ABSTRACT
Antrodia cinnamomea is extensively used as a traditional medicine to prevention and treatment of liver cancer. However, its comprehensive chemical fingerprint is uncertain, and the mechanisms, especially the potential therapeutic target for anti-hepatocellular carcinoma (HCC) are still unclear. Using UPLC‒Q-TOF/MS, 139 chemical components were identified in A. cinnamomea dropping pills (ACDPs). Based on these chemical components, network pharmacology demonstrated that the targets of active components were significantly enriched in the pathways in cancer, which were closely related with cell proliferation regulation. Next, HCC data was downloaded from Gene Expression Omnibus database (GEO). The Cancer Genome Atlas (TCGA) and DisGeNET were analyzed by bioinformatics, and 79 biomarkers were obtained. Furtherly, nine targets of ACDP active components were revealed, and they were significantly enriched in PI3K/AKT and cell cycle signaling pathways. The affinity between these targets and their corresponding active ingredients was predicted by molecular docking. Finally, in vivo and in vitro experiments showed that ACDPs could reduce the activity of PI3K/AKT signaling pathway and downregulate the expression of cell cycle-related proteins, contributing to the decreased growth of liver cancer. Altogether, PI3K/AKT-cell cycle appears as the significant central node in anti-liver cancer of A. Cinnamomea.
ABSTRACT
Objective: To excavate the terpenoid synthesis and metabolism-related gene function and screen the interaction protein and fingerprint analysis of Antrodia cinnamomea mycelium, a cDNA library from A. cinnamomea mycelia was constructed and the EST sequences were analyzed. Methods: The cDNA library from the A. cinnamomea mycelium was constructed by the Gateway technique. A part of EST sequences about the bioinformatics, functional annotation and EST-SSR were analyzed. Results: The cDNA library of the A. cinnamomea mycelium was constructed successfully. The recombinant rate of the cDNA library was 95%, the titer of the library was 6.1 × 106 cfu/mL, the total cloning number was 1.2 × 107 cfu, the length of cDNA was between 300-2 000 bp with an average length of 1 000 bp. The clones were randomly sequenced and 65 valid ESTs were obtained. After being compared in the Genbank database, 45 ESTs had a definite annotation, and 18 ESTs were unnamed and hypothetical protein. The results with GO functional annotation showed that the ESTs involved the cell composition, transport, catalytic activity, regulation functions and etc. It contained 271 SSRs of all the ESTs in total. The nucleotide repeats in A. cinnamomea were abundant, among which dinucleotide and trinucleotide repeat units were more common accounting for 94.23%. Conclusion: The cDNA library from the A. cinnamomea mycelium and its ESTs related biological information were preliminarily identified, which will provide a theoretical foundation for research the mycelium genomics of A. cinnamomea.
ABSTRACT
Head and neck squamous cell carcinoma (HNSCC) is one of the most common malignant tumors. Ethanol extract of Antrodia cinnamomea (EEA) has been widely studied for its health benefits including anticancer effects. The purpose of this study was to assess the effects of EEA on HNSCC. Cell proliferation, transwell, and wound healing assays were performed. The impact of EEA on tumor growth was investigated using a xenograft model. Expressions of migration-related proteins (MMP-2, MMP-9, TIMP-1, and TIMP-2) and apoptosis-related proteins (cleaved caspase-9 and cleaved PARP) were determined using western blot analysis. The results indicated that EEA significantly inhibited the capacities of proliferation, invasion, and migration of HNSCC cells in a dose-dependent manner. Cleaved caspase-9 and cleaved PARP expressions were increased in cells treated with an increasing concentration of EEA, which suggested that EEA induced apoptosis of HNSCC. MMP-2 and MMP-9 were downregulated when cells were administered EEA, while TIMP-1 and TIMP-2 were not affected, which uncovered the mechanisms mediating the EEA-induced inhibition on cell invasion and migration. The animal experiment also suggested that EEA inhibited tumor growth. Our study confirmed the inhibitive effects of EEA on cell proliferation, invasion, and migration of HNSCC in vitro and in vivo, providing the basis for further study of the application of EEA as an effective candidate for cancer treatment.
Subject(s)
Humans , Animals , Female , Rabbits , Biological Products/pharmacology , Ethanol/pharmacology , Antrodia/chemistry , Squamous Cell Carcinoma of Head and Neck/pathology , Lung Neoplasms/pathology , Time Factors , Blotting, Western , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Ethanol/isolation & purification , Squamous Cell Carcinoma of Head and Neck/drug therapy , Lung Neoplasms/drug therapy , Mice, Inbred BALB CABSTRACT
Objective: To investigate the effects of phytohormones and lignin on the growth of Antrodia cinnamomea mycelium, and to determine the content of crude polysaccharide and crude triterpenoid and antioxidant activity of cultured products. Methods The product cultured by solid-state fermentation in petri dish was ultrasonically extracted. The phenol-sulfuric acid method was selected to determine the content of crude polysaccharide of the extract, and the vanillin-glacial acetic acid method was used to determine the content of crude triterpenoids of the extract. The half-clearing concentrations (IC50) of DPPH free radicals and ABTS free radicals were indexes for evaluating the anti-oxidant activity of the culture product. Results Sampling near the outer edge of the mycelium layer after 20 d of culture was performed by the activation method to obtain inoculated raw materials with better growth activity; The basal medium with good growth, high content of crude polysaccharide and crude triterpenoids was modified PCA medium; On the basis of this medium, the mycelia with 0.5 g/L lignin added into the medium grew fastest, and the diameter of mycelium layer increased to 1.19 times of the control group; When the concentration of IBA was 0.5 mg/L, the dry weight of mycelium was increased by 89.51% compared with the control group, and the yield of crude polysaccharide was increased by 130.57% compared with the control group, which was much higher than other groups. The content and yield of triterpenoids were also increased significantly, 61.31% higher than the control group, and crude triterpenoids yield reached 133.24 mg/L; The content of crude triterpenoids in mycelium was the highest (5.62%) when adding 0.5 g/L powder of Cinnamomum kanehirai, which was 84.26% higher than that of the control group; In addition, the addition of plant hormones and lignin to the culture medium has a certain effect on the anti-oxidant capacity of the Antrodia cinnamomea mycelium, on the whole, the experimental group had good anti-oxidant activity after adding different substances. Conclusion By adding phytohormone and lignin to the modified PCA medium, the growth of the mycelium of Antrodia cinnamomea can be effectively promoted, the content of the active ingredient can be increased, and the anti-oxidant activity can be enhanced.
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Objective To clone the squalene epoxidase gene of Antrodia cinnamomea (AcSE) and analyze the bioinformatics and expression of the gene. Methods AcSE was cloned by rapid-amplification of cDNA ends (RACE) from cDNA of A. cinnamomea. The physical and chemical properties of AcSE protein were analyzed, and its secondary structure, tertiary structure, and function were predicted by using bioinformatics analysis. The expression of AcSE in mycelium and fruit body of A. cinnamomea at different culture time was detected by using quantitative real-time PCR (qRT-PCR). Results The full-length cDNA sequence of AcSE were 1 446 bp (Genbank: KT070558), encoding a 481-amino-acid polypeptide. The molecular weight of AcSE was 53 300 and pI was 6.36. Domain analysis results showed that AcSE had three transmembrane domains without coiled-coil structure, and hydrophobic and hydrophilic regions existed alternately. The gDNA sequence of AcSE was 1 607 bp, contained four exons and three introns. A gene expression analysis by relative qRT-PCR showed that the highest expression level of AcSE was in mycelia incubated for 7 d of A. cinnamomea, and it was 7.89 times than that in fruiting body, and a gradual decline was observed with the extension of the culture time. Conclusion Gene AcSE was firstly cloned from A. cinnamomea, and it would lay a foundation for exploring the mechanism of terpenoid biosynthesis in A. cinnamomea.
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Objective To compare and study species homology of two kinds of Antrodia Cinnamomea; To conduct the study on quality control. Methods HPLC fingerprint of two kinds of Antrodia Cinnamomea was applied. The HPLC fingerprints were determined on an Diamonsil C18 (2) column (250 mm × 4.6 mm, 5 μm) eluted with the mobile phase consisting of 1% formic acid solution and acetonitrile in gradient mode at a flow rate of 0.8 mL/min; the detection wavelength was set at 254 nm; the column temperature was 30 ℃. Results There were 9 common peaks in the HPLC fingerprints of Antrodia Cinnamomea. The similarities varied from 0.906 to 0.995 and from 0.956 to 0.998 in 11 batches and 10 batches of two kinds of Antrodia Cinnamomea, respectively. Conclusion The method is simple and with good reproducibility, which shows that the HPLC fingerprint and infrared spectrum share similarity for two kindsof Antrodia Cinnamomea.
ABSTRACT
Antrodia cinnamomea is a medicinal fungi originated in Taiwan. It is rich of triterpene substances, polysaccharide, and adenosine, and exhibits pharmacological activity of antitumor, immunomodulatory, anti-inflammatory effects, and so on. In recent years, it gradually becomes the hot spot about application and research of A. cinnamomea due to more and more research reports. This paper reviews the biological characteristics, chemical constituents, and pharmacological effects of A. cinnamomea, which can provide the references for future research and application of A. cinnamomea.
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Extensive in vitro studies reveal that multiple intracellular targets of Antrodia cinnomomea affect cell growth,apoptosis,angiogenesis,invasion and metastasis.These intracellular targets include tumor suppressor gene,cell cycle regulatory protein,transcription factor,angiogenic and metastatic factors,and apop-totic and survival regulators.In addition,Antrodia cinnomomea has immunomodulatory propertie,which plays an anti-cancer effect indirectly by means of enhancing immunity.
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Objective: To explore whether Antrodia cinnamomea can prolong the survival time of stroke-prone spontaneously hypertensive rats, and to investigate the underlying mechanisms from the proteomics perspective. Methods: A total of 80 male stroke-prone spontaneously hypertensive rats were randomly divided into two groups (control group and Antrodia cinnamomea treated group, n= 40). The animals were intragastrically given Antrodia cinnamomea (150 mg/kg). Then the death of the animals in the two groups was observed. Another 6 stroke-prone spontaneously hypertensive rats were randomly divided into drug treatment group and control group, with the drug treatment group intragastrically given Antrodia cinnamomea (150 mg/kg), and the brain proteomics were compared between the two groups 90 days later, with WKY rats used as normal controls. The differentially expressed proteins before and after drug treatment were subjected to mass spectrometry analysis, and the results of mass spectrometry analysis were further confirmed by Western blotting analysis. Results: Antrodia cinnamomea significantly prolonged the survival time of stroke-prone spontaneously hypertensive rats (P < 0.05), and proteomics results showed that Antrodia cinnamomea increased the expression of glutathione S-transferase (GST) and superoxide dismutase (SOD), which were also confirmed by Western blotting analysis. Further study revealed that the total anti-oxidative capability (T-AOC) of the animals was increased (T-AOC; [66.48 ± 16.17] U/g vs [124.75 ± 28.43] U/g, P< 0.05), which was manifested by the increased activity of GST and SOD (GST: [40.33 ± 5.24] U/mg vs [70.50 ± 6.24] U/ mg, P<0.05; SOD; [109.25 ± 23.61] U/mg vs [192.60 ± 23.95] U/mg, P<0.05), and decreased levei of malondialdehyde ([3.96 ± 0.45] nmol/mg vs [2.04 ± 0.31] nmol/mg, P<0.05). Conclusion: Long-term treatment with Antrodia cinnamomea can prolong the lifespan of stroke-prone spontaneously hypertensive rats, which might be related to the increased activity of anti-oxidative enzymes and the decreased oxidative damage.