ABSTRACT
Mallotus japonicus is a valuable traditional medicinal plant in East Asia for applications as a gastrointestinal drug. However, the molecular components involved in the biosynthesis of bioactive metabolites have not yet been explored, primarily due to a lack of omics resources. In this study, we established metabolome and transcriptome resources for M. japonicus to capture the diverse metabolite constituents and active transcripts involved in its biosynthesis and regulation. A combination of untargeted metabolite profiling with data-dependent metabolite fragmentation and metabolite annotation through manual curation and feature-based molecular networking established an overall metabospace of M. japonicus represented by 2129 metabolite features. M. japonicus de novo transcriptome assembly showed 96.9% transcriptome completeness, representing 226,250 active transcripts across seven tissues. We identified specialized metabolites biosynthesis in a tissue-specific manner, with a strong correlation between transcripts expression and metabolite accumulations in M. japonicus. The correlation- and network-based integration of metabolome and transcriptome datasets identified candidate genes involved in the biosynthesis of key specialized metabolites of M. japonicus. We further used phylogenetic analysis to identify 13 C-glycosyltransferases and 11 methyltransferases coding candidate genes involved in the biosynthesis of medicinally important bergenin. This study provides comprehensive, high-quality multi-omics resources to further investigate biological properties of specialized metabolites biosynthesis in M. japonicus.
Subject(s)
Gene Expression Regulation, Plant , Gene Regulatory Networks , Mallotus Plant/metabolism , Metabolome , Plant Proteins/metabolism , Transcriptome , Gene Expression Profiling , Mallotus Plant/genetics , Mallotus Plant/growth & development , Organ Specificity , Phylogeny , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Roots/genetics , Plant Roots/growth & development , Plant Roots/metabolismABSTRACT
Anoctamin1 (ANO1), a calcium-activated chloride channel, is frequently overexpressed in several cancers, including human prostate cancer and oral squamous cell carcinomas. ANO1 plays a critical role in tumor growth and maintenance of these cancers. In this study, we have isolated two new compounds (1 and 2) and four known compounds (3-6) from Mallotus apelta. These compounds were evaluated for their inhibitory effects on ANO1 channel activity and their cytotoxic effects on PC-3 prostate cancer cells. Interestingly, compounds 1 and 2 significantly reduced both ANO1 channel activity and cell viability. Electrophysiological study revealed that compound 2 (Ani-D2) is a potent and selective ANO1 inhibitor, with an IC50 value of 2.64 µM. Ani-D2 had minimal effect on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activity and intracellular calcium signaling. Notably, Ani-D2 significantly reduced ANO1 protein expression levels and cell viability in an ANO1-dependent manner in PC-3 and oral squamous cell carcinoma CAL-27 cells. In addition, Ani-D2 strongly reduced cell migration and induced activation of caspase-3 and cleavage of PARP in PC-3 and CAL-27 cells. This study revealed that a novel ANO1 inhibitor, Ani-D2, has therapeutic potential for the treatment of several cancers that overexpress ANO1, such as prostate cancer and oral squamous cell carcinoma.
Subject(s)
Anoctamin-1/antagonists & inhibitors , Mallotus Plant/metabolism , Plant Extracts/pharmacology , Animals , Anoctamin-1/metabolism , Anoctamin-1/physiology , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Calcium/metabolism , Calcium Signaling/drug effects , Carcinoma, Squamous Cell/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Chloride Channels/metabolism , Humans , Mouth Neoplasms/metabolism , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/metabolism , Neoplasm Proteins/physiology , PC-3 Cells , RatsABSTRACT
Five prenylflavonoids, 6-prenylnaringenin (1), 8-prenylnaringenin (2), 7-O-methyl-8-prenylnaringenin (3), 7-O-methyl-6-prenylnaringenin (4), and 4'-O-methyl-6-prenylnaringenin (5), were isolated from the traditional herb Mallotus conspurcatus Croizat (Euphorbiaceae). Compounds 1-5 revealed cytotoxic activity against cervical cancer (HeLa) cells with IC50 values ranging from 10.08 to 60.16â µm by MTT method, and interestingly, these prenylflavonoids were less toxic to normal HL-7702 cells. Furthermore, compounds 1 and 5 could inhibit the c-myc expression and telomerase activity and cause mitochondrial dysfunction. These findings might contribute to a better understanding of the biological activities of prenylflavonoids and lay the foundation for further studies on the cytotoxic activity of natural products isolated from M. conspurcatus.
Subject(s)
Flavonoids/chemistry , Mallotus Plant/chemistry , Apoptosis/drug effects , Caspase 3/metabolism , Cell Line , Down-Regulation , Flavanones/chemistry , Flavanones/isolation & purification , Flavanones/pharmacology , Flavonoids/isolation & purification , Flavonoids/pharmacology , HeLa Cells , Humans , Mallotus Plant/metabolism , Membrane Potential, Mitochondrial/drug effects , Mitochondria/drug effects , Mitochondria/metabolism , Plant Extracts/chemistry , Plant Roots/chemistry , Plant Roots/metabolism , Proto-Oncogene Proteins c-myc/metabolism , Reactive Oxygen Species/metabolism , Telomerase/metabolismABSTRACT
Some Mallotus species are used in traditional medicine in Vietnam. To use certain species in Western medicines or as food supplements, they should be identified and quality control should be more strict, for instance, to avoid the erroneous switching of species. In species with interesting activities, the compounds responsible for them should be identified. For these identifications, HPLC fingerprint methodology can be used. In this paper, HPLC fingerprints of different lengths were developed for a number of Mallotus species. Secondly, a multivariate regression model was constructed to model the antioxidant activity of the Mallotus samples from the HPLC fingerprints with the aim to indicate peaks possibly responsible for this activity. For this purpose, after data pretreatment, the calibration technique partial least squares (PLS) was applied.