Integrative analysis of the molecular signature of target genes involved in the antitumor effects of cantharidin on hepatocellular carcinoma.

BACKGROUND: Cantharidin (CTD) is the active ingredient of Chinese medicine, which has been traditionally used in multiple cancers treatment, especially in hepatocellular carcinoma (HCC). However, a comprehensive analysis of the CTD-related molecular mechanism is still necessary to understand its functions in HCC treatment. This study aimed to reveal the novel molecular targets and regulatory networks of CTD in HCC. METHODS: A model of H22 tumour-bearing mice was constructed, and the function of CTD in tumour growth was evaluated. An integrated approach of CTD associated transcriptional profiling and biological systems analysis was used to identify key regulators involved in antitumour pathways. The identified differential expression patterns were supported by the results of Gene Ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyse, and by protein-protein interaction (PPI) network construction. The relationships between gene expression and tumour immunity were evaluated using Tumour Immune Estimation Resource (TIMER). Prognostic value was analyzed with Kaplan-Meier plotter. RESULTS: In the present study, the therapeutic effect of CTD on HCC was evaluated in vivo. We obtained the CTD-related transcriptional profiles, systematically and intuitively illustrated its possible pharmacological mechanisms in HCC through multiple targets and signalling pathways. These results revealed that the CTD-related differentially expressed genes were involved in autophagy, transcription factors (TFs) related transcriptional regulation, fatty acid metabolism and immune response in HCC. We found that MAPT, TOP2A, CENPF and MEFV were hub genes of CTD targets involved in autophagy regulation. Totally, 14 TFs have been confirmed to be critical for transcriptional regulation, and 33 TF targets were identified as the hub genes in transcriptional mis-regulation pathway in cancer. These TFs were associated with the immune response and immune cell infiltration. In addition, the downregulated genes were significantly enriched in metabolic regulation pathways, especially fatty acid metabolism after CTD treatment. Furthermore, the network of CTD associated miRNAs with these fatty acid metabolism-related targets was constructed in HCC. CONCLUSIONS: Taken together, our results comprehensively elucidated that CTD could act on multiple targets in HCC therapy, affecting autophagy, transcriptional regulation, the immune response and fatty acid metabolism. Our results provide a foundation for the study of the molecular mechanistic of CTD and its clinical application in the treatment of HCC.

Cantharidin suppresses hepatocellular carcinoma development by regulating EZH2/H3K27me3-dependent cell cycle progression and antitumour immune response.

BACKGROUND: Cantharidin (CTD) is a major ingredient of cantharis (Mylabris phalerata Pallas) and has been used extensively in traditional Chinese medicines. It has been shown to exhibit anticancer activity in multiple types of cancer, especially hepatocellular carcinoma (HCC). However, there is no systematic study on the relationships among the regulatory networks of its targets in HCC therapy. We focused on histone epigenetic regulation and the influence of CTD on the immune response in HCC. METHODS: We performed a comprehensive analysis of novel CTD targets in HCC based on network pharmacology and RNA-seq approaches. The mRNA levels of target genes were analyzed by qRT-PCR, and the corresponding protein levels were confirmed using enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining (IHC). ChIP-seq data were visualized by IGV software. The associations of gene transcript levels with the cancer immune score and infiltration level were investigated using TIMER. In vivo, the H22 mouse model of hepatocellular carcinoma was established by treatment with CTD and 5-Fu. The immune cell proportions in the blood were elevated in model mice, as shown by flow cytometry. RESULTS: We identified 58 targets of CTD, which were involved in various pathways in cancer, including apoptosis, the cell cycle, EMT and immune pathways. Moreover, we found that 100 EMT-related genes were differentially expressed after CTD treatment in HCC cells. Interestingly, our results confirmed that the EZH2/H3K27me3 -related cell cycle pathway is a therapeutic target of CTD in antitumour. In addition, we evaluated the influence of CTD on the immune response. Our data showed that the significantly enriched gene sets were positively correlated with the chemokine biosynthetic and chemokine metabolic modules. The proportions of CD4+/CD8 + T cells and B cells were increased, but the proportion of Tregs was decreased after treatment with CTD in vivo. Moreover, we found that the expression of the inflammatory factor and immune checkpoint genes PD-1/PD-L1 was significantly reduced in the mouse model. CONCLUSION: We performed a novel integrated analysis of the potential role of CTD in HCC treatment. Our results provide innovative insight into the mechanism by which cantharidin exerts antitumour effects by regulating target genes expression to mediate apoptosis, EMT, cell cycle progression and the immune response in HCC. Based on the effect of CTD on the immune response, it can be used as a potential effective drug to activate antitumour immunity for the treatment of liver cancer.

[Kinetics of adsorption of Pb2+ onto small river sediment].

The batch experiments of adsorption of Pb2+ onto small river sediments were conducted. The kinetics of the sorption process was analyzed. The results showed that the equilibrium time of adsorption increased with the increasing of sediment mass in solution, while both adsorbed Pb2+ on per unit of sediment and Pb2+ concentration in the solution after equilibrium decreased. More than 95% of Pb2+ in solution was removed when sediment contents larger than 0.6 g x L(-1). Both pseudo-first-order and pseudo-second-order kinetics were tested and it was found that the latter gave a better explanation of the adsorption process. The equilibrium adsorption capacities calculated from the pseudo-second-order model could represent the true value. There was no significant correlation between initial adsorption rate of Pb2+ and the amount of sediment in solution. However, the pseudo-second-order rate constant increased in the solution with more adsorbent, namely chemical adsorption controlled the process. Elovich equation could explain the mechanism of sorption in the solution with higher contents of sediment; nevertheless, the process of low concentration of adsorbent adsorbing Pb2+ disagreed well with Elovich equation. In terms of adsorption rate in the sorption, intra-particle diffusion dominated in the more sediment solution. On the other hand, multi-linearity was presented for the adsorption rate in less adsorbent solution. The first, sharper portion represented adsorption on the external surface. The second portion indicated Pb2+ diffused gradually into the interior of particles and intra-particle diffusion controlled.

[Heavy metals in environmental media around drinking water conservation area of Shanghai].

The levels of heavy metals in Shanghai drinking water conservation area were determined, and the spatial distributions and main sources of heavy metals were investigated. Moreover, the ecological risk assessment of heavy metals was conducted. Some conclusions can be drawn as follows: (1) The average concentrations of Cd, Hg, Pb, Cu, Zn, Ni, Cr and As in road dust were 0.80, 0.23, 148.45, 127.52, 380.57, 63.17, 250.38 and 10.37 mg x kg(-1) respectively. In terms of the pollution level, the values of soils were relatively lower, with the mean contents of 0.16 (Cd), 0.33 (Hg), 30.14 (Pb), 30.66 (Cu), 103.79 (Zn), 24.04 (Ni), 65.75 (Cr) and 6.31 mg x kg(-1) (As) severally; meanwhile the average levels of heavy metals in vegetables were 0.010 (Cd), 0.016 (Hg), 0.36 (Pb), 12.80 (Cu), 61.69 (Zn), 2.04 (Ni), 2.41 (Cr) and 0.039 mg x kg(-1) (As) respectively. (2) Semivariogram and multivariate analysis indicated that heavy metals pollution of soils was induced by anthropogenic activities mostly, and the pollutants produced by traffic were the major source of heavy metals in road dust. (3) The order for heavy metal enrichment coefficients of vegetables was as following: Zn (0.589) > Cu (0.412) > 0.102 (Ni) > Cd (0.059) > Cr (0.061) > Hg (0.056) > Pb (0.012) > As (0.007), and the results indicated that Cd and Zn in vegetables were mainly from the soils, and the other metals were probably from the pollutants in the atmosphere. (4) Sediments in drinking water conservation area were probably derived from soils around; however, there was no significant relationship between heavy metals contents of them. (5) The results of ecological risk assessment of heavy metals showed that heavy metals in soils were in no-warning to warning situation, and warning to light-warning situation for road dust and vegetables. The fuzzy synthesis judgment for all the environmental media around drinking water conservation area was warning to light-warning.

[Characteristics of heavy metal pollution in soil and dust of urban parks in Shanghai].

The contents of heavy metals in soils and dust of urban parks in Shanghai were studied, and the spatial distribution of heavy metals was also analyzed. The results showed that the average contents of Pb (55.06 mg x kg(-1)), Zn(198.54 mg x kg(-1)), Cu(44.57 mg x kg(-1)), Cr (77.01 mg x kg(-1)), Cd (0.40 mg x kg(-1)) and Ni (31.17 mg x kg(-1)) in soils were lower than them in the dust, which were 416.63 mg x kg(-1), 906.29 mg x kg(-1), 235.89 mg x kg(-1), 162.59 mg x kg(-1), 1.58 mg x kg(-1) and 92.19 mg x kg(-1) respectively. The heavy metals in soils except Ni and all the six heavy metals in dust were higher than the background values in Shanghai more or less. The heavy metals average contents in dust in the parks of city central area were lower than them in the parks of city surrounding area, and the heavy metals in soils have similar spatial distribution patterns except for Zn. However, in different districts, the distribution of heavy metals in the soils and dust were not regular. Pearson correlative analysis and principal component analysis indicated that the heavy meals accumulation of the soils and dust were induced by anthropogenic input, furthermore traffic and industry were the main pollution sources.