of end joining to insert foreign DNA and HDR to repair collateral damage caused by the microprojectiles. The differing levels of genome damage observed among transgenic events may reflect the stage of the cell cycle and the availability of templates for HDR.
Subject(s)
DNA, Plant/genetics , Genome, Plant/genetics , Oryza/genetics , Zea mays/genetics , BiolisticsABSTRACT
Plants make use of distinct types of DNA methylation characterized by their DNA methyltransferases and modes of regulation. One type, RNA-directed DNA methylation (RdDM), is guided by small interfering RNAs (siRNAs) to the edges of transposons that are close to genes, areas called mCHH islands in maize (Zea mays). Another type, chromomethylation, is guided by histone H3 lysine 9 methylation to heterochromatin across the genome. We examined DNA methylation and small RNA expression in plant tissues that were mutant for both copies of the genes encoding chromomethylases as well as mutants for both copies of the genes encoding DECREASED DNA METHYLATION1 (DDM1)-type nucleosome remodelers, which facilitate chromomethylation. Both sets of double mutants were nonviable but produced embryos and endosperm. RdDM was severely compromised in the double mutant embryos, both in terms of DNA methylation and siRNAs. Loss of 24-nucleotide siRNA from mCHH islands was coupled with a gain of 21-, 22-, and 24-nucleotide siRNAs in heterochromatin. These results reveal a requirement for both chromomethylation and DDM1-type nucleosome remodeling for RdDM in mCHH islands, which we hypothesize is due to dilution of RdDM components across the genome when heterochromatin is compromised.
Subject(s)
DNA (Cytosine-5-)-Methyltransferases/metabolism , Gene Expression Regulation, Plant/physiology , Plant Proteins/metabolism , Zea mays/metabolism , DNA (Cytosine-5-)-Methyltransferases/genetics , DNA Methylation/genetics , DNA Methylation/physiology , Gene Expression Regulation, Plant/genetics , Mutation/genetics , Plant Proteins/genetics , Zea mays/geneticsABSTRACT
The midkine antisense oligonucleotide (MK-ASODN, 5'-CCC CGG GCC GCC CTT CTT CA-3') nanoliposomes have been identified to suppress hepatocellular carcinoma (HCC) growth effectively, and have a great potential to be an effective target drug for HCC. In this study, a facile and reproducible method for large-scale preparation of MK-ASODN nanoliposomes followed by lyophilization has been developed successfully. Meanwhile, the MK-ASODN nanoliposomes characteristics, storage stability and their antitumor efficiency were studied. The mean particle size of MK-ASODN nanoliposomes were 229.43±15.11 nm, and the zeta potential were 29.7±1.1 mV. High entrapment efficiency values were achieved around 90%. Transmission electron microscopy images revealed spherical shaped nanoliposomes. Nanoliposomes allowed sustained MK-ASODN release for as long as 14 days. During 180 days of storage, freeze-dried nanoliposomes showed no significant change in the mean size, zeta potential, entrapment efficiency and drug release ratio. Regarding their antitumor efficiency, the in vitro proliferation of human liver cancer cells were significantly inhibited by the MK-ASODN nanoliposomes. Furthermore, the MK-ASOND nanoliposomes also significantly inhibited the growth of HCC in the mouse model. In summary, the results confirmed that this large-scale preparation of MK-ASOND nanoliposomes was facile and reproducible, and potentially, could speed up the application process of our MK-ASOND nanoliposomes for HCC therapy.
Subject(s)
Carcinoma, Hepatocellular/drug therapy , Cytokines/genetics , Liver Neoplasms/drug therapy , Oligonucleotides, Antisense/administration & dosage , Animals , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Drug Compounding/methods , Drug Stability , Drug Storage , Female , Freeze Drying , Humans , Liposomes , Liver Neoplasms/pathology , Male , Mice , Mice, Inbred BALB C , Microscopy, Electron, Transmission , Midkine , Nanoparticles , Oligonucleotides, Antisense/pharmacology , Particle Size , Pressure , Reproducibility of Results , UltrafiltrationABSTRACT
Colorectal cancer remains one of the most common types of cancer and leading causes of cancer death worldwide. Although we have made steady progress in chemotherapy and targeted therapy, evidence suggests that the majority of patients undergoing drug therapy experience severe, debilitating, and even lethal adverse drug events which considerably outweigh the benefits. The identification of suitable biomarkers will allow clinicians to deliver the most appropriate drugs to specific patients and spare them ineffective and expensive treatments. Prognostic and predictive biomarkers have been the subjects of many published papers, but few have been widely incorporated into clinical practice. Here, we want to review recent biomarker data related to colorectal cancer, which may have been ready for clinical use.
Subject(s)
Biomarkers, Tumor/genetics , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/genetics , Antineoplastic Agents/adverse effects , Antineoplastic Agents/metabolism , Camptothecin/adverse effects , Camptothecin/analogs & derivatives , Camptothecin/metabolism , Chemotherapy, Adjuvant/adverse effects , Colorectal Neoplasms/metabolism , ErbB Receptors/antagonists & inhibitors , Fluorouracil/adverse effects , Fluorouracil/metabolism , Humans , Irinotecan , Molecular Targeted Therapy , Organoplatinum Compounds/adverse effects , Organoplatinum Compounds/metabolism , Oxaliplatin , Predictive Value of Tests , Prognosis , Transcriptome , Vascular Endothelial Growth Factor A/antagonists & inhibitorsABSTRACT
Starch biosynthesis is important for plant development and is a critical factor in crop quality and nutrition. As a complex metabolic pathway, the regulation of starch biosynthesis is still poorly understood. We here present the identification of candidate regulators for starch biosynthesis by gene coexpression analysis in rice (Oryza sativa). Starch synthesis genes can be grouped into type I (in seeds; sink tissues) and type II (in vegetative tissues; source tissues), and 307 and 621 coexpressed genes are putatively involved in the regulation of starch biosynthesis in rice seeds and vegetative tissues, respectively. Among these genes, Rice Starch Regulator1 (RSR1), an APETALA2/ethylene-responsive element binding protein family transcription factor, was found to negatively regulate the expression of type I starch synthesis genes, and RSR1 deficiency results in the enhanced expression of starch synthesis genes in seeds. Seeds of the knockout mutant rsr1 consistently show the increased amylose content and altered fine structure of amylopectin and consequently form the round and loosely packed starch granules, resulting in decreased gelatinization temperature. In addition, rsr1 mutants have a larger seed size and increased seed mass and yield. In contrast, RSR1 overexpression suppresses the expression of starch synthesis genes, resulting in altered amylopectin structure and increased gelatinization temperature. Interestingly, a decreased proportion of A chains in rsr1 results in abnormal starch granules but reduced gelatinization temperature, whereas an increased proportion of A chains in RSR1-overexpressing plants leads to higher gelatinization temperatures, which is novel and different from previous reports, further indicating the complicated regulation of starch synthesis and determination of the physicochemical properties of starch. These results demonstrate the potential of coexpression analysis for studying rice starch biosynthesis and the regulation of a complex metabolic pathway and provide informative clues, including the characterization of RSR1, to facilitate the improvement of rice quality and nutrition.
Subject(s)
DNA-Binding Proteins/metabolism , Oryza/genetics , Plant Proteins/metabolism , Starch/biosynthesis , Amylopectin/chemistry , DNA, Bacterial/genetics , DNA-Binding Proteins/genetics , Endosperm/chemistry , Gene Expression Regulation, Plant , Molecular Sequence Data , Mutagenesis, Insertional , Oligonucleotide Array Sequence Analysis , Oryza/metabolism , Plant Proteins/genetics , RNA, Plant/genetics , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
AIM: To synthesize antisense oligonucleotides (ASODNs) of midkine (MK), package the ASODNs with nanoparticles, and to inhibit hepatocellular carcinoma (HCC) growth using these nanoparticles. METHODS: HepG2 cell proliferation was analyzed in vitro using the 3-(4,5-dimethythiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt assay. The in vivo activity of nanoparticles delivering the MK-ASODNs was analyzed by histopathological and immunohistochemical staining and quantitative real time polymerase chain reaction (PCR). RESULTS: The in vitro proliferation of HepG2 cells was significantly inhibited by the nanoparticles packaged with MK-ASODNs (NANO-ASODNs). Furthermore, the NANO-ASODNs significantly inhibited the growth of HCC in the mouse model. CONCLUSION: NANO-ASODNs can significantly suppress the growth of HCC in vitro and in vivo.
Subject(s)
Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/pathology , Cytokines/therapeutic use , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Nanoparticles , Oligonucleotides, Antisense/therapeutic use , Animals , Biomarkers, Tumor/metabolism , Carcinoma, Hepatocellular/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cytokines/genetics , Female , Humans , Liver Neoplasms/metabolism , Mice , Mice, Inbred BALB C , Mice, Nude , Midkine , Molecular Sequence Data , Nanoparticles/chemistry , Nanoparticles/therapeutic use , Oligonucleotides, Antisense/genetics , Random Allocation , alpha-Fetoproteins/metabolismABSTRACT
A rice (Oryza sativa) T-DNA insertion population, which included more than 63 000 independent transgenic lines and 8 840 identified flanking sequence tags (FSTs) that were mapped onto the rice genome, was developed to systemically study the rice seed quality control. Genome-wide analysis of the FST distribution showed that T-DNA insertions were positively correlated with expressed genes, but negatively with transposable elements and small RNAs. In addition, the recovered T-DNAs were preferentially located at the untranslated region of the expressed genes. More than 11 000 putative homozygous lines were obtained through multi-generations of planting and resistance screening, and measurement of seed quality of around half of them, including the contents of starch, amylose, protein and fat, with a nondestructive near-infrared spectroscopy method, identified 551 mutants with unique or multiple altered parameters of seed quality. Analysis of the corresponding FSTs showed that genes participating in diverse functions, including metabolic processes and transcriptional regulation, were involved, indicating that seed quality is regulated by a complex network.
Subject(s)
DNA, Bacterial/genetics , Mutagenesis, Insertional , Oryza/genetics , Seeds/genetics , Chromosomes, Plant/genetics , DNA, Intergenic/genetics , Genes, Plant , Genetic Linkage , Homozygote , Mutation/genetics , Spectroscopy, Near-InfraredABSTRACT
To study knockdown effect of small interfering RNA (siRNA) to PLK1 (Polo-like kinase 1) mRNA in colorectal cancer cell line SW480 and its mitosis and growth was changed. Ten special siRNA molecules were designed targeting different sites of PLK1 mRNA sequence and chemically synthesized. The siRNA molecules were transfected into SW480 by Oligofectamine. The gene mRNA level was assayed by Real-Time PCR. The changes of PLK1 protein, SW480 cell cycle and survival percentage was checked by Western-blot, Flow cytometry and Cell counter assays respectively. All 10 siRNA molecules knocked PLK1 mRNA down in different level. Of them P1, P4 and P9 showed over 80% knockdown efficiency and the others had more than 20% knockdown efficiency to PLK1 mRNA. The best knockdown effect over 95% of all groups was at 25 nmol/L of a mixture with P1, P4 and P9 siRNA equally. In this situation the protein was very less and the cells were blocked at G2 phase of cell cycle. After 72 h cell survival percentages were consistent with PKL1 mRNA level change by siRNA gradient concentration. The results showed that siRNA targeting PLK1 mRNA had effectively knocked PLK1 mRNA down in SW480 cell line. And a blended siRNAs held the best knockdown effect. The cell was blocked on the mitosis and growth.