LncRNA TUG1 Promotes Hepatocellular Carcinoma Migration and Invasion Via Targeting the miR-137/AKT2 Axis.

Background: The current study aimed to investigate the effects of TUG1 on the migration and invasion of hepatoma cells. Materials and Methods: The expressions of TUG1, miR-137, and AKT2 were detected in hepatoma tissues and cells by performing quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The correlations among TUG1, miR-137, and AKT2 were predicted by bioinformatics analysis and confirmed by dual-luciferase reporter assay, and Pearson test was performed to analyze their relevance. The effects of TUG1, miR-137, and AKT2 on viability, migration, and invasion of transfected hepatoma cells were detected by CCK-8, wound scratch, and Transwell. Epithelial-mesenchymal transition (EMT)-related protein levels were determined by Western blot and qRT-PCR. Results: TUG1 was highly expressed in hepatoma tissues and cells. Silencing TUG1 expression inhibited the viability, migration, and invasion of hepatoma cells. TUG1 targeted miR-137 and the two was negatively correlated, and silencing TUG1 expression inhibited the effects of low-expressed miR-137 on promoting proliferation, migration, and invasion of hepatoma cells. AKT2 was predicted to be the target gene for miR-137, and the two were negatively correlated. Moreover, inhibiting miR-137 expression promoted the expression of MMP2, MMP9, and N-cadherin and inhibited E-cadherin expression, while silencing TUG1 expression reversed the effects of low-expressed miR-137 on EMT-related protein levels. Conclusion: LncRNA TUG1 promotes hepatocellular carcinoma migration and invasion through targeting the miR-137/AKT2 axis.

Complete mitochondrial genome of the common cutworm Spodoptera litura (Lepidoptera: Noctuidae).

We determined the complete mitochondrial genome of the common cutworm Spodoptera litura (Lepidoptera: Noctuidae), which is one of the most destructive polyphagous insect pests worldwide. The genome is 15,383 bp in length (GenBank accession number: KF701043) with an A+T content of 81.08%, and contains 37 typical animal mitochondrial genes (13 protein-coding genes, 2 rRNA genes and 22 tRNA genes) with the typical arrangement found in Lepidoptera. All the protein-coding genes (PCGs) start with ATN start codon except for cox1, which begins with CGA. Eight PCGs stop with complete termination codons (TAA or TAG), whereas five PCGs use incomplete stop codon T. The A+T-rich region is located between rrnS and trnM with a length of 326 bp and an A+T content of 93.87%, and harbors three tandem repeat elements.

Complete mitochondrial genome of the oriental armyworm Mythimna separata (Walker) (Lepidoptera: Noctuidae).

We determined the complete mitochondrial genome of the oriental armyworm Mythimna separata (Walker) (Lepidoptera: Noctuidae), which is one of the serious cereal pests in Asia and Australia. The circular genome of 15,332 bp in length contains 37 typical animal mitochondrial genes and a non-coding A+T-rich region. Its gene content and order are typical of lepidopteran mitochondrial genomes described to date. All protein-coding genes (PCGs) start with an ATN codon except for cox1 and nad1, which use CGA and TTG as their start codon, respectively. Ten PCGs use complete stop codon TAA, whereas three PCGs end with single T. The A+T-region is located between rrnS and trnM with a length of 374 bp. The mitochondrial genome sequence benefits future studies of molecular phylogenetics and pest control.

Polysaccharide from Phellinus linteus induces S-phase arrest in HepG2 cells by decreasing calreticulin expression and activating the P27kip1-cyclin A/D1/E-CDK2 pathway.

ETHNOPHARMACOLOGY RELEVANCE: Our previous study showed that the proteoglycan P1 from Phellinus linteus (Mesima) exhibits significant anti-tumor activity against human hepatocellular carcinoma cells (HepG2); however, its molecular mechanism remains unknown. This study aims to provide insights into the mechanism of the anti-tumor activity of P1 against HepG2 cells. METHODS: We examined the effects of P1 on HepG2 cell proliferation in vitro and in vivo. Flow cytometry was used to analyze the cell cycle distribution and apoptosis. Proteomic analysis, real-time (RT)-PCR, and Western blot were carried out to observe the expression of several cell cycle control proteins in HepG2 cells. RESULTS: Both the volume and the weight of solid tumors were significantly decreased in P1-treated mice (200mg/kg) compared with the control. The HepG2 cells in the P1-treated tumors were significantly decreased, irregularly shaped, and smaller. P1 slightly increased the body weight of the tumor-bearing mice, which indicates that P1 is nontoxic to mammals at 200mg/kg. P1 also caused a significant dose-dependent increase in S phase arrest, but no apoptosis was observed in HepG2 cells. The results of the proteomic analysis, RT-PCR, and Western blot analysis showed that significantly downregulated expression of calreticulin, cyclin D1, cyclin E, and CDK2 and upregulated expression of P27 kip1 and cyclin A in the P1-treated HepG2 cells (200 µg/ml). CONCLUSION: These results suggest that calreticulin expression and the P27 kip1-cyclin A/D1/E-CDK2 pathway were involved in P1-induced S-phase cell cycle arrest in HepG2 cells.

Proteome analysis of silkworm, Bombyx mori, larval gonads: characterization of proteins involved in sexual dimorphism and gametogenesis.

Sexual dimorphism is initialed by the components of the sex determination pathway and is most evident in gonads and germ cells. Although striking dimorphic expressions have been detected at the transcriptional level between the silkworm larval testis and the ovary, the sex-dimorphic expressions at the protein level have not yet been well characterized. The proteome of silkworm larval gonads was investigated using a shotgun-based identification. A total of 286 and 205 nonredundant proteins were identified from the silkworm testis and ovary, respectively, with a false discovery rate (FDR) lower than 1%. Only 40 and 16 proteins were previously identified, and 246 and 189 proteins were newly identified in the silkworm testis and the ovary, respectively. The gametogenesis mechanism of silkworm was demonstrated using the protein expression profile and bioinformatics analysis. Cellular retinoic acid binding protein (CRABP) showed to be highly abundant in testis, while tubulins were abundant in ovary. Several homologies of Drosophila essential proteins for gametogenesis were identified in silkworm, such as male meiotic arrest gene product ALY and VISMAY in testis, and maternal mRNA localization protein exuperantia and SQUID in ovary. The gene ontology (GO) annotation and pathway analysis provide system-level insights into the sexual dimorphism and gametogenesis.

Shotgun proteomic analysis on the embryos of silkworm Bombyx mori at the end of organogenesis.

Embryonic development of silkworm, Bombyx mori is a process of systematical expression of genes and proteins which is dominated by complex regulatory networks. To gain comprehensive insight into the molecular basis of embryonic development and its regulation mechanisms, the proteome profile of the B. mori embryos at the end of organogenesis (tubercle appearance stage, TA) was characterized using LTQ-Orbitrap mass spectrometer. Totally 963 proteins were identified with a false discovery rate (FDR) of 0.12%. They were involved in embryonic development, chemoreception, and stimuli response and so forth. The proteins with the largest number of identified unique peptides, implying their possibly higher abundance, were involved in heat shock response, lipid transport and metabolism, and apoptosis. It was consistent with the physiological status of embryo at the end of organogenesis. Many functionally important proteins were identified for the first time in B. mori embryo such as the progesterone receptor membrane component 2, antennal binding protein, sericotropin, and molting fluid carboxypeptidase A (MF-CPA). 253 (26.27%) specific proteins in TA versus labrum appearance stage (LA, four days before TA) embryos were identified, which were mainly associated with musculature, nervous system, and chemoreception system. They disclosed the differential temporal and spatial expression of proteins in the process of organogenesis. The relative mRNA levels of fifteen identified proteins in the two experimented stages were also compared using quantitative reverse transcription PCR (qRT-PCR) and showed some inconsistencies with protein expression. Gene Ontology (GO) annotation of the identified proteins showed that the most proteome representations were in the categories of "binding" and "catalytic" in molecular function, and "cellular process" and "metabolic process" in biological process.

Proteomic and bioinformatic analysis on endocrine organs of domesticated silkworm, Bombyx mori L. for a comprehensive understanding of their roles and relations.

Three organs of silkworm larva endocrine system, including brain (Br), subesophageal ganglion (SG) and prothoracic glands (PG), were studied employing shotgun LC-MS/MS combined with bioinformatic analysis to comprehensively understand their roles and relations. Totally, 3430, 2683, and 3395 proteins were identified including 1885 common and 652, 253, and 790 organ-specific ones in Br, SG, and PG, respectively. Identified common-expressed proteins indicated the existence of intrinsic complex interactions among these parts of endocrine system. Most of the reputed organs-specific proteins were identified by this approach. KEGG pathway analysis showed 162 same pathways among the 169, 164, and 171 relating Br, SG, and PG. This analysis revealed functional similarities with exceptional resemblance in their metabolism and signaling pathways of the three organs. On the other hand, 70, 57, and 114 organ-specific enzymes related pathways were detected for Br, SG, and PG confirming their functional differences. These results reveal a cooperative mechanism among the three endocrine organs in regulating various physiological and developmental events, and also suggest that the organ-specific proteins might be the fundamental factors responsible for the functional differentiation of these organs.

Comparison of transformation efficiency of piggyBac transposon among three different silkworm Bombyx mori Strains.

The transformation rate of three different strains of silkworm Bombyx mori was compared after the introduction of enhanced green fluorescence protein (EGFP)-encoding genes into the silkworm eggs by microinjection of a mixture of piggyBac vector and helper plasmid containing a transposase-encoding sequence. Although there were no significant differences among the three strains in the percentages of fertile moths in microinjected eggs (P=0.1258), the percentages of G(0) transformed moths in fertile moths and injected eggs were both significantly different (P=0.01368 and P=0.02398, respectively). The transformation rate of the Nistari strain (Indian strain) was significantly higher than that of the other two strains, Golden-yellow-cocoon (Vietnamese strain) and Jiaqiu (Chinese strain), which had similar rate. These results indicate that the transformation efficiency of the piggyBac-based system might vary with silkworm strains with different genetic backgrounds. The presence of endogenous piggyBac-like elements might be an important factor influencing the transformation efficiency of introduced piggyBac-derived vectors, and the diverse amount and activation in different silkworm strains might account for the significant differences.