Differential expression of microRNAs in diapause and non-diapause gonads of Aspongopus chinensis Dallas (Hemiptera: Dinidoridae): implications for reproductive control.

Aspongopus chinensis Dallas, 1851 (Hemiptera: Dinidoridae), an edible and medicinal insect, usually found in China and Southeast Asia, offers substantial potential for various applications. The reproductive cycle of this particular insect occurs annually because of reproductive diapause, leading to inadequate utilization of available natural resources. Despite its considerable ecological importance, the precise mechanisms underlying diapause in A. chinensis are not yet well understood. In this study, we conducted an analysis of comparing the microRNA (miRNA) regulation in the diapause and non-diapause gonads of A. chinensis and identified 303 differentially expressed miRNAs, among which, compared with the diapause group, 76 miRNAs were upregulated and 227 miRNAs downregulated. The results, regarding the Enrichment analysis of miRNA-targeted genes, showed their involvement in several essential biological processes, such as lipid anabolism, energy metabolism, and gonadal growth. Interestingly, we observed that the ATP-binding cassette pathway is the only enriched pathway, demonstrating the capability of these targeted miRNAs to regulate the reproductive diapause of A. chinensis through the above essential pathway. The current study provided the role of gonadal miRNA expression in the control of reproductive diapause in A. chinensis, the specific regulatory mechanism behind this event remained unknown and needed more investigation.

Aspongopus chinensis Dallas induces pro-apoptotic and cell cycle arresting effects in hepatocellular carcinoma cells by modulating miRNA and mRNA expression.

Aspongopus chinensis Dallas is a traditional Chinese medicinal insect with several anticancer properties can inhibit cancer cell growth, by inhibiting cell division, autophagy and cell cycle. However, the precise therapeutics effects and mechanisms of this insect on liver cancer are still unknown. This study examined the inhibitory influence of A. chinensis on the proliferation of hepatocellular carcinoma (HCC) cells and explore the underlying mechanism using high-throughput sequencing. The results showed that A. chinensis substantially reduced the viability of Hep G2 cells. A total of 33 miRNAs were found to be upregulated, while 43 miRNAs were downregulated. Additionally, 754 mRNAs were upregulated and 863 mRNAs were downregulated. Significant enrichment of differentially expressed genes was observed in signaling pathways related to tumor cell growth, cell cycle regulation, and apoptosis. Differentially expressed miRNAs exhibited a targeting relationship with various target genes, including ARC, HSPA6, C11orf86, and others. Hence, cell cycle and apoptosis were identified by flow cytometry. These findings indicate that A. chinensis impeded cell cycle advancement, halted the cell cycle in the G0/G1 and S stages, and stimulated apoptosis. Finally, mouse experiments confirmed that A. chinensis significantly inhibits tumor growth in vivo. Therefore, our findings indicate that A. chinensis has a notable suppressive impact on the proliferation of HCC cells. The potential mechanism of action could involve the regulation of mRNA expression via miRNA, ultimately leading to cell cycle arrest and apoptosis. The results offer a scientific foundation for the advancement and application of A. chinensis in the management of HCC.

Aspongopus chinensis ach-miR-276a-3p induces breast cancer cell cycle arrest by targeting APPL2 to regulate the CDK2-Rb-E2F1 signaling pathway.

Breast cancer, the most common cancer, presents a significant challenge to the health and longevity of women. Aspongopus chinensis Dallas is an insect with known anti-breast cancer properties. However, the anti-breast cancer effects and underlying mechanisms have not been elucidated. Exogenous microRNAs (miRNAs), which are derived from plants and animals, have been revealed to have notable capacities for controlling the proliferation of cancerous cells. To elucidate the inhibitory effects of miRNAs derived from A. chinensis and the regulatory mechanism involved in the growth of breast cancer cells, miRNA sequencing was initially employed to screen for miRNAs both in A. chinensis hemolymph and decoction and in mouse serum and tumor tissue after decoction gavage. Subsequently, the experiments were performed to assess the suppressive effect of ach-miR-276a-3p, the miRNA screened out from a previous study, on the proliferation of MDA-MB-231 and MDA-MB-468 breast cancer cell lines in vitro and in vivo. Finally, the regulatory mechanism of ach-miR-276a-3p in MDA-MB-231 and MDA-MB-468 breast cancer cells was elucidated. The results demonstrated that ach-miR-276a-3p notably inhibited breast cancer cell proliferation, migration, colony formation, and invasion and induced cell cycle arrest at the G0/G1 phase. Moreover, the ach-miR-276a-3p mimics significantly reduced the tumor volume and weight in xenograft tumor mice. Furthermore, ach-miR-276a-3p could induce cell cycle arrest by targeting APPL2 and regulating the CDK2-Rb-E2F1 signaling pathway. In summary, ach-miR-276a-3p, derived from A. chinensis, has anti-breast cancer activity by targeting APPL2 and regulating the CDK2-Rb-E2F1 signaling pathway and can serve as a promising candidate anticancer agent.

Chromosomal-Level Genome Assembly of a True Bug, Aspongopus chinensis Dallas, 1851 (Hemiptera: Dinidoridae).

The true bug, Aspongopus chinensis Dallas, 1851 (Hemiptera: Dinidoridae), is a fascinating insect with prolonged diapause and medicinal properties but also a notorious pest. However, because of the lack of genomic resources, an in-depth understanding of its biological characteristics is lacking. Here, we report the first genome assembly of A. chinensis anchored to 10 pseudochromosomes, which was achieved by combining PacBio long reads and Hi-C sequencing data. This chromosome-level genome assembly was 1.55 Gb in size with a scaffold N50 of 156 Mb. The benchmarking universal single-copy ortholog (BUSCO) analysis of the assembly captured 96.6% of the BUSCO genes. A total of 686,888,052 bp of repeat sequences, 18,511 protein-coding genes, and 1,749 noncoding RNAs were annotated. By comparing the A. chinensis genome with that of 8 homologous insects and 2 model organisms, 213 rapidly evolving gene families were identified, including 83 expanded and 130 contracted gene families. The functional enrichment of Gene Ontology and KEGG pathways showed that the significantly expanded gene families were primarily involved in metabolism, immunity, detoxification, and DNA/RNA replication associated with stress responses. The data reported here shed light on the ecological adaptation of A. chinensis and further expanded our understanding of true bug evolution in general.

Chemical Composition and Antiproliferative Effects of a Methanol Extract of Aspongopus chinensis Dallas.

Natural products from insects can be potent sources for developing a variety of pharmaceutical products. Aspongopus chinensis Dallas has been used as a traditional Chinese medicine and there are several clinical evidences to support its anticancer activity. However, the anticancer active ingredients present in A. chinensis remain unidentified. In the present study, we investigated the anticancer effects of a methanol extract of A. chinensis (AME). Gas chromatography mass spectrometry was used to analyse the chemical composition of AME. The cell viability of MDA-MB-453 and HCC-1937 cells treated with different concentrations of AME was detected by MTT assay and the ratio of cells in different cell cycle phases was analysed by flow cytometry. The expression of genes associated with cell cycle was analysed by real-time PCR assay. The results showed that oleic acid (25.39%) and palmitic acid (21.798%) are the main anticancer compounds present in AME. There was a concentration-dependent decrease in the proliferation of MDA-MB-453 and HCC-1937 cells. Moreover, treatment with AME induced a S-phase arrest in the cells. Real-time PCR assay demonstrated that AME could significantly downregulate the expression of CDC20, AURKB, PLK1, CCNB2, and TOP2A mRNAs and upregulate the expression of GADD45A mRNA. We demonstrate that the methanol extract of A. chinensis could be a potential natural alternative or complementary therapy for breast cancer.

Antiproliferative and Proapoptotic Effects of a Protein Component Purified from Aspongopus chinensis Dallas on Cancer Cells In Vitro and In Vivo.

Aspongopus chinensis Dallas is used as a traditional Chinese medicine. In China, clinical evidence suggests that it has anticancer activity. However, the anticancer active components are not fully elucidated. In the present study, we purified an anticancer active component (named CHP) from A. chinensis. To gain a comprehensive insight into the protein components, shotgun proteomic analysis was conducted. The anticancer active protein band was cut from the sodium dodecyl sulphate-polyacrylamide gel electrophoresis gel and digested with trypsin to generate peptide mixture. The peptide fragments were then analysed by liquid chromatography tandem mass spectrometry; 18 proteins were identified. In addition, we evaluated the effects of CHP on the proliferation and apoptosis of two human gastric cancer cell lines (SGC-7901 and BGC-823). The cultured cells were treated with CHP at concentrations of 20, 30, and 40 µg/mL. Inhibition of cell growth was determined by the MTT assay. Hoechst 33258 staining was adopted to detect apoptosis morphologically. Apoptotic cells were quantified by Annexin V-FITC/propidium iodide staining and flow cytometry. Tumour growth was assessed by subcutaneous inoculation of 4T1 cells into BALB/c mice. There was a concentration- and time-dependent decrease in the proliferation of both cell lines at CHP concentrations of 20-40 µg/mL. Apoptotic characteristics, such as karyopyknotic pyknic hyperfluorescence bolus and nuclear fragmentation, were observed in both the cell lines by Hoechst 33258 staining. Flow cytometry showed that CHP induced significant (P < 0.01) concentration-dependent apoptosis of SGC-7901 cells. In vivo assay showed that CHP can partially inhibit tumour growth derived from 4T1 cells in vivo. The present study is the first to report that CHP in A. chinensis inhibits the proliferation of cancer cell lines via the suppression of cancer cell proliferation and acceleration of apoptosis.

The mitochondrial genome of Lasioderma serricorne (Coleoptera, Anobiidae).

In this study, the complete mitochondrial genome of the Lasioderma serricorne was sequenced and analysed. The mitochondrial genome is 14,476 bp long and contains 13 protein-coding genes, two rRNA genes, 22 tRNA genes, and two non-coding region. Twenty three genes were found to be encoded by the majority strand and the other 14 genes by minority strand, those similar to that of other insects. The nucleotide compositing of the majority strand is 39.74% of A, 11.20% of C, 40.47% of T, and 10.39% of G. The phylogenetic analysis by maximum-likelihood (ML) method revealed that the L. serricorne was close to Lasioderma redtenbacheri.

Acetamiprid inhibits testosterone synthesis by affecting the mitochondrial function and cytoplasmic adenosine triphosphate production in rat Leydig cells.

The insecticide acetamiprid is used to control noxious agricultural pests. However, it can cause mammalian toxicity. We evaluated the reproductive toxicity of acetamiprid in adult male Sprague Dawley rats. Rats were given oral acetamiprid alone or with vitamin E for 35 days. Rat plasma testosterone concentration and sperm quality decreased significantly as the levels of luteinizing hormone (LH) increased after exposure. At the same time, acetamiprid increased malondialdehyde and nitric oxide (NO) levels of Leydig cells. Further analysis showed that acetamiprid reduced the adenosine triphosphate (ATP) and cyclic adenosine monophosphate (cAMP) production of Leydig cells, but the expression of luteinizing hormone/choriogonadotropin receptor (LHCGR) and the activity of adenylyl cyclase were not changed. Acetamiprid exposure also significantly diminished protein levels of steroidogenic acute regulatory protein (STAR), hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase cluster (HSD3B), and cytochrome P450, family 11, subfamily a, polypeptide 1 (CYP11A1), and testicular mRNA levels, which are cAMP-dependent proteins that are essential for steroidogenesis. Electron microscopy indicated mitochondrial membrane damage in the Leydig cells of the testes of exposed rats. Vitamin E ameliorated the impairment of acetamiprid on Leydig cells. Our results indicate that acetamiprid causes oxidative stress and mitochondrial damage in Leydig cells and inhibits the synthesis of testicular ATP and cAMP. Acetamiprid disrupts subsequent testosterone biosynthesis by decreasing the rate of conversion of cholesterol to testosterone and by preventing cholesterol from entering the mitochondria within the Leydig cells. These effects caused reproductive damage to the rats.