Intracellular Localization and Sustained Prodrug Cell Killing Activity of TAT-HSVTK Fusion Protein in Hepatocelullar Carcinoma Cells

Limin Cao,Jin Si,Weiyu Wang,Xiaorong Zhao,Xiaomei Yuan,Huifen Zhu,Xiaolong Wu,Jianzhong Zhu,Guanxin Shen 한국분자세포생물학회 2006 Molecules and cells Vol.21 No.1

Gene therapy with nonviral vectors using the suicide gene/prodrug activating system of herpes simplex virus type-1 thymidine kinase (HSV1-TK)/ganciclovir (GCV) is inefficient in killing malignant tumor cells due to two major factors: (a) an unsatisfactory bystander effect; (b) short-lived expression of the protein. To study the capacity of the protein transduction domain (PTD) of HIV-1 TAT protein to enhance HSV1-TK/GCV cancer gene therapy, we constructed three fusion proteins TAT-TK, TK-TAT and TK. TATTK retained as much enzyme activity as TK, whereas that of TK-TAT was much lower. TAT-TK can enter HepG2 cells and much of it is translocated to the nucleus. The transduced HepG2 cells are killed by exogenously added GCV and have bystander effects on untransduced HepG2 cells. Most importantly, the introduced recombinant protein is stable and remains functional for several days at least, probably because nuclear localization protects it from the cytoplasmic degradation machinery and provides access to the nuclear transcription machinery. Our results indicate that TAT fusion proteins traffic intercellularly and have enhanced stability and prodrug cell killing activity. We conclude that TAT has potential for enhancing enzyme prodrug treatment of liver cancers.

Comparative transcriptome analysis of wing discs from Bombyx mori and Bombyx mandarina

Feng Yongjie,Kumar Dhiraj,Hu Xiaolong,Zhang Yiling,Zhu Min,Xue Renyu,Cao Guangli,Gong Chengliang 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.2

The insect wing is developed from the wing imaginal disc which is designed from the embryonic ectoderm. To get insight into gene expression profiles in wing discs of Bombyx mori during metamorphosis, we compared the gene expression in the wing between B. mori and B. mandarina moth through RNA-seq. Out of total valid reads identified from the 5th day of 5th instar larvae of silkworm (L5), 7th day of pupae (P7), 1st day of moth (M1) and 1st day of wild silkworm moth (WM1), 20,092,004, 29,251,647, 24,654,695 and 19,753,089 reads were mapped to the mRNA reference sequences of silkworm, respectively. 9229, 7048, 9268 and 6701 differentially expressed genes (DEGs) were respectively recorded in P7 vs L5, M1 vs P7, M1 vs L5 and WM1 vs M1. Further, the peroxisome, ribosome, endocytosis and oxidative phosphorylation pathways were significantly regulated in the metamorphosis of the silkworm. Our study identified 16 orthologous genes with a positive selection from M1, which might be subjected to artificial selection in the domestication of B. mori and would play vital roles in the flight of B. mandarina.

Aspirin-Triggered Resolvin D1 Inhibits TGF-β1-Induced EndMT through Increasing the Expression of Smad7 and Is Closely Related to Oxidative Stress

Shu, Yusheng,Liu, Yu,Li, Xinxin,Cao, Ling,Yuan, Xiaolong,Li, Wenhui,Cao, Qianqian The Korean Society of Applied Pharmacology 2016 Biomolecules & Therapeutics(구 응용약물학회지) Vol.24 No.2

The endothelial-mesenchymal transition (EndMT) is known to be involved in the transformation of vascular endothelial cells to mesenchymal cells. EndMT has been confirmed that occur in various pathologic conditions. Transforming growth factor ${\beta}1$ (TGF-${\beta}1$) is a potent stimulator of the vascular endothelial to mesenchymal transition (EMT). Aspirin-triggered resolvin D1 (AT-RvD1) has been known to be involved in the resolution of inflammation, but whether it has effects on TGF-${\beta}1$-induced EndMT is not yet clear. Therefore, we investigated the effects of AT-RvD1 on the EndMT of human umbilical vein vascular endothelial cells line (HUVECs). Treatment with TGF-${\beta}1$ reduced the expression of Nrf2 and enhanced the level of F-actin, which is associated with paracellular permeability. The expression of endothelial marker VE-cadherin in HUVEC cells was reduced, and the expression of mesenchymal marker vimentin was enhanced. AT-RvD1 restored the expression of Nrf2 and vimentin and enhanced the expression of VE-cadherin. AT-RvD1 did also affect the migration of HUVEC cells. Inhibitory ${\kappa}B$ kinase 16 (IKK 16), which is known to inhibit the NF-${\kappa}B$ pathway, had an ability to increase the expression of Nrf2 and was associated with the inhibition effect of AT-RvD1 on TGF-${\beta}1$-induced EndMT, but it had no effect on TGF-${\beta}1$-induced EndMT alone. Smad7, which is a key regulator of TGF-${\beta}$/Smads signaling by negative feedback loops, was significantly increased with the treatment of AT-RvD1. These results suggest the possibility that AT-RvD1 suppresses the TGF-${\beta}1$-induced EndMT through increasing the expression of Smad7 and is closely related to oxidative stress.

Study of gut bacterial diversity of Bombyx mandarina and Bombyx mori through 16S rRNA gene sequencing

Dhiraj Kumar,Zhenli Sun,Guangli Cao,Renyu Xue,Xiaolong Hu,Chengliang Gong 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.2

The wild silkworm B. mandarina is living in the natural environment has a strong stress resistance and adaptability after harsh natural selection. The indoor rearing or domestication of the wild silkworm under artificial custody for long period deteriorates stress resistance and ecological adaptability. Therefore, we aimed to investigate the effects of artificial domestication and evolutionary pressure on the gut bacterial diversity of B. mandarina and B. mori. The intestinal content of 6th day of fifth instar B. mandarina and B. mori larvae were analyzed by sequencing of the 16S rRNA gene through Illumina miseq sequencing technology. The outcome of the study revealed that abundance of predominant bacteria of phylum Firmicutes were respectively 81.40% and 81.85% in the late fifth instar silkworm larvae (6th day) of B. mandarina and B. mori. In Firmicutes, abundance of predominant bacterial genus Enterococcus in B. mandarina (69.73%) was comparatively higher than B. mori (48.99%). The genus Advenella belongs to phylum Proteobacteria was recorded only in B. mandarina (11.54%). The abundance of Unclassified_Peptostreptococcaceae, Methanobrevibacter, Ignatzschineria, Petrimonas and Proteiniphilum in B. mandarina were between 0.12 and 0.17%, nevertheless, these bacterial genera were not detected in B. mori. The abundance of genera Lactococcus, Bacillus and Pseudomonas in B. mori (17.73%, 5.02%, and 1.61%) were remarkably higher than B. mandarina (0.15%, 0.54% and 0.45%). These results indicated that substantial difference was observed between the intestinal bacteria of B. mori and B. mandarina population, and structure of the intestinal bacteria could be affected by the artificial domestication and evolutionary pressure.

Effect of Particle Size and Dispersion on Dielectric Properties in ZnO/Epoxy Resin Composites

Wenhu Yang,Ran Yi,Xu Yang,Man Xu,Sisi Hui,Xiaolong Cao 한국전기전자재료학회 2012 Transactions on Electrical and Electronic Material Vol.13 No.3

In this paper, ZnO-Epoxy nanocomposites (NEP) were prepared and epoxy composites that contain 5 wt% micro ZnO (MEP) and deliberately not well dispersed nano ZnO (NDNEP) were also prepared for purpose of comparison. The effects of the particle size and dispersion of ZnO on dielectric properties of epoxy resin were chiefly studied. Test results showed that: at a loading of 5 wt%, the three epoxy composites seem to have no significant difference on resistivity compared to epoxy resin; Dielectric constants of all the epoxy composites are also basically the same but they are bigger compared to that of the pure epoxy resin (unfilled); Dielectric dissipation factors (tanδ) of NDNEP is greater than that of NEP and MEP. NEP has the minimum dielectric loss factor, whereas dielectric loss factors of the three epoxy composites are larger than that of the pure epoxy resin. The decreasing order of electrical breakdown strength for the three epoxy composites and for the pure epoxy resin is as follows: NEP>MEP>NDNEP>EP. Finally,in order to explain the experimental results the aggregation interface phase was proposed. Furthermore, addition of well dispersed nano filler has proved to have a positive effect on the improvement of the dielectric properties of epoxy resin.

Coal dust exposure induces proliferation and migration of human bronchial epithelial cells

Li Amin,Zhang Yinci,Wang Ruikai,Xu Ruyue,Ma Yongfang,Song Li,Cao Weiya,Xiaolong Tang 대한독성 유전단백체 학회 2023 Molecular & cellular toxicology Vol.19 No.2

Background Coal dust exposure has caused a variety of lung diseases. In addition to genotoxicity and cytotoxicity, other biological changes caused by coal dust (CD) exposure need further study. Objective To observe the cellular transformation eff ects of CD exposure and explore its underlying molecular mechanism, human bronchial epithelial cells (BEAS-2B) were cultured with continuous CD exposure. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, colony formation assay, wound healing assay, next-generation sequencing (NGS) and western blotting were performed to observe the cell proliferation, migration, genomic transcription and pathological signaling pathways. Results We demonstrated that BEAS-2B cells with long-term chronic CD exposure show accelerated proliferation rate and enhanced migration ability, and have altered gene expression profi les and aberrant activation of EGFR/Raf/ERK and PI3K/ AKT/mTOR pathways. Conclusions The results indicate that chronic CD exposure could induce abnormal proliferation and migration of BEAS-2B cells, lead to the transformation potential of human bronchial epithelial cells.

Effect of Particle Size and Dispersion on Dielectric Properties in ZnO/Epoxy Resin Composites

Yang, Wenhu,Yi, Ran,Yang, Xu,Xu, Man,Hui, Sisi,Cao, Xiaolong The Korean Institute of Electrical and Electronic 2012 Transactions on Electrical and Electronic Material Vol.13 No.3

In this paper, ZnO-Epoxy nanocomposites (NEP) were prepared and epoxy composites that contain 5 wt% micro ZnO (MEP) and deliberately not well dispersed nano ZnO (NDNEP) were also prepared for purpose of comparison. The effects of the particle size and dispersion of ZnO on dielectric properties of epoxy resin were chiefly studied. Test results showed that: at a loading of 5 wt%, the three epoxy composites seem to have no significant difference on resistivity compared to epoxy resin; Dielectric constants of all the epoxy composites are also basically the same but they are bigger compared to that of the pure epoxy resin (unfilled); Dielectric dissipation factors ($tan{\delta}$) of NDNEP is greater than that of NEP and MEP. NEP has the minimum dielectric loss factor, whereas dielectric loss factors of the three epoxy composites are larger than that of the pure epoxy resin. The decreasing order of electrical breakdown strength for the three epoxy composites and for the pure epoxy resin is as follows: NEP>MEP>NDNEP>EP. Finally, in order to explain the experimental results the aggregation interface phase was proposed. Furthermore, addition of well dispersed nano filler has proved to have a positive effect on the improvement of the dielectric properties of epoxy resin.

Cell proliferation can be modulated by receptor tyrosine kinase-like orphan receptor 2 in the silkworm, Bombyx mori

Yongjie Feng,Wei Liu,Dhiraj Kumar,Min Zhu,Renyu Xue,Guangli Cao,Xiaolong Hu,Chengliang Gong 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.4

The receptor tyrosine kinase-like orphan receptor 2 (Ror2) is involved in the Wnt/β-catenin signaling pathway which regulates cell proliferation, polarity, differentiation, migration, metabolism and survival. However, the function of Ror2 in the silkworm Bombyx mori is still undisclosed. In the present investigation, we have made an effort to clone the silkworm Ror2 gene (BmRor2). The sequence analysis showed that the open reading frame (ORF) was 1914 bp in size and encoded a protein with the conserved domains of Ror2 protein. The qRT-PCR results indicated that the BmRor2 gene expression level was the highest in the head among all identified tis sues on 3rd day of the fifth instar larvae. In the gonads of the different development stages, the BmRor2 gene expression level was highest on the 4th day of the fourth instar larvae. The immunofluorescence assay indicated that the BmRor2 protein was located at the cytomembrane. The effects of BmRor2 protein on the expression levels of genes related to TGF-β, Hippo, JAK-STAT and Notch signaling pathways were investigated by qRT-PCR. The expression levels of crumbs (crb), warts (wts), α-catenin (cat), four-jointed (fj), decapentaplegic (dpp), kibra ortholog (kibra), serrate (serr) and c-myc (myc) genes were down-regulated, whereas, suppressor of cytokine signaling 2 (socs 2) gene expression was up-regulated in the cultured BmN cells after the BmRor2 expression level was up-regulated. Further, cell proliferation was demoted and the size of cells was decreased when BmRor2 expression level was elevated. Our current finding recommended that BmRo2 can regulate TGF- β, Hippo, JAKSTAT, and Notch signaling pathways, and affect cell proliferation and size.

Transient propagation of BmLV and dysregulation of gene expression in nontarget cells following BmLV infection

Feng Yongjie,Zhang Xing,Kumar Dhiraj,Kuang Sulan,Liu Bo,Hu Xiaolong,Zhu Min,Liang Zi,Cao Guangli,Xue Renyu,Gong Chengliang 한국응용곤충학회 2021 Journal of Asia-Pacific Entomology Vol.24 No.3

Bombyx mori latent virus (BmLV), a novel positive-strand RNA virus was first identified in the B. mori cultured BmN cell line. Whether the infectivity of BmLV to silkworm larvae and non-silkworm cells is connected with dysregulation of gene expression are not well understood. A complete sequence of BmLV genomic RNA was identified and revealed that a fragment with 495 nt in length was deleted from the RNA-dependent RNA poly merase (RdRp) gene in some BmLV genomic RNAs. Studies on the infectivity of BmLV to nontarget cells showed that BmLV can infect silkworm larvae, Spodoptera frugiperda Sf-9 and H1299 lung cancer cells with transient propagation. The dysregulation of gene expression of Sf-9 cells followed by BmLV infection was analyzed. Out of 743 differentially expressed genes (DEGs), 300 were upregulated and 443 were downregulated. Gene Ontology (GO) analysis indicated the DEGs were enriched into oxidoreductase activity for CH-NH 2 group donors, gluta mate biosynthetic process, response to stress and proteasome core complex. KEGG enrichment analysis showed that DEGs were mainly enriched into sulfur metabolism, RNA degradation, proteasome, pentose and glucuronate interconversions. Undesirable nutrients and temperature factors contributed to the propagation of BmLV in Sf-9 cells. Additionally, the Imd and RNAi pathways were activated by BmLV infection without stimulating Toll and JAK-STAT pathways. Therefore, it is suggested that BmLV is originated from plants, which can enter nontarget cells with transient propagation. The transient infection of BmLV may not only be regulated by Imd and RNAi immune pathways but also mediated by dysregulation of gene expression.