Study on the Mechanism of CircDUSP16 Regulating the Proliferation and Apoptosis of Hepatocarcinoma Cells

Purpose: This study aims to explore the expression of circDUSP16 in liver cancer and its effect on the proliferation and apoptosis of liver cancer cells. Methods: Real-timePCR was used to measure the expression of circDUSP16, miR-136-5p, and YAP1 in HCC tissues and cells. MTT, colony analysis, and apoptosis analysis were performed to determine the progress of HCC cells. The relationship between circDUSP16, miR-136-5p, and YAP1 was verified by using the luciferase gene experiment. Results: The expression level of circDUSP16 in HCC tissue samples was significantly increased and can be used as an independent prognostic factor for the survival of HCC patients. Inhibition of circDUSP16 can inhibit HCC cell viability, colony formation, and invasion potential. Furthermore, inhibition of circDUSP16 can regulate the expression of YAP1 in the Hippo/YAP signaling pathway in HCC by targeting miR-136-5p, thereby affecting cell proliferation and apoptosis, and participating in the progression of HCC disease. Conclusion: The ectopic expression of circDUSP16 can regulate the expression of YAP1 by competitively binding to miR-136-5p, therefore participating in the progression of HCC, which can provide a new therapeutic target for the treatment of HCC.

Synthesis of deoxymannojirimycin using a Ruthenium-catalysed hydrogen borrowing reaction

<p>1-Deoxymannojirimycin (DMJ) has been investigated as a potential anti-cancer therapy due to its specific inhibition of class I α-mannosidase enzymes, which has been shown to trigger ER stress and the Unfolded Protein Response (UPR) pathway, leading to apoptosis in human hepatocarcinoma cells. Current methods for the synthesis of DMJ consist of multiple steps and often result in poor yields. The objectives of this research project were to develop a scale-up suitable synthesis of deoxymannojirimycin (DMJ), and to assess the feasibility of telescoping key-reactions to reduce the number of unit operations. Synthetic efforts focused on the key conversion of 1 to 2 have previously involved separate oxidation and reduction steps. In our laboratory; attempts to use hydrogen-borrowing chemistry had taken >48hr and not been achieved in high yield. The highlights of this work were that this conversion was ultimately realised in 95% yield in 24hr, and that the final deprotection of (2) could be telescoped into the process removing reaction-workup and chromatographic steps. The ruthenium catalyst used in the hydrogen borrowing reaction was found to be extremely air-sensitive, with reactions taking place in carefully prepared reaction vessels under an atmosphere of dry argon gas. The catalyst was also found to exhibit sensitivities to materials such as metal needles and polymer tubing, preventing sampling and monitoring of the reaction during synthesis. This study demonstrated that a one-pot synthesis is feasible,compressing the final steps in the synthesis of DMJ in excellent yield. The difficulty arises from the sensitive nature of the ruthenium catalyst, and the extreme care required in the preparation of the glassware and reagents used in synthesis. Many aspects of this development require further investigation, including the sampling, monitoring and quality control of each synthetic step.</p>

Synthesis of deoxymannojirimycin using a Ruthenium-catalysed hydrogen borrowing reaction

<p>1-Deoxymannojirimycin (DMJ) has been investigated as a potential anti-cancer therapy due to its specific inhibition of class I α-mannosidase enzymes, which has been shown to trigger ER stress and the Unfolded Protein Response (UPR) pathway, leading to apoptosis in human hepatocarcinoma cells. Current methods for the synthesis of DMJ consist of multiple steps and often result in poor yields. The objectives of this research project were to develop a scale-up suitable synthesis of deoxymannojirimycin (DMJ), and to assess the feasibility of telescoping key-reactions to reduce the number of unit operations. Synthetic efforts focused on the key conversion of 1 to 2 have previously involved separate oxidation and reduction steps. In our laboratory; attempts to use hydrogen-borrowing chemistry had taken >48hr and not been achieved in high yield. The highlights of this work were that this conversion was ultimately realised in 95% yield in 24hr, and that the final deprotection of (2) could be telescoped into the process removing reaction-workup and chromatographic steps. The ruthenium catalyst used in the hydrogen borrowing reaction was found to be extremely air-sensitive, with reactions taking place in carefully prepared reaction vessels under an atmosphere of dry argon gas. The catalyst was also found to exhibit sensitivities to materials such as metal needles and polymer tubing, preventing sampling and monitoring of the reaction during synthesis. This study demonstrated that a one-pot synthesis is feasible,compressing the final steps in the synthesis of DMJ in excellent yield. The difficulty arises from the sensitive nature of the ruthenium catalyst, and the extreme care required in the preparation of the glassware and reagents used in synthesis. Many aspects of this development require further investigation, including the sampling, monitoring and quality control of each synthetic step.</p>

Molecular Mechanism of Chrysin in Hepatocellular Carcinoma Treatment Based on Network Pharmacology and in Vitro Experiments

This study elucidated the potential molecular mechanism of chrysin in hepatocellular carcinoma (HCC) treatment using network pharmacology and in vitro experiments. Chrysin and candidate targets of HCC were obtained from the TCMSP and DrugBank databases, followed by mapping and screening of chrysin and HCC targets to identify the core targets of chrysin in HCC treatment. The interaction of chrysin and its targets, including CDK1, CDK5, as well as MMP9, were evaluated by molecular docking. The STRING database and Cytoscape (version 3.8.2) software were used to construct protein interactions and component-target networks of the core targets. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis of the core target genes were performed using the DAVID database. Network pharmacology results showed that chrysin treatment of HCC was mainly related to cell proliferation and cell cycle. Accordingly, the cell counting kit-8 method and flow cytometry were used to detect the cell viability and cell cycle of hepatocarcinoma cells HCCLM3 and BEL-7402 in vitro. A total of 142 compound targets of chrysin, 12,179 HCC-related targets, and 116 intersecting targets were screened. The first 20 GO biological annotations of 17 core targets and the first 20 KEGG pathways mainly involved cell proliferation and cell cycle. In vitro experiments showed that chrysin inhibits the proliferation of human hepatocarcinoma cells (HCCLM3 and BEL-7402) in a dose-dependent manner. Moreover, chrysin induced cell cycle arrest in HCCLM3 and BEL-7402 cells in the G2 phase, and the expression was downregulated of cyclin-dependent kinases (CDKs), CDK2 and CDK4. Chrysin can offset HCC mainly by regulating the cell cycle and inhibiting cell proliferation. The network pharmacology results were verified, providing the basis for further study on the mechanism of chrysin intervention in HCC.

Ficus septica, an ecosystem keystone species induced ROS-mediated cytotoxicity in HepG2 hepatocarcinoma cells

Ficus septica grows all around Indonesia, as one of the key species in various ecosystem types. This plant is a food resource and habitat for some animals. This study aimed to examine cytotoxic activity of F. septica extract in HepG2 cells. The leaves powder was macerated using ethanol 96%. Cytotoxic activity was evaluated by MTT assay. The determination of cell cycle profile and reactive oxygen species (ROS) were done by flow cytometry. The extract inhibited the growth of HepG2 and Vero cells with an IC50 of 50.9 and 286.2 µg/mL, while doxorubicin 0.8 and 12.8 µg/mL. The selectivity index of the extract and doxorubicin was 5.6 and 16 respectively. The extract triggered cell cycle arrest in HepG2 at the G0/G1 phase, whereas doxorubicin in the S phase. The extract and doxorubicin significantly increased intracellular ROS in HepG2, but not in normal Vero cells. In conclusion, our findings suggest that F. septica induced cytotoxicity in HepG2 cells is mediated by excessive ROS generation leading to oxidative stress.

YRDC Mediates the Resistance of Lenvatinib in Hepatocarcinoma Cells via Modulating the Translation of KRAS

Lenvatinib is the latest and promising agent that has demonstrated a significant improvement of progression-free survival in advanced hepatocellular carcinoma (HCC). However, resistance emerges soon after initial treatment, limiting the clinical benefits of lenvatinib. Therefore, understanding the mechanism of resistance is necessary for improving lenvatinib efficacy. YRDC promotes the proliferation of hepatocarcinoma cells via regulating the activity of the RAS/RAF/MEK/ERK pathway, which was the primary pathway of the anticancer effect of lenvatinib. The purpose of this study is to investigate whether YRDC modulates the sensitivity of lenvatinib in hepatocarcinoma cells. Using the CCK-8 cell viability assay, wound-healing assay and clone formation assay in cell models, and xenograft assay in null mouse, we demonstrated that Huh7 cells with YRDC knockdown showed decreased susceptibility to lenvatinib than their control cells. Furthermore, we found that lenvatinib inhibited the expression of YRDC in a time-dependent manner. This effect may aggravate resistance to lenvatinib in hepatocarcinoma cells and may be an underlying cause of resistance, which emerges soon after lenvatinib initial treatment. To investigate how YRDC modulates the sensitivity of lenvatinib, we assessed the effect of tRNA with different t6A levels on the translation of the KRAS gene by in vitro rabbit reticulocyte translation system and measured the expression levels of the KRAS gene by western blot together with qPCR. We found that YRDC regulates the protein translation of KRAS in cell models, and the tRNA with low t6A modification level reduces the translation of the KRAS in the in vitro translation system. These results suggested that YRDC mediates the resistance of lenvatinib in hepatocarcinoma cells via modulating the translation of the KRAS. In this study, YRDC was confirmed to be a potential novel predictive biomarker of lenvatinib sensitivity in HCC.