CENPU Promotes Growth and Metastasis in Nasopharyngeal Carcinoma by Inhibiting DUSP6

Background: Centromere protein U (CENPU), a centromere component, is key for mitosis and involved in the carcinogenesis of cancers. The role and mechanisms of CENPU in nasopharyngeal carcinoma (NPC) has not been described. Methods: CENPU expression in NPC cells and tissues was evaluated by RT-PCR and western blotting. Clinical significance of CENPU was evaluated by Immunohistochemistry. Biological functions of CENPU were evaluated by cell growth assay, colony formation assay, apoptosis assay, migration assay and invasion assays. Xenograft growth and lung metastasis model were conducted to investigate the effect of CENPU in vivo. Gene chip analysis, Ingenuity Pathway Analysis (IPA), and co-immunoprecipitation (Co-IP) experiments were used to explore the mechanisms of CENPU in NPC.Results: CENPU was highly expressed in NPC cells and samples. Patients with CENPU positive expression were closely associated with poor overall survival. Knockdown of CENPU inhibited proliferation and migration in vitro and in vivo in NPC. Gene chip analysis and IPA suggested that differentially expressed genes (DEGs) were significantly enriched in cancer and functions, including cellular movement, cellular development, cell growth and death, and proliferation when CENPU was downregulated. Dual specificity phosphatase 6 (DUSP6) was one of the DEGs and significantly decreased in NPC samples, and inversely correlated with expression with CENPU. Mechanism studies confirmed that CENPU increased the activation of ERK1/2 and p38 signal pathways by suppressing the expression of DUSP6. Therefore, our results suggested that CENPU might act as an oncogene in NPC and promote the development of NPC via inhibition of DUSP6, resulting in the inactivation of Erk1/2 and p38 pathways. Conclusions: CENPU facilitated cell proliferation and invasion by interacting with DUSP6. CENPU might be a promising prognostic biomarker and a potential target for NPC therapy.

Analysis of the application of a gene chip method for detecting Mycobacterium tuberculosis drug resistance in clinical specimens: a retrospective study

Most Mycobacterium tuberculosis (Mtb) resistant to rifampicin (RIF) has mutations in the rpoB gene, while most Mtb resistant to isoniazid (INH) has mutations in the katG gene or inhA promoter. We used gene chip technology to detect mutations in these genes to determine the resistance of Mtb to RIF and INH. A total of 4148 clinical specimens with sputum smear positivity for acid-fast bacilli (AFB) were detected. Then, taking the results of the drug sensitivity test (DST) as the reference standard, the detection efficiency of sputum samples from different grades of positive smears was compared in detail. We found that the sensitivity of the gene chip method for detecting sputum samples with a grade ≥ AFB 2 + was higher than that of sputum samples with a grade ≤ AFB 1 + (P < 0.05). When the grade of the sample was ≤ AFB 1 +, the sensitivity of the gene chip method was 72.6% for RIF, 67.3% for INH, and 60.0% for MDR-TB. When the grade of the sample was ≥ AFB 2 +, the sensitivity of the gene chip method was 84.5% for RIF, 78.2% for INH, and 73.9% for MDR-TB. The results show that gene chip technology can be directly used to diagnose drug-resistant tuberculosis in clinical specimens, and the diagnostic efficiency for the detection of sputum specimens with a grade ≥ AFB 2 + is better than that of other sputum specimens.

MYOSTATIN COORDINATING THE PROLIFERATION AND DIFFERENTIATION OF ADIPOSE AND SKELETAL MUSCLE CELLS AND ENERGY METABOLISM BALANCE BASED ON GENE CHIP TECHNOLOGY

ABSTRACT Myoblasts fuse into multinucleated muscle fibers to form and promote the growth of skeletal muscle. In order to analyze the role of myostatin (MSTN) in body fat, skeletal muscle cell proliferation and differentiation and energy metabolism, this study will use the antisense RNA technology of gene chip technology to study it. The results showed that the MSTN gene regulated the growth and proliferation of myoblasts and affected the development of skeletal muscle by affecting the expression of Cdc42, bnip2, p38 and other genes; knockout or overexpression of the MSTN gene would lead to a trend of fat-related genes from fat synthesis to fat decomposition; after the MSTN gene was knocked down, the expression levels of cpti-b, PPARG and other genes in the cells were corresponding after MSTN overexpression, the relative expression of the PPARG gene decreased. It is suggested that the knockout or overexpression of MSTN may affect lipid accumulation, and cpti-b and PPARG may directly regulate lipid level. It is hoped that this experiment can provide a reference for the study of MSTN effect on fat deposition.

Data Mining Analysis of Gene Prognostic Markers of Metastatic Skin Cancer Based on the Elastic Network Method

Skin cancer is a typical cancer tumor, which occurs all over the world and has a relatively high recurrence rate, including metastatic tumors that occur in other tissues and metastases to the skin, thus jeopardizing the personal life satisfaction and soundness of patients. Due to individual differences, the traditional treatment methods cannot adapt to every patient accurately, so it is difficult to achieve the desired treatment effect for each individual. Nowadays, with the development of gene chip, many new therapies based on gene are more targeted and flexible for the treatment of skin cancer patients. Therefore, it is necessary to mine and analyze appropriate gene biomarkers according to patients' genes. Because of the high cost of gene chip technology and the large number of human genes, there are few samples of gene data and high dimensions. It is a key problem to mine effective genetic biomarkers from the sample data. In this paper, we firstly performed the preliminary analysis using the difference expression analysis and proportional hazards model, then used the elastic network method to reduce the range of genetic data selection, and screened 26 gene prognostic markers closely related to the recurrence of metastatic skin cancer. Finally, the 26 gene biomarkers were analyzed by functional analysis and verified using a test sample. Research findings have shown that the obtained genetic markers have certain value in the clinical prognostic treatment of metastatic skin cancer.

The accumulation of exosome-associated microRNA-1246 and microRNA-150-3p in human red blood cell suspensions

Background Transfusion-related immunomodulation (TRIM) can be caused by exosomes, in which case, microRNAs (miRNAs) are one critical factor impacting exosome behavior. This study aims to investigate and analyze the expression profiles of exosomal miRNA in red blood cell (RBC) suspensions during storage and to identify potential TRIM-related miRNAs as well as their potential functions. Methods A total of 25 packs of RBC suspensions were randomly collected. Exosome were extracted by ultracentrifugation and then identified and characterized by nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM) and western blot (WB). Exosomal miRNA profiles were acquired using gene chips in five packs on week 1 and week 5. The expression data were compared from the two time points identifying accumulated miRNAs with statistical significance and their predicted targeting genes were analyzed. Based on the gene chip results, quantitative reverse transcription-polymerase chain reactions (qRT-PCR) were performed to verify miRNA accumulation in the rest 20 packs sampling on week 1, 3 and 5. Results Gene chip analysis revealed that most exosomal miRNAs were enriched as the storage period progressed. Compared to samples from week 1, week 5 samples exhibited a total of 539 differential miRNA expressions, among which, 159 were statistically significant (P < 0.05) and 148 (93.08%) were accumulated. In the bioinformatics functional analysis, significant immunoregulatory annotations related to the thyroid hormone, mitogen-activated protein kinase (MAPK), focal adhesion and RAS signaling pathways were identified. The top 17 differential expression miRNAs were validated by qRT-PCR. The results confirmed that all the 17 miRNAs were accumulated with increasing storage time. In particular, miRNA-1246 and miRNA-150-3p were the most enriched strands by more than 150-folds in the 5-week storage period. Conclusions As storage progressed, numerous exosomal miRNAs accumulated in the RBC suspensions, which are informatically connected to multiple immuno-signaling pathways. MiRNA-1246 and miRNA-150-3p may be essential mediators impacting the immunoregulation functions of exosomes in RBC suspensions, considering their significant accumulating scales. Further research should therefore focus on the relationship between these miRNAs and TRIM.

Aquaporin 3 promotes human extravillous trophoblast migration and invasion

Problem Does aquaporin 3 (AQP3) affect the migration and invasion of human extravillous trophoblast (HTR8/Svneo) cells? Method of study A lentivirus infection system was used to construct stable cell lines with either AQP3 knockdown or overexpression. RT-PCR and western blotting were used to verify the efficiencies of AQP3 knockdown or overexpression in HTR8/Svneo cells at mRNA and protein levels, respectively. Cell Counting Kit-8 and flow cytometry assays were used to detect the influence of AQP3 knockdown or overexpression on proliferation and apoptosis of HTR8/Svneo cells. In addition, wound healing and Transwell invasion assays were used to detect the effects of AQP3 knockdown or overexpression on migration and invasion capabilities of HTR8/Svneo cells. An Agilent gene chip was used to screen for significant differentially expressed genes after AQP3 knockdown. Finally, mechanisms by which AQP3 influences the migration and invasion of HTR8/Svneo cells were explored using bioinformatic analysis. Results Compared with controls, migration and invasion capabilities of HTR8/Svneo cells were significantly reduced after AQP3 knockdown, and significantly increased after AQP3 overexpression. Subsequent bioinformatic analysis of gene chip expression profiles indicated downregulation of genes related to adhesion such as PDGF-B, as well as signaling pathways (such as PIK3/AKT, NF-κB, and TNF) after AQP3 knockdown. Conclusions AQP3 could significantly promote migration and invasion capabilities of human extravillous trophoblasts, it may mediate embryo invasion and adhesion to endometrium by regulating PDGF-B, PIK3/AKT signaling pathways, although this requires further verification.

Aquaporin 3 promotes human extravillous trophoblast migration and invasion

Problem: Does aquaporin 3 (AQP3) affect the migration and invasion of human extravillous trophoblast (HTR8/Svneo) cells?Method of Study: A lentivirus infection system was used to construct stable cell lines with either AQP3 knockdown or overexpression. RT-PCR and western blotting were used to verify the efficiencies of AQP3 knockdown or overexpression in HTR8/Svneo cells at mRNA and protein levels, respectively. Cell Counting Kit-8 and flow cytometry assays were used to detect the influence of AQP3 knockdown or overexpression on proliferation and apoptosis of HTR8/Svneo cells. In addition, wound healing and Transwell invasion assays were used to detect the effects of AQP3 knockdown or overexpression on migration and invasion capabilities of HTR8/Svneo cells. An Agilent gene chip was used to screen for significant differentially expressed genes after AQP3 knockdown. Finally, mechanisms by which AQP3 influences the migration and invasion of HTR8/Svneo cells were explored using bioinformatic analysis.Results: Compared with controls, migration and invasion capabilities of HTR8/Svneo cells were significantly reduced after AQP3 knockdown, and significantly increased after AQP3 overexpression. Subsequent bioinformatic analysis of gene chip expression profiles indicated downregulation of genes related to adhesion such as PDGF-B, as well as signaling pathways (such as PIK3/AKT, NF-κB, and TNF) after AQP3 knockdown. Conclusions: AQP3 could significantly promote migration and invasion capabilities of human extravillous trophoblasts, it may mediate embryo invasion and adhesion to endometrium by regulating PDGF-B, PIK3/AKT signaling pathways, although this requires further verification.