A novel granulocyte-specific α integrin is essential for cellular immunity in the silkworm Bombyx mori

Pan

et al. 2018

Cell Death Dis

Self Cite

Glioma is the most common and malignant form of primary brain tumour, and is characterised by high proliferation and extensive invasion and neurological destruction. Demethylzeylasteral (T-96), which is extracted from Tripterygium wilfordii, is considered to have immunosuppressive, anti-inflammatory and anti-angiogenic effects. Here, the anti-tumour effect of T-96 on glioma was evaluated. Our results demonstrated that T-96 significantly inhibited glioma cell growth and induced cell cycle arrest in G1 phase but did not induce apoptosis. Cell invasion and migration were dramatically suppressed after treatment with T-96. Almost all genes related to cell cycle and DNA replication were downregulated after treatment with T-96. Our results showed that miR-30e-5p was noticeably upregulated after T-96 treatment, and MYBL2, which is involved in cell cycle progression and is a target gene of miR-30e-5p, was significantly reduced in synchrony. Overexpression of MYBL2 partially rescued the T-96-induced inhibition of cell growth and proliferation. Moreover, a miR-30e-5p antagomir significantly reduced the upregulation of miR-30e-5p expression induced by T-96, leading to recovery of MYBL2 expression, and partially rescued the T-96-induced inhibition of cell growth and proliferation. More important, T-96 effectively upregulated miR-30e-5p expression and downregulated MYBL2 expression, thus inhibiting LN-229 cell tumour growth in a mouse model. These results indicated that T-96 might inhibit glioma cell growth by regulating the miR-30e-5p/MYBL2 axis. Our study demonstrated that T-96 might act as a promising agent for malignant glioma therapy.

Section: Methodsmentioning

confidence: 99%

Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

Pan

et al. 2018

Cell Death Dis

Self Cite

“…The primers used in this study are listed in Table 3. The recombinant plasmids (PET28a-Integrin b2 and PET28a-Integrin b3) were transformed into Escherichia coli Rosseta (DE3) cells and the expression of the recombinant proteins was induced by isopropyl b-D-1-thiogalactopyranoside as described previously (Zhang et al, 2014a). The recombinant proteins were purified using Ni-nitrilotriacetic acid (NTA) His Bind Resin (Novagen, Madison, WI, USA) according to the manufacturer's instructions.…”

Section: Antibody Preparationmentioning

confidence: 99%

“…Spherulocytes represent a small number of haemocytes and have an unknown function (Ribeiro & Brehélin, ). Based on the tissue microarray database (Xia et al ., ), several haemocyte‐specific candidate genes were identified (Park et al ., ; Tsubota et al ., ; Zhang et al ., , , b). In our previous studies, we found that three candidate promoters could drive reporter gene‐specific expression in haemocytes using the Autographa californica nuclear polyhedrosis virus (AcNPV) system (Zhang et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Transgenic characterization of two silkworm tissue‐specific promoters in the haemocyte plasmatocyte cells

Insect Molecular Biology

Weng

et al. 2017

Haemocytes play crucial roles in insect metabolism, metamorphosis, and innate immunity. As a model of lepidopteran insects, the silkworm is a useful model to study the functions of both haematopoiesis and haemocytes. Tissue-specific promoters are excellent tools for genetic manipulation and are widely used in fundamental biological research. Herein, two haemocyte-specific genes, Integrin β2 and Integrin β3, were confirmed. Promoter activities of Integrin β2 and Integrin β3 were evaluated by genetic manipulation. Quantitative real-time PCR and western blotting suggested that both promoters can drive enhanced green fluorescent protein (EGFP) specifically expressed in haemocytes. Further evidence clearly demonstrated that the transgenic silkworm exhibited a high level of EGFP signal in plasmatocytes, but not in other detected haemocyte types. Moreover, EGFP fluorescence signals were observed in the haematopoietic organ of both transgenic strains. Thus, two promoters that enable plasmatocytes to express genes of interest were confirmed in our study. It is expected that the results of this study will facilitate advances in our understanding of insect haematopoiesis and immunity in the silkworm, Bombyx mori.

“…The innate immunity consists of cellular immunity and humoral immunity. Cellular immunity mainly includes phagocytosis, encapsulation and formation of nodules, which are accomplished by hemocytes . Humoral immunity primarily includes the synthesis of antimicrobial peptides (AMPs) and the activation of phenoloxidase .…”

Section: Introductionmentioning

confidence: 99%

“…Cellular immunity mainly includes phagocytosis, encapsulation and formation of nodules, which are accomplished by hemocytes. [4][5][6] Humoral immunity primarily includes the synthesis of antimicrobial peptides (AMPs) and the activation of phenoloxidase. 7,8 These two different responses can be triggered by pattern recognition receptors (PRRs).…”

Section: Introductionmentioning

confidence: 99%

20‐Hydroxyecdysone enhances Immulectin‐1 mediated immune response against entomogenous fungus in Locusta migratoria

Han

Pest Management Science

et al. 2019

Background Entomogenous fungi are important factors in biological control, but innate immunity of insects restricts the efficiency of fungus infection. 20‐hydroxyecdysone (20E) is involved in regulating the immune response of insects. Our previous studies have revealed that 20E enhances the expression of antibacterial peptides in the worldwide pest Locusta migratoria. However, the mechanism by which 20E controls innate immunity against entomogenous fungi is still unknown. Results In the present study, based on the transcriptome of L. migratoria fat bodies challenged by 20E, immulectin‐1 (LmIML‐1) was screened and identified to be involved in modulating antifungal immunity. Spatio‐temporal expression analysis showed LmIML‐1 was highly expressed in the fifth instar nymph stage, and mainly distributed in the fat bodies and hemolymph. Both exogenous and endogenous 20E could increase the transcription of LmIML‐1. In contrast, transcription of LmIML‐1 did not increase when the 20E signal was blocked by RNAi of LmEcR (ecdysone receptor). The expressed recombinant protein rLmIML‐1 possessed agglutination activity and promoted the encapsulation. RNA interference of LmIML‐1 reduced the encapsulation of hemocytes, decreased the antifungal activity of plasma against Metarhizium anisopliae and accelerated the death of nymphs under the stress of entomogenous fungus. Meanwhile, 20E did not increase the antifungal activity with silence of LmIML‐1 in L. migratoria. Conclusion 20E enhances antifungal immunity by activating immulectin‐1 in L. migratoria. Our findings indicate a potential mechanism of 20E systematically regulating innate immune response to resist pathogens and provide a well‐defined molecular target for improving biological control. © 2019 Society of Chemical Industry