ABSTRACT
BACKGROUND: Pine needle oil from crude extract of pine needles has anti-tumor effects, but the effective component is not known. METHODS: In the present study, compounds from a steam distillation extract of pine needles were isolated and characterized. Alpha-pinene was identified as an active anti-proliferative compound on hepatoma carcinoma BEL-7402 cells using the MTT assay. RESULTS: Further experiments showed that α-pinene inhibited BEL-7402 cells by arresting cell growth in the G2/M phase of the cell cycle, downregulating Cdc25C mRNA and protein expression, and reducing cycle dependence on kinase 1(CDK1) activity. CONCLUSION: Taken together, these findings indicate that α-pinene may be useful as a potential anti-tumor drug.
Subject(s)
Carcinoma, Hepatocellular/drug therapy , Cell Proliferation/drug effects , Liver Neoplasms/drug therapy , M Phase Cell Cycle Checkpoints/drug effects , Monoterpenes/pharmacology , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Bicyclic Monoterpenes , CDC2 Protein Kinase , Cell Line, Tumor , Cyclin-Dependent Kinases/biosynthesis , Down-Regulation , Humans , Pinus/metabolism , Plant Extracts/pharmacology , RNA, Messenger/biosynthesis , cdc25 Phosphatases/biosynthesis , cdc25 Phosphatases/geneticsABSTRACT
OBJECTIVE: To analyze the chemical composition of essential oil from Thymus citriodorus and its toxic effect on liver cancer cells. METHODS: The essential oil from Thymus citriodorus leaves was extracted by steam distillation, and GC-MS was used for analyzing chemical composition. 35 components were identified which accounted for 95.44% of the total peak area. The main components were borneol (28.82%), thymol (14.43%), 3, 7-dimethyl-1, 6-octadiene-3-ol (8.26%), 1-methyl-4-[alpha-hydroxy-isopropyl] cyclohexene (8.23%) and terpenes camphor (5.1%), et. al. The toxic effects on HepG2 cells and expression of NF-kappabeta65 were measured by MTT assay and confocal laser scanning microscopy, respectively. RESULTS: The IC50 of HepG2 cells inhibition was 0.34%. The mean fluorescence intensity of NF-kappabeta65 expression was as follows: control group 323.25, 2(-10) concentration group 84.18, 2(-11) concentration group 197.93 and 2(-12) concentration group 261.43. CONCLUSION: The essential oil from Thymus citriodorus leaves has strong toxic effects. The induced apoptosis mechanism may be associated with the expression of NF-kappabeta65.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Cell Proliferation/drug effects , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Thymus Plant/chemistry , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Camphanes/analysis , Drug Screening Assays, Antitumor , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Drugs, Chinese Herbal/pharmacology , Gas Chromatography-Mass Spectrometry , Hep G2 Cells , Humans , Inhibitory Concentration 50 , Liver Neoplasms/pathology , NF-kappa B/metabolism , Oils, Volatile/isolation & purification , Plant Leaves/chemistry , Thymol/analysisABSTRACT
Radix Lithosperm eyrthrorhizon is a common prescription compound in traditional Chinese medicine. Shikonin is a major component of Radix Lithospermi and has various biological activities. We have investigated the inhibitory effect of shikonin on the growth of adenovirus type 3 (AdV3) in vitro. The antiviral function of shikonin against AdV3 and its virus inhibition ratio were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method (MTT). The expression of hexon protein in AdV3 was determined by immunofluorescence assay using laser scanning confocal microscopy (LSCM) and Western blot analysis. In addition, the rate of apoptosis in cells infected by AdV3 was determined by flow cytometry. Shikonin (0.0156-1 µM) inhibited growth of AdV3 in a concentration-dependent manner with a virus inhibition rate of 23.8-69.1%. Expression of hexon protein in AdV3 was higher in the virus control group than in the shikonin-treated groups as determined by immunofluorescence assay and Western blotting (p<0.05). The rate of shikonin-treated HeLa cell apoptosis had a statistically significant decrease with increasing concentration of drug (p<0.05). Our data demonstrate that shikonin possesses anti-AdV3 capabilities and that the potential antiviral mechanism might involve inhibiting the degree of apoptosis and hexon protein expression of AdV.
Subject(s)
Adenoviridae Infections/drug therapy , Adenoviridae/drug effects , Antiviral Agents/pharmacology , Capsid Proteins/metabolism , Drugs, Chinese Herbal/pharmacology , Lithospermum/chemistry , Naphthoquinones/pharmacology , Adenoviridae/growth & development , Antiviral Agents/therapeutic use , Apoptosis/drug effects , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/therapeutic use , HeLa Cells , Humans , Naphthoquinones/therapeutic use , Phytotherapy , Plant RootsABSTRACT
OBJECTIVE: To study the effects of essential oil extracted from pine needles on HepG2 cell line. METHODS: HepG2 cells were treated with essential oil extracted from pine needles. Cell growth rate was determined with MTF assay, cell morphologic changes were examined under transmission electromicroscope and HE straining. Flow cytometry was used to exmine apoptotic cells. Bcl-2 gene expression was determined by flow cytometry and telomerase activity by TRAP assay. RESULTS: Essential oils from pine needles could not only repress the growth of HepG2 cells significantly, but also induce apoptosis to them. Both dose-effect and time-effect relationship could be confirmed. Typical morphology changes of apoptosis such as nuclear enrichment and karyorrhexis were observed through transmission electromicroscope and HE straining. Telomerase activity was down regulated in the essential oil extracted from pine needles induced apoptotic cells. The expression of bcl-2 gene was suppressed after the essential oil from pine needles treatement. CONCLUSION: The essential oil extracted from pine needles can inhibit cell growth of HepG2 cell line and induce apoptosis, which may associate with inhibition of telomerase activity and bcl-2 may be involved in the regulation of telomerase activity.