Anti-hepatocellular Carcinoma Research Articles

Potential anti-hepatocellular carcinoma properties and mechanisms of action of clerodane diterpenes isolated from Polyalthia longifolia seeds

Diterpenes are secondary metabolites that have attracted much attention due to their potential biological activities including anti-cancer potential. The aim of the current study is to assess the anticancer potential of the six known clerodane diterpenes (1–6) isolated from Polyalthia longifolia seeds and their underlying molecular mechanisms. These compounds were evaluated for their cytotoxicity in vitro by using MTT assays. The “two-phase model” with NDEA and PB ad libitum was used for induction of HCC and sorafenib was used as the standard drug. Prophylactic studies were carried out for compounds 4/6 at both low (5 mg/kg b.w) and high (10 mg/kg b.w) doses. Based on the MTT assay results, the two best compounds, 4 and 6, were selected for in vivo studies. The results showed that treatment with compound 4/6 significantly restored the changes in biochemical parameters and liver morphology observed in (NDEA + PB)-induced HCC rats. Additionally, the docking studies showed that compound 4/6 interacted with several key proteins such as MDM2, TNF-α, FAK, thereby inhibiting these proteins and reversing the negative impacts of NDEA. In conclusion, our results suggested that compounds 4 and 6 are potential therapeutic agents for HCC, mostly due to their ability to control typical cancer pathways.

Fuzheng Xiaozheng prescription exerts anti-hepatocellular carcinoma effects by improving lipid and glucose metabolisms via regulating circRNA-miRNA-mRNA networks

BackgroundHepatocellular carcinoma (HCC) is a major threat to human health due to its high lethality. Our previous studies suggested that Fuzheng Xiaozheng prescription (FZXZP), an effective Chinese medicine, demonstrated significant suppressive effects on HCC. However, its underlying mechanism remains largely unclear. PurposeThis study aimed to investigate the anti-HCC mechanisms of FZXZP from transcriptomic sequencing based on a holistic perspective. MethodsRat HCC model was induced by diethylnitrosamine, and then the model was administered with two doses of FZXZP, high and low. Sodium demethylcantharidate was used as a positive control. Subsequently, microarrays of circRNA, miRNA and mRNA were performed on the blank, model, high and low dose groups, respectively, and the competitive binding mechanisms among them were further analyzed by bioinformatics. Then, the circRNA-miRNA-mRNA networks were constructed to mine the targeted-RNAs of FZXZP in HCC, as well as to explore their potential regulatory mechanisms. Finally, functions and pathways of the FZXZP targeted genes in rat HCC were annotated with GO and KEGG, and qRT-PCR was performed to validate the accuracy of the above analyses in this study. ResultsThe results showed that FZXZP significantly inhibited the development and progression of HCC in rats, improved the pathological conditions and suppressed the proliferation of HCC cells. Subsequently, after a series of screening, the competing endogenous RNA networks (circRNA-miRNA-mRNA), consisting of 2 circRNAs, 7 miRNAs and 104 mRNAs, were finally established. KEGG and GO analyses of the networks revealed that lipid metabolism related pathways, such as fatty acid metabolism, bile secretion and PPAR pathway, were significantly enriched. In the further hubgene network analysis, in addition to lipid metabolism, aberrant glucose metabolism was found to be ameliorated by G6pc and Pklr in hubgenes. Finally, the qRT-PCR analyses confirmed that the expression tendencies of the above targeted genes were correct and believable in transcriptomic sequencings, and qRT-PCR results of the genes closely related to proliferation, invasion and apoptosis of HCC also indicated the inhibitory effects of FZXZP on HCC obviously. ConclusionFZXZP demonstrated significant anti-HCC effects through improving lipid and glucose metabolism, restoring the metabolic homeostasis of the liver via circRNA-miRNA-mRNA networks.

Design, Synthesis, and Biological Evaluation of Novel Evodiamine Derivatives as Potential Antihepatocellular Carcinoma Agents.

Evodiamine has many biological activities. Herein, we synthesize 23 disubstituted derivatives of N14-phenyl or the E-ring of evodiamine and conduct systematic structure-activity relationship studies. In vitro antiproliferative activity indicated that compounds F-3 and F-4 dramatically inhibited the proliferation of Huh7 (IC50 = 0.05 or 0.04 μM, respectively) and SK-Hep-1 (IC50 = 0.07 or 0.06 μM, respectively) cells. Furthermore, compounds F-3 and F-4 could double inhibit topoisomerases I and II, inhibit invasion and migration, block the cell cycle to the G2/M stage, and induce apoptosis as well. Additionally, compounds F-3 and F-4 could also inhibit the activation of HSC-T6 and reduce the secretion of collagen type I to slow down the progression of liver fibrosis. Most importantly, compound F-4 (TGI = 60.36%) inhibited tumor growth more significantly than the positive drug sorafenib. To sum up, compound F-4 has excellent potential as a strong candidate for the therapy of hepatocellular carcinoma.

Targeted Anti-Hepatocellular Carcinoma Research of Targeted Peptides Combined with Drug-Loaded Cell-Derived Microparticles.

To conduct an anti-tumor research by using targeted drug-loaded cell-derived microparticles to target the tumor microenvironment and enhance NK cell killing function. In this experiment, we obtained HepG2 tumor cell-derived microparticles by physical extrusion, high speed centrifugation and filtration, modified the hepatocellular carcinoma targeting peptide SP94 on the surface of microparticles and encapsulated the TGF-β inhibitor SB505124. Finally we validated and analyzed whether the new drug delivery system can target to tumor site and enhance the anti-tumor function of NK cells. This type of novel targeted cell-derived microparticles drug delivery system will provide a novel idea for tumor immunotherapy.

Mechanisms underlying the activity of paederus in hepatocellular carcinoma: A network pharmacology and in vitro validation approach

Paederus, belonging to the Coleoptera and Staphylinidae, can prevent cell mitosis and inhibit tumors. However, few studies exist on the mechanism of Paederus on inhibiting the growth of tumors. Consequently, the present study used network pharmacology, molecular docking and in vitro experiments explore the anti-Hepatocellular Carcinoma (HCC) mechanism of Paederus. MethodsNetwork pharmacology was used to predict the potential mechanisms through which Paederus acts against HCC. Therefore, the active compounds of Paederus were retrieved from the CNKI, Wanfang, PubMed and Web of Science databases. The targets for the active compounds were obtained from the Targetnet database, then the common targets and signaling pathways were obtained by Protein-protein interaction (PPI) network analysis, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. Thereafter, the compound-target and target-pathway networks were constructed. Finally, in vitro experiments using the SMMC-7721 HCC cells were used to further verify the predicted anti-HCC mechanism of Paederus. ResultsThe study screened 40 active compounds from Paederus, and 26 targets related to the anti-HCC mechanism of Paederus were predicted through network pharmacology. These targets were involved in the Nod-like receptor, MAPK, PI3K- Akt, TNF and AMPK signaling pathways. In addition, cell experiments showed that Paederus reduce the proliferation rate of SMMC-7721 HCC cells. Moreover, Paederus could down-regulated the expressions of the HSP90AA1, IRAK-4 and CTSB proteins in the Nod-like receptor signaling pathways in SMMC-7721 HCC cells. ConclusionThe study revealed the potential mechanism through which Paederus acts against HCC by combining network pharmacology, molecular docking, and experimental verification.

6-Amino-2,4,5-trimethylpyridin-3-ol and 2-amino-4,6-dimethylpyrimidin-5-ol derivatives as selective fibroblast growth factor receptor 4 inhibitors: design, synthesis, molecular docking, and anti-hepatocellular carcinoma efficacy evaluation

A novel series of aminotrimethylpyridinol and aminodimethylpyrimidinol derivatives were designed and synthesised for FGFR4 inhibitors. Structure-activity relationship on the FGFR4 inhibitory activity of the new compounds was clearly elucidated by an intensive molecular docking study. Anti-cancer activity of the compounds was evaluated using hepatocellular carcinoma (HCC) cell lines and a chick chorioallantoic membrane (CAM) tumour model. Compound 6O showed FGFR4 inhibitory activity over FGFR1 − 3. Compared to the positive control BLU9931, compound 6O exhibited at least 8 times higher FGFR4 selectivity. Strong anti-proliferative activity of compound 6O was observed against Hep3B, an HCC cell line which was a much more sensitive cell line to BLU9931. In vivo anti-tumour activity of compound 6O against Hep3B-xenografted CAM tumour model was almost similar to BLU9931. Overall, compound 6O, a novel derivative of aminodimethylpyrimidinol, was a selective FGFR4 kinase inhibitor blocking HCC tumour growth.

Antiproliferative Activity of Some Newly Synthesized Substituted Nicotinamides Candidates Using Pyridine-2(1H) thione Derivatives as Synthon.

Some new pyridinethione and thienopyridine derivatives have been synthesized and evaluated for their antiproliferative activity using the MTT assay. Nicotinamide derivatives 3 have been synthesized and used for the preparation of new condensed thieno [2,3-b]pyridines by their reactions with active halo compounds. Finally the synthesized thienopyridine underwent ring closure whenever possible through boiling in a solution of sodium ethoxide. The antiproliferative evaluation against (HCT-116, HepG-2, and MCF-7) human cancer cells and one human healthy cell line (BJ-1) revealed that compounds 3b, 4c–5d, 7b–12a, 10d, and 13b have interesting antitumor activity specifically as antihepatocellular and anticolon cellular carcinoma agents. Besides, the docking results for most active derivatives were in agreement with the in vitro antitumor results.

Effects of Ganfule capsule on microbial and metabolic profiles in anti-hepatocellular carcinoma.

Based on the gut microbiota and plasma metabolites, the underlying mechanism was analysed for the anti-hepatocellular carcinoma (HCC) effects of Ganfule capsule (GFL) in the study. The UPLC-Q-TOF/MS results showed that 13key compounds were identified in GFL and the major active ingredients included amygdalin, saikosaponin A, astragaloside I, etc. The nude mice received HepG2 injection, and GFL showed lower volume and weight of the tumour. In addition, the apoptosis proteins (Bax and Bcl2) were altered in response to GFL treatment, and apoptosis cells were increased, indicating an anti-HCC effect. Interestingly, 16S rDNA results showed that GFL treatment improved gut microbiota diversity and compositions, especially for the beneficial bacteria, such as Bacilli, Lactobacillales, Lactobacillus, Lactobacillaceae, Firmicutes, Lactobacillus_reuteri and Lactobacillus_gasseri. Metabonomics further identified 426metabolites and 343metabolites variation in the positive and negative ion modes after GFL treatment, which might be associated with amino acid, lipid metabolism and carbohydrate metabolism pathways, indicating these metabolites might involve in the protective role of GFL in HCC. Correlation analysis showed a significant relationship between gut microbiota and plasma metabolites. In conclusion, GFL exerted an anti-HCC effect in the nude murine model, which might be associated with microbial and metabolic improvements. This study is the first to report the anti-HCC effect of GFL associated with gut microbiota and plasma metabolites. GFL may improve the gut microbiota structure, such as increasing probiotics - Lactobacillus. It also provides a new strategy for the scientific demonstration of the modernization of traditional Chinese medicine.

Dihydromyricetin from Ampelopsis grossedentata and its derivatives: Structural characterization and anti-hepatocellular carcinoma activity

Dihydromyricetin (DMY) was efficiently extracted from Ampelopsis grossedentata and modified by esterification to obtain two derivatives, which were named as benzoylated derivative (BZ-DMY) and acetylated derivative (AC-DMY). The physicochemical characterizations of BZ-DMY and AC-DMY were investigated by TLC, SEM, HPLC, FT-IR, NMR and MS analysis. Simultaneously, the anti-hepatocellular carcinoma activity was determined by cell experiment to investigate their structure-activity relationship. The results indicated that esterification of DMY were successfully modified by benzoyl and acetyl. The substitute positions were C-3, C-5, C-7, C-3′, C-4′ and C-5′ respectively inferred from NMR analysis. In the experiment of anti-hepatocellular carcinoma activity, two novel derivatives showed better anti-tumor ability than DMY. Therefore, it can be concluded that esterified DMY could significantly induce HepG2 cells apoptosis and improve the anti-tumor activity of DMY. In summary, this research could not only enhance the development and utilization of Ampelopsis grossedentata, but also develop these two derivatives (BZ-DMY and AC-DMY) as potential anti-hepatocellular carcinoma therapeutic drugs.

The nature compound dehydrocrenatidine exerts potent antihepatocellular carcinoma by destroying mitochondrial complexes in vitro and in vivo.

Cumulative evidence indicates that mitochondria dysfunction plays an important role in tumour treatment. Given the limited efficacy and toxicity of current mitochondria-targeted drugs, research into effective mitochondria-targeted anticancer agentsremains an irresistible general trend. In this study, it was found that dehydrocrenatidine (DEC), a β-carbolin alkaloid isolated from Picrasma quassiodes, displays a promising growth inhibitory effect in vitro and in vivo by inducing apoptosis of hepatocellular carcinoma (HCC) cells. Mechanistically, we provided that the possible target of DEC against HCC cells was determined by isobaric labels for relative and absolute quantification assay and validated them using further experiments. The results suggested that DEC can target and regulate the function of mitochondrial complexes I, III and IV, affecting oxidative phosphorylation and ultimately leading to mitochondrial dysfunction to exert its anti-HCC effects. In addition, the combination of DEC and sorafenib showed a synergistic effect and was also associated with mitochondrial dysfunction. Importantly, DEC did not show significant toxicity in mice. This study provided a new insight into underlying mechanisms in DEC-treated HCC cells, suggesting that DEC might be a mitochondrial targeting lead compound.

CRHBP is degraded via autophagy and exerts anti-hepatocellular carcinoma effects by reducing cyclin B2 expression and dissociating cyclin B2-CDK1 complex.

Autophagy is the predominant self-eating catabolic pathway activated in response to nutrient starvation and hypoxia within the microenvironment of varied malignancies, including hepatocellular carcinoma (HCC). SQSTM1/p62 links its cargos to autophagosomes for degradation, and reportedly acts as a contributor for hepatocarcinogenesis. Five GEO gene microarrays identified corticotropin releasing hormone (CRH) binding protein (CRHBP) as a significantly downregulated gene in HCC (log2 Fold change < -3 and p < 0.001), and an earlier human interactome study indicated that CRHBP may interact with p62. This study aimed to explore (1) the role of CRHBP in HCC development, and (2) whether p62-mediated autophagy was responsible for low CRHBP expression within HCC tissue. Following functional experiments first revealed an anti-proliferative, anti-metastatic, and anti-angiogenic role of CRHBP in HCC cells (Huh-7, Li-7 and HCCLM3) and xenografts. CRHBP negatively regulated cyclin B2 expression, and dissociated cyclin B2-CDK1 complex in HCC cells, thereby leading to cell cycle arrest at G2 phase. To simulate HCC microenvironment in vitro, Huh-7 cells were incubated in Earle's Balanced Salt Solution (nutrient starvation) or exposed to 1% O2 (hypoxic exposure). In addition to activating autophagy, nutrient starvation and hypoxic exposure also induced CRHBP degradation. Interestingly, CRHBP was demonstrated as a novel cargo targeted by p62 for degradation in autophagosomes. Blocking autophagy with 3-MA, chloroquine or siSQSTM1 prevented CRHBP degradation in HCC cells. Collectively, our study uncovers a role for CRHBP in retarding HCC development, reducing cyclin B2 expression and impairing cyclin B2-CDK1 interaction. CRHBP downregulation in HCC may attribute to p62-mediated autophagy.

Synthesis and Anti-Hepatoma Activities of U12 Derivatives Arresting G0/G1 Phase and Inducing Apoptosis by PI3K/AKT/mTOR Pathway.

Ursodeoxycholic acid (UDCA) is a first-line clinical drug for the treatment of liver diseases. U12, a derivative of UDCA, showed effective anti-hepatoma activities in previous works. However, the low polarity and large doses limited the druglikeness of U12. In this study, the structural modification and optimization of U12 were further investigated and twelve U12 derivatives were synthesized by substitution, esterification and amidation reactions. The evaluation of the cytotoxicity of synthetic derivatives against hepatoma cell lines (HepG2) indicated that U12-I, U12a-d and U12h showed more effective cytotoxic effects on the growth of HepG2 cells than U12, and the preliminary structure–activity relationship was discussed. Among them, U12a exhibited the most potent anti-hepatocellular carcinoma activity. Mechanism studies indicated that U12a inhibited HepG2 cell proliferation by arresting the G0/G1 phase, and suppressed the activation of the PI3K/AKT/mTOR pathway. Further studies showed that U12a induced HepG2 cells apoptosis through activating the caspase signaling pathway. Furthermore, U12a evidently inhibits the growth of HepG2-derived tumor xenografts in vivo without observable adverse effects. Thus, U12a might be considered as a promising candidate for the treatment of hepatocellular carcinoma.

Anti-Hepatocellular Carcinoma Effect and Molecular Mechanism of the Estrogen Signaling Pathway.

There are significant gender differences in the incidence and mortality of hepatocellular carcinoma (HCC). Compared with men, the incidence and mortality of HCC in women are relatively low. The estrogen signaling pathway, composed of estrogen and estrogen receptors, has been postulated to have a protective effect on the occurrence and development of HCC. There have been multiple studies that have supported anti-HCC effects of the estrogen signaling pathways, including direct and indirect pathways such as genomic pathways, rapid transduction pathways, non-coding RNA, tumor microenvironment, estrogen metabolites, and inhibition of hepatitis infection and replication. Based on the evidence of an anti-HCC effect of the estrogen signaling pathway, a number of strategies have been investigated to determine the potential therapeutic effect. These have included estrogen replacement therapy, targeting the estrogen receptor, key molecules, inflammatory mediators, and regulatory pathways of the estrogen signaling pathway. In this review, we have systematically summarized the latest developments in the complex functions and molecular mechanisms of the estrogen signaling pathway in liver cancer. Furthermore, we have highlighted the potential targets of treatment strategies based on the estrogen signaling pathway in the treatment of liver cancer and the principal obstacles currently encountered for future investigation.

Mollugin induced oxidative DNA damage via up-regulating ROS that caused cell cycle arrest in hepatoma cells

Mollugin has been proven to have anti-tumor activity. However, its potential anti-tumor mechanism remains to be fully elaborated. Herein, we investigated the growth inhibition of HepG2 cells, as well as the anti-tumor effect of mollugin and its molecular mechanism on H22-tumor bearing mice. In vitro, mollugin was shown to have a strong inhibitory effect on HepG2 cells in a concentration-dependent manner. Mollugin induced S-phase arrest of HepG2 cells, and increased intracellular reactive oxygen species (ROS) levels. Comet assay demonstrated that mollugin induced DNA damage in HepG2 cells, as well as an increase in the expression of p-H2AX. In addition, mollugin induced changes in cyclin A2 and CDK2. However, the addition of antioxidant glutathione (GSH) was able to reverse the effect of mollugin. In vivo, mollugin significantly inhibited tumor growth and reduced the tendency of tumor volume growth in mice. The tumor cell density was found to be decreased in the administration group, and the content of ROS in the tumor tissue significantly increased. The expression of p-H2AX, cyclin A2 and CDK2 were consistent with in vitro results. Mollugin demonstrated anti-hepatocellular carcinoma activity in vitro and in vivo, and its anti-hepatocellular carcinoma activity was found to be related to DNA damage and cell cycle arrest induced by excessive ROS production in cells.

The inhibitory effect of Periplaneta americana L. on hepatocellular carcinoma: Explore the anti-hepatocellular carcinoma active site and its mechanism of action

Ethnopharmacological relevanceThe American cockroach (Periplaneta americana L.) belongs to the family Blattidae, order Blattodea, and class Insecta. Its medicinal history in China spans thousands of years. In recent years, the anti-tumour activity of American cockroach has gradually attracted the attention of researchers and has a good application prospect in the treatment of tumours. Periplaneta americana has been found to contain proteins, peptides, amino acids and nucleosides. Pharmacological studies have shown that P. americana has anti-tumour, tissue repair, immunoregulatory and other activities. In this study, we investigated the chemical composition and mechanism of action of its active site against hepatocellular carcinoma. Materials and methodsWe adopted ultra-performance liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry (UPLC–Q-Orbitrap HRMS), measuring the accurate relative molecular mass, fragment ion peak, chromatographic retention time and reference substance information of the compound obtained by HRMS, to identify the chemical components of the anti-hepatocellular carcinoma (HCC) active site of P. americana based on data from relevant literature. We used western blotting (WB) to detect the expression levels of phosphoinositide 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt) and Akt in the PI3K/Akt pathway and further study the molecular mechanism of the active site of P. americana against HCC. ResultsUPLC–Q-Orbitrap HRMS identified 35 compounds from the active site of P. americana. Of these, 10 were amino acids, 1 was an alkaloid, 6 were nucleosides and their bases, 4 were dipeptides and cyclic dipeptides, 8 were organic acids, 2 were isoflavones and 4 were other compounds; 8 of these compounds were confirmed by comparison with the reference substance. The WB results showed that the relative expression levels of PI3K and p-Akt protein in the active site of P. americana in the medium-dose (concentration, 0.15624 mg⋅mL−1) and high-dose (concentration, 0.31250 mg⋅mL−1) experimental groups were significantly reduced compared with the blank control group (P < 0.05 or P < 0.01), whereas the expression level of Akt protein did not significantly change amongst the groups (P > 0.05). ConclusionThis study found that the anti-HCC active site of P. americana is composed of multiple components that can reduce the relative expression of PI3K and p-Akt protein. It exerts its anti-HCC effect by regulating the PI3K/Akt pathway.

The Pyroptosis-Related Gene Signature Predicts the Prognosis of Hepatocellular Carcinoma.

Objective: Hepatocellular carcinoma (HCC) is a genetically and phenotypically heterogeneous tumor, and the prediction of its prognosis remains a challenge. In the past decade, studies elucidating the mechanisms that induce tumor cell pyroptosis has rapidly increased. The elucidation of their mechanisms is essential for the clinical development optimal application of anti-hepatocellular carcinoma therapeutics. Methods: Based on the different expression profiles of pyroptosis-related genes in HCC, we constructed a LASSO Cox regression pyroptosis-related genes signature that could more accurately predict the prognosis of HCC patients. Results: We identified seven pyroptosis-related genes signature (BAK1, CHMP4B, GSDMC, NLRP6, NOD2, PLCG1, SCAF11) in predicting the prognosis of HCC patients. Kaplan Meier survival analysis showed that the pyroptosis-related high-risk gene signature was associated with poor prognosis HCC patients. Moreover, the pyroptosis-related genes signature performed well in the survival analysis and ICGC validation group. The hybrid nomogram and calibration curve further demonstrated their feasibility and accuracy for predicting the prognosis of HCC patients. Meanwhile, the evaluation revealed that our novel signature predicted the prognosis of HCC patients more accurately than traditional clinicopathological features. GSEA analysis further revealed the novel signature associated mechanisms of immunity response in high-risk groups. Moreover, analysis of immune cell subsets with relevant functions revealed significant differences in aDCs, APC co-stimulation, CCR, check-point, iDCs, Macrophages, MHC class-I, Treg, and type II INF response between high- and low-risk groups. Finally, the expression of Immune checkpoints was enhanced in high-risk group, and m6A-related modifications were expressed differently between low- and high-risk groups. Conclusion: The novel pyroptosis-related genes signature can predict the prognosis of patients with HCC and insight into new cell death targeted therapies.

Retraction: Synthesis of xanthone derivatives and anti-hepatocellular carcinoma potency evaluation: induced apoptosis.

Retraction of ‘Synthesis of xanthone derivatives and anti-hepatocellular carcinoma potency evaluation: induced apoptosis’ by Jie Liu et al., RSC Adv., 2019, 9, 40781–40791, https://doi.org/10.1039/C9RA06408G.

Synthesis, characterization, and anti-hepatocellular carcinoma effect of glycyrrhizin-coupled bovine serum albumin-loaded luteolin nanoparticles

Background: Luteolin (Lut) is a natural flavonoid with low water solubility. Many studies have revealed that its antitumor effect is more understandable. Objectives: Synthesis of glycyrrhizin-coupled bovine serum albumin-loaded Lut nanoparticles (GL-BSA-Lut-Nps) to progress the water solubility, anticancer effect, liver targeting, cycle arrest, induction apoptosis, and regulating the metabolism of Lut. Materials and Methods: The activity screening of GL-BSA-Lut-Nps on Hepatocellular Carcinoma Bel-7402 cells was spotted by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, cell cycle, and apoptosis were measured by flow cytometry. The solubility of GL-BSA-Lut-Nps was evaluated by ultraviolet spectrophotometer. Fluorescein isothiocyanate was employed to label the drug, and the fluorescence intensity of cells after drug uptake was detected under a fluorescence microscope to sense the targeting of GL-BSA-Lut-Nps to tumor cells. The differences of metabolites between Bel-7402 cells treated with GL-BSA-Lut-Nps and the control group were considered by 1 hydrogen-nuclear magnetic resonance metabolomics. Results: The results presented that the GL-BSA-Lut-Nps prepared by solvent removal method had good anti-tumor activity and water solubility in vitro and the No. 4 Lut nanoparticles (GL-BSA-Lut-Nps-4) screened by the MTT method had the best effect. The IC50 of the GL-BSA-Lut-Nps-4 on the Bel-7402 cell inhibition test was 1.999 ± 0.880 mg/mL. The results of cell cycle and apoptosis displayed that the anticancer effect of the prescription is more palpable. The results of the fluorescence original method are proposed to confirm that the experimentally created GL-BSA-Lut-Nps-4 have a liver targeting effect. The study of metabolomics further clarified the metabolic regulation effect of GL-BSA-Lut-Nps-4 on Bel-7402 cells. Conclusion: It delivers a theoretical basis for the development of new high-efficiency and low-toxicity traditional Chinese medicine preparations for liver cancer.

Nanobubble-based anti-hepatocellular carcinoma therapy combining immune check inhibitors and sonodynamic therapy.

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide, posing a global threat to human healthcare, and current approved treatment strategies do not produce satisfactory outcomes. Here, nanobubbles (NBs) were prepared that carried Immune Check Inhibitors (ICIs), PD-L1 antibody (PD-L1 Ab) and sonodynamic agent CHLORIN E6 (Ce6); the anti-cancer properties of these NBs were analyzed from the point of view of immune and sonodynamic therapies. The PD-L1 Ab/Ce6-NBs could inhibit tumor growth through regulating reactive oxygen species (ROS) production, apoptosis, and most importantly, the function of associated immunocytes, including natural killer cells and lymphocytes. The tumor tissues highly expressed markers of immunogenic tumor cell death (ICD) in which the expression of calreticulin (CRT) and ICD-related immune cytokines (CD80, CD86, INF-γ, and IL-2) were increased in PD-L1 Ab/Ce6-NB treated mice. PD-L1 Ab/Ce6-NBs also promoted murine spleen lymphocyte proliferation and cytotoxic activity, as well as CD8+ T cell infiltration in the tumor tissues, and downregulation of the PD-L1 protein and mRNA expression. Furthermore, Bax expression was increased and Bcl-2 was inhibited at the mRNA and protein levels in a murine subcutaneous transplanted tumor model. These results indicate that PD-L1 Ab/Ce6-NBs can induce ROS-dependent ICD to further boost anti-cancer immune responses under the action of targeting the PD-1/PD-L1 immune check point in the tumor microenvironment as a promising therapeutic agent for HCC.

Anti-hepatocellular carcinoma efficacy of Fuzheng Xiaozheng prescription and its interventional mechanism studies

Ethnopharmacological relevanceFuzheng Xiaozheng prescription (FZXZP), a traditional Chinese medicine, which was derived from the famous decoction, Sanjiasan, in the book of “Wenyilun” in Ming dynasty. Due to its function of invigorating the circulation of blood in Chinese medicine, it was usually used for treating the liver cirrhosis, hepatocellular carcinoma (HCC), etc. Clinical application found that FZXZP exhibited satisfactory therapeutic effects in HCC treatments. However, we still know little about the underlying mechanisms. Aim of studyIn this study, we aim to gain a deeper insight into the inhibiting effects of FZXZP on HCC rats and preliminarily elucidate the underlying intervention effects. Materials and methodsTwo doses of FZXZP were adopted to evaluate the therapeutic effects on rat HCC, and then the intervention effects were evaluated from different aspects. High performance liquid chromatography (HPLC) was used for the active compounds prediction in FZXZP. Finally, the mRNA-Seq was conducted to reveal the intervention mechanisms and the mechanisms were further validated by quantitative Real-time PCR (qRT-PCR) and lipid contents analyses. ResultsThe results showed that FZXZP significantly alleviated the serum biochemical indicators and improved the pathological characteristics of HCC rats. Mechanistically, FZXZP could regulate some lipid related metabolisms, including arachidonic acid, linoleic acid and retinol, as well as improving the steroid hormone biosynthesis, to improve the inflammatory statuses and restoring ability of HCC livers, and these were further confirmed by our following analyses on serum lipid contents and cytokine expressions. In addition, FZXZP could also negatively regulate four extracellular growth factors which could result in the blocking of two cancer-related signaling pathways, Ras/MAPK and Ras/PI3K-Akt. Conclusion: Our results suggested that FZXZP demonstrated significant inhibiting effects on rat HCC progresses, and these may be realized by improving the inflammatory statuses and blocking the Ras/MAPK and Ras/PI3K-Akt signaling pathways.