LncRNA MALAT1 facilitates Parkinson's disease progression by increasing SOCS3 promoter methylation.

BACKGROUND: Long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) has been shown to be involved in Parkinson's disease (PD) progression, but its mechanism needs to be further explored. METHODS: Mice were injected with 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to induce PD mice models, and BV2 cells were treated with lipopolysaccharides (LPS) to mimic PD cell models. MALAT1 expression and suppressor of cytokine signaling 3 (SOCS3) protein level were examined using quantitative real-time PCR and western blot, respectively. Cell functions were tested by cell counting kit 8 assay and flow cytometry. The interaction between MALAT1 and SOCS3 was confirmed using RNA pull-down and RIP assays. RESULTS: MALAT1 was upregulated in MPTP-induced PD mice and LPS-induced BV2 cells. Silencing of MALAT1 increased viability, while inhibited apoptosis and inflammation in LPS-induced BV2 cells. Besides, MALAT1 enhanced the SOCS3 promoter methylation to decrease its expression by recruiting DNMT1, DNMT3A and DNMT3B. Furthermore, SOCS3 knockdown eliminated sh-MALAT1-mediated the inhibition effect on LPS-induced BV2 cell injury. In vivo, MALAT1 silencing ameliorated neurological impairment and neuroinflammation in MPTP-induced PD mice. CONCLUSION: Our data revealed that MALAT1 worsened PD processes via inhibiting SOCS3 expression by increasing its promoter methylation.

LncRNA SOX21-AS1 Promotes Activation of BV2 Cells via Epigenetical Silencing of SOCS3 and Aggravates Parkinson's Disease.

BACKGROUND: LncRNAs perform a crucial impact on microglia's activation in Parkinson's disease (PD). Here, our purpose was to probe the function and involved mechanism of lncRNA SOX21-AS1 on microglial activation in PD. METHODS: Mice were treated with MPTP, and BV2 cells were treated with LPS/ATP to build PD animal and cell models. Genes' expression was measured using RT-qPCR, immunoblotting, and IHC stain. ELISA was applied for testing inflammatory factors' levels. Cell viability and apoptosis were tested using kits. RIP and RNA pull-down assay were utilized for monitoring the bond of SOX21-AS1 to EZH2, and ChIP was applied for affirming the bond between EZH2 and SOCS3's promoter. RESULTS: The expression of SOX21-AS1 and SOCS3 was abnormal in PD cell and animal models. Inhibition of SOX21-AS1 repressed LPS/ATP-induced activation in BV2 cells and nerve damage caused by activated BV2 cells, alleviating the pathological features of PD mice. Further studies found that SOX21-AS1 epigenetically inhibited SOCS3 by recruiting EZH2 to SOCS3 promoter. SOX21-AS1 overexpression partially offset the repressive impact of SOCS3 enhancement on BV2 cell activation and the protective effect on nerve cells. CONCLUSION: SOX21-AS1 enhances LPS/ATP-induced activation of BV2 cells and nerve damage caused by activated BV2 cells though recruiting EZH2 to SOCS3's promoter, thereby alleviating PD progression. Our research supplies new potential target for curing PD.

KIF11 As a Potential Pan-Cancer Immunological Biomarker Encompassing the Disease Staging, Prognoses, Tumor Microenvironment, and Therapeutic Responses.

KIF11 is one of the 45 family members of kinesin superfamily proteins that functions as a motor protein in mitosis. Emerging evidence revealed that KIF11 plays pivotal roles in cancer initiation, development, and progression. However, the prognostic, oncological, and immunological values of KIF11 have not been comprehensively explored in pan-cancer. In present study, we comprehensively interrogated the role of KIF11 in tumor progression, tumor stemness, genomic heterogeneity, tumor immune infiltration, immune evasion, therapy response, and prognosis of cohorts from various cancer types. In general, KIF11 was significantly upregulated in tumors compared with paired normal tissues. KIF11 showed strong relationships with pathological stage, prognosis, tumor stemness, genomic heterogeneity, neoantigens, ESTIMATE, immune checkpoint, and drug sensitivity. The methylation level of KIF11 decreased in most cancers and was correlated with the survival probability in different human cancers. The expression of KIF11 was diverse in different molecular and immune subtypes and remarkably correlated with immune cell infiltration in the tumor microenvironment. Comparative study revealed that KIF11 was a powerful biomarker and associated with immune, targeted, and chemotherapeutic outcomes in various cancers. In addition, KIF11 interaction and coexpression networks mainly participated in the regulation of cell cycle, cell division, p53 signaling pathway, DNA repair and recombination, chromatin organization, antigen processing and presentation, and drug resistance. Our pan-cancer analysis provides a comprehensive understanding of the functions of KIF11 in oncogenesis, progression, and therapy in different cancers. KIF11 may serve as a potential prognostic and immunological pan-cancer biomarker. Moreover, KIF11 could be a novel target for tumor immunotherapy.

A MYB/bHLH complex regulates tissue-specific anthocyanin biosynthesis in the inner pericarp of red-centered kiwifruit Actinidia chinensis cv. Hongyang.

Many Actinidia cultivars are characterized by anthocyanin accumulation, specifically in the inner pericarp, but the underlying regulatory mechanism remains elusive. Here we report two interacting transcription factors, AcMYB123 and AcbHLH42, that regulate tissue-specific anthocyanin biosynthesis in the inner pericarp of Actinidia chinensis cv. Hongyang. Through transcriptome profiling analysis we identified five MYB and three bHLH transcription factors that were upregulated in the inner pericarp. We show that the combinatorial action of two of them, AcMYB123 and AcbHLH42, is required for activating promoters of AcANS and AcF3GT1 that encode the dedicated enzymes for anthocyanin biosynthesis. The presence of anthocyanin in the inner pericarp appears to be tightly associated with elevated expression of AcMYB123 and AcbHLH42. RNA interference repression of AcMYB123, AcbHLH42, AcF3GT1 and AcANS in 'Hongyang' fruits resulted in significantly reduced anthocyanin biosynthesis. Using both transient assays in Nicotiana tabacum leaves or Actinidia arguta fruits and stable transformation in Arabidopsis, we demonstrate that co-expression of AcMYB123 and AcbHLH42 is a prerequisite for anthocyanin production by activating transcription of AcF3GT1 and AcANS or the homologous genes. Phylogenetic analysis suggests that AcMYB123 or AcbHLH42 are closely related to TT2 or TT8, respectively, which determines proanthocyanidin biosynthesis in Arabidopsis, and to anthocyanin regulators in monocots rather than regulators in dicots. All these experimental results suggest that AcMYB123 and AcbHLH42 are the components involved in spatiotemporal regulation of anthocyanin biosynthesis specifically in the inner pericarp of kiwifruit.

Comprehensive Transcriptome Profiling Reveals Long Noncoding RNA Expression and Alternative Splicing Regulation during Fruit Development and Ripening in Kiwifruit (Actinidia chinensis).

Genomic and transcriptomic data on kiwifruit (Actinidia chinensis) in public databases are very limited despite its nutritional and economic value. Previously, we have constructed and sequenced nine fruit RNA-Seq libraries of A. chinensis "Hongyang" at immature, mature, and postharvest ripening stages of fruit development, and generated over 66.2 million paired-end and 24.4 million single-end reads. From this dataset, here we have identified 7051 long noncoding RNAs (lncRNAs), 29,327 alternative splicing (AS) events and 2980 novel protein-coding genes that were not annotated in the draft genome of "Hongyang." AS events were demonstrated in genes involved in the synthesis of nutritional metabolites in fruit, such as ascorbic acids, carotenoids, anthocyanins, and chlorophylls, and also in genes in the ethylene signaling pathway, which plays an indispensable role in fruit ripening. Additionally, transcriptome profiles and the contents of sugars, organic and main amino acids were compared between immature, mature, and postharvest ripening stages in kiwifruits. A total of 5931 differentially expressed genes were identified, including those associated with the metabolism of sugar, organic acid, and main amino acids. The data generated in this study provide a foundation for further studies of fruit development and ripening in kiwifruit, and identify candidate genes and regulatory elements that could serve as targets for improving important agronomic traits through marker assisted breeding and biotechnology.

Comparative studies on zirconia and graphene composites obtained by one-step and stepwise electrodeposition for deoxyribonucleic acid sensing.

In this paper, the comparison of two kinds of electrochemically reduced graphene oxide (ERGNO) and zirconia composites, obtained by one-step (ZrO2-ERGNO) and stepwise (ZrO2/ERGNO) electrodeposition for DNA sensing, is systematically studied. The resulting composites were characterized by scanning electron microscopy, cyclic voltammetry, and differential pulse voltammetry. The results indicated that the ZrO2-ERGNO presented fine globular nanostructure. However, ZrO2/ERGNO presented agglomerate massive microstructure due to the absence of the oxygen-containing groups of graphene oxide, confirming the oxygen-containing groups provided a better affinity for the deposition of ZrO2. Due to the strong binding of the phosphate groups of DNA with the zirconia film, DNA probes were attached on the ZrO2-based composites. ZrO2-ERGNO/Au owning fine nanostructure presented larger surface area than microstructured ZrO2/ERGNO/Au. Moreover, compared with microstructured ZrO2/ERGNO, the nanostructured ZrO2-ERGNO provided more accessible space for immobilized DNA probe hybridization with target sequence, which consequently resulted in higher hybridization efficiency. Therefore, the ZrO2-ERGNO was chosen for fabricating DNA sensor with a limit of detection 1.21×10(-14) mol L(-1).

Electrochemical synthesis of Fe2O3 on graphene matrix for indicator-free impedimetric aptasensing.

Herein, an electrochemical platform was employed for the detection of protein. Fe2O3 was electrochemically deposited on graphene modified glassy carbon electrode surface. Electrodeposition conditions, such as temperature, and time, were optimized for controlling morphologies and electrochemical activities of Fe2O3. Negatively charged lysozyme-binding aptamer (LBA) was immobilized on positively charged Fe2O3 (isoelectric point ≈ 7.0) via electrostatic interaction. Electrochemical impedance spectroscopy was adopted for indicator-free detection of lysozyme. The LBA on the outermost layer would catch lysozyme in solution by physical affinity, which induced the increase of impedimetric signals. In this strategy, a wide detection range (0.5 ng mL(-1)-5 µg mL(-1)) and low detection limit (0.16 ng mL(-1)) for model target lysozyme was obtained. The results showed that indicator-free impedimetric aptasensing strategy had good sensitivity and selectivity.

Electrochemical impedimetric DNA sensing based on multi-walled carbon nanotubes-SnO2-chitosan nanocomposite.

A sensitive electrochemical impedimetric DNA biosensor based on the integration of tin oxide (SnO2) nanoparticles, chitosan (CHIT) and multi-walled carbon nanotubes (MWNTs) is presented in this paper. The MWNTs-SnO2-CHIT composite modified gold electrode was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Compared with individual MWNTs-CHIT, SnO2-CHIT and bare gold electrode, this composite showed the most obvious electrochemical signal of the redox probe [Fe(CN)6](3-/4-). According to the change of the electron transfer resistance (R(et)) induced by the hybridization, target DNA was successfully detected via EIS. This DNA electrochemical biosensor was applied to detect phosphinothricin acetyltransferase (PAT) gene in transgenic corn. The synergistic effect of the MWNTs-SnO2-CHIT remarkably enhanced DNA immobilization and hybridization detection. The dynamic detection range was from 1.0×10(-11) mol/L to 1.0×10(-6) mol/L with a detection limit of 2.5×10(-12) mol/L. This sensing platform showed inner advantage, such as simplicity, good stability, and high sensitivity.

Direct and freely switchable detection of target genes engineered by reduced graphene oxide-poly(m-aminobenzenesulfonic acid) nanocomposite via synchronous pulse electrosynthesis.

A novel one-step electrochemical synthesis of the reduced graphene oxide and poly(m-aminobenzenesulfonic acid, ABSA) nanocomposite (PABSA-rGNO) via pulse potentiostatic method (PPM) for direct and freely switchable detection of target genes is presented. Unlike most electrochemical preparation of hybrids based on rGNO and polymer, electrochemical synthesis of PABSA (during the pulse electropolymerization period of PPM) and electrochemical reduction of rGNO (during the resting period of PPM), in this paper, were alternately performed. The total progress synchronously resulted in PABSA-rGNO nanocomposite. This nanocomposite was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), Fourier Transform infrared spectroscopy (FT-IR), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The PABSA-rGNO nanocomposite integrated graphene (a single-atom thick, two-dimensional sheet of sp(2) bonded conjugated carbon) with PABSA (owning rich-conjugated structures, functional groups, and excellent electrochemical activity), which could serve as an ideal electrode material for biosensing and electrochemical cell, etc. As an example, the immobilization of the specific probe DNA was successfully conducted via the noncovalent method due to the π-π* interaction between conjugated nanocomposite and DNA bases. The hybridization between the probe DNA and target DNA induced the product dsDNA to be released from conjugated nanocomposite, accompanied with the self-signal regeneration of nanocomposite ("signal-on"). The self-signal changes served as a powerful tool for direct and freely switchable detection of different target genes, and the synergistic effect of PABSA-rGNO nanocomposite effectively improved the sensitivity for the target DNA detection.