3-methylquercetin and Carcinoma--Hepatocellular

The immune checkpoint inhibitor (ICI) anti-PD-L1 monoclonal antibody can inhibit the progress of hepatocellular carcinoma (HCC). Epithelial-mesenchymal transformation (EMT) can promote tumor migration and the formation of immune-suppression microenvironment, which affects the therapeutic effect of ICI. Yin-yang-1 (YY1) is an important transcription factor regulating proliferation, migration and EMT of tumor cells. This work proposed a drug-development strategy that combined the regulation of YY1-mediated tumor progression with ICIs for the treatment of HCC.. We first studied the proteins that regulated YY1 expression by using pull-down, co-immunoprecipitation, and duo-link assay. The active compound regulating YY1 content was screened by virtual screening and cell-function assay. Isorhamnetin (ISO) and anti-PD-L1 antibody dual-functional mesoporous silica nanoparticles (HMSN-ISO@ProA-PD-L1 Ab) were prepared as an antitumor drug to play a synergistic anti-tumor role.. YY1 can specifically bind with the deubiquitination enzyme USP7. USP7 can prevent YY1 from ubiquitin-dependent degradation and stabilize YY1 expression, which can promote the proliferation, migration and EMT of HCC cells. Isorhamnetin (ISO) were screened out, which can target USP7 and promote YY1 ubiquitin-dependent degradation. The cell experiments revealed that the HMSN-ISO@ProA-PD-L1 Ab nanoparticles can specifically target tumor cells and play a role in the controlled release of ISO. HMSN-ISO@ProA-PD-L1 Ab nanoparticles inhibited the growth of Hepa1-6 transplanted tumors and the effect was better than that of PD-L1 Ab treatment group and ISO treatment group. HMSN-ISO@ProA-PD-L1 Ab nanoparticles also exerted a promising effect on reducing MDSC content in the tumor microenvironment and promoting T-cell infiltration in tumors.. The isorhamnetin and anti-PD-L1 antibody dual-functional nanoparticles can improve tumor immune microenvironment and inhibit YY1-mediated tumor progression. This study demonstrated the possibility of HCC treatment strategies based on inhibiting USP7-mediated YY1 deubiquitination combined with anti-PD-L1 monoclonal Ab.

Topics: Carcinoma, Hepatocellular; Cell Line, Tumor; Hereditary Sensory and Motor Neuropathy; Humans; Liver Neoplasms; Tumor Microenvironment; Ubiquitin-Specific Peptidase 7; Ubiquitins; YY1 Transcription Factor

Topics: Antineoplastic Agents, Phytogenic; Carcinoma, Hepatocellular; Cell Movement; Cell Proliferation; Cleome; Flavonoids; Glycosides; Hep G2 Cells; Humans; Liver Neoplasms; MicroRNAs; Molecular Structure; Quercetin; Tumor Suppressor Protein p53

Aflatoxin B(1) (AFB(1))-mediated hepatic damage is involved in production of AFB(1)-8,9-epoxide-bound DNA adducts and this is also affected by a pro-oxidant potential of the toxin. In this study we investigated the effects of quercetin on AFB(1)-treated HepG2 cells. We also examined the biochemical mechanisms associated with the effects of quercetin on AFB(1)-mediated liver damage in mice. Our results revealed that quercetin and isorhamnetin inhibit production of reactive oxygen species and cytotoxicity, and block the decrease of reduced glutathione (GSH) levels in AFB(1)-treated HepG2 cells. Isorhamnetin have inhibitory ability on lipid peroxidation stronger than quercetin in the cells. Oral supplementation with quercetin decreased serum lactate dehydrogenase levels, increased hepatic GSH levels and superoxide dismutase activity, and reduced lipid peroxidation in both the liver and kidney in AFB(1)-treated mice. However, quercetin did not show a significant reduction on serum levels of alkaline phosphate, alanine aminotransferase and aspartate aminotransferase that were increased in AFB(1)-treated mice. HPLC analysis revealed that quercetin in plasma is mainly present as glucoronides and/or sulfates of quercetin. Collectively, it is suggested that quercetin does not directly protect against AFB(1)-mediated liver damage in vivo, but exerts a partial role in promoting antioxidative defense systems and inhibiting lipid peroxidation.

Topics: Aflatoxin B1; Animals; Body Weight; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Survival; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Flavonols; Glutathione; Humans; Liver; Male; Malondialdehyde; Mice; Mice, Inbred ICR; Organ Size; Quercetin; Reactive Oxygen Species

Isorhamnetin, a flavonol aglycone, isolated from the traditional Chinese medicine Hippophae rhamnoides L., was investigated in its cytotoxicity and its influence on human hepatocellular carcinoma cells (BEL-7402). The cytotoxic effects of isorhamnetin showed dose- and time-dependency against BEL-7402 cells, with IC(50) equal to 74.4+/-1.13 microg ml(-1) after treatment with isorhamnetin for 72 h. Cytotoxicity of the flavonols on tumor cells depends on cellular accumulation of the drugs. The amount of isorhamnetin accumulated in BEL-7402 cells was assayed by high-performance liquid chromatography (HPLC) and showed that isorhamnetin could permeate the cell membrane into the cell. Staining with Hoechst 33258 showed fragmentation and condensation of chromatin in the cell treated with 50 microg ml(-1) isorhamnetin for 48 h. Flow cytometric analysis was performed to determine hypodiploid cells. The results of flow cytometry assay indicated that the percentage of hypodiploid BEL-7402 cells were 13.77+/-1.05% after 48 h treatment with 50 microg ml(-1) isorhamnetin. The treatment resulted in the appearance of a hypodiploid peak (sub-G(0)/G(1) peak), probably due to the presence of cells in apoptosis and apoptotic bodies with DNA content less than 2n. To our knowledge, this is the first report against human hepatocellular carcinoma cells (BEL-7402) of isorhamnetin.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Nucleus; Chromatography, High Pressure Liquid; Flavonols; G1 Phase; Hippophae; Humans; Liver Neoplasms; Quercetin