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1.
Exp Ther Med ; 14(1): 59-64, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28672893

ABSTRACT

Rice is a staple food in numerous countries around the world. Anthocyanins found in black rice have been reported to reduce the risk of certain diseases, but the effects of crude extract of anthocyanins from Asia University-selected purple glutinous indica rice (AUPGA) on immune responses have not yet been demonstrated. The current study aimed to investigate whether AUPGA treatment could affect immune responses in murine leukemia cells in vivo. Murine acute myelomonocytic leukemia WEHI-3 cells were intraperitoneally injected into normal BALB/c mice to generate leukemia mice. A total of 50 mice were randomly divided into five groups (n=10 in each group) and were fed a diet supplemented with AUPGA at 0, 20, 50 or 100 mg/kg for three weeks. All mice were weighed and the blood, liver and spleen were collected for further experiments. The results indicated that AUPGA did not significantly affect animal body weight, but significantly increased spleen weight (P<0.05) and decreased liver weight (P<0.05) when compared with the control group. AUPGA significantly increased the T cell (CD3) population at treatments of 20 and 100 mg/kg (P<0.05). However, it only significantly increased the B cell (CD19) population at a treatment of 20 mg/kg (P<0.05). Furthermore, AUPGA at 50 and 100 mg/kg significantly increased the monocyte (CD11b) population and the level of macrophages (Mac-3; P<0.05 for both). AUPGA at 50 and 100 mg/kg significantly promoted macrophage phagocytosis in peripheral blood mononuclear cells (P<0.05), and all doses of AUPGA treatment significantly promoted macrophage phagocytotic activity in the peritoneum (P<0.05). AUPGA treatment significantly decreased natural killer cell activity from splenocytes (P<0.05). Finally, AUPGA treatment at 20 mg/kg treatment significantly promoted T cell proliferation (P<0.05), and treatment at 50 and 100 mg/kg significantly decreased B cell proliferation compared with the control group (P<0.05).

2.
Am J Chin Med ; 44(3): 637-61, 2016.
Article in English | MEDLINE | ID: mdl-27109154

ABSTRACT

Casticin, a polymethoxyflavone occurring in natural plants, has been shown to have anticancer activities. In the present study, we aims to investigate the anti-skin cancer activity of casticin on melanoma cells in vitro and the antitumor effect of casticin on human melanoma xenografts in nu/nu mice in vivo. A flow cytometric assay was performed to detect expression of viable cells, cell cycles, reactive oxygen species production, levels of [Formula: see text] and caspase activity. A Western blotting assay and confocal laser microscope examination were performed to detect expression of protein levels. In the in vitro studies, we found that casticin induced morphological cell changes and DNA condensation and damage, decreased the total viable cells, and induced G2/M phase arrest. Casticin promoted reactive oxygen species (ROS) production, decreased the level of [Formula: see text], and promoted caspase-3 activities in A375.S2 cells. The induced G2/M phase arrest indicated by the Western blotting assay showed that casticin promoted the expression of p53, p21 and CHK-1 proteins and inhibited the protein levels of Cdc25c, CDK-1, Cyclin A and B. The casticin-induced apoptosis indicated that casticin promoted pro-apoptotic proteins but inhibited anti-apoptotic proteins. These findings also were confirmed by the fact that casticin promoted the release of AIF and Endo G from mitochondria to cytosol. An electrophoretic mobility shift assay (EMSA) assay showed that casticin inhibited the NF-[Formula: see text]B binding DNA and that these effects were time-dependent. In the in vivo studies, results from immuno-deficient nu/nu mice bearing the A375.S2 tumor xenograft indicated that casticin significantly suppressed tumor growth based on tumor size and weight decreases. Early G2/M arrest and mitochondria-dependent signaling contributed to the apoptotic A375.S2 cell demise induced by casticin. In in vivo experiments, A375.S2 also efficaciously suppressed tumor volume in a xenotransplantation model. Therefore, casticin might be a potential therapeutic agent for the treatment of skin cancer in the future.


Subject(s)
Antineoplastic Agents, Phytogenic , Apoptosis/drug effects , Apoptosis/genetics , Flavonoids/pharmacology , Flavonoids/therapeutic use , Melanoma/drug therapy , Melanoma/pathology , Mitochondria , NF-kappa B , Phytotherapy , Signal Transduction , Skin Neoplasms/drug therapy , Skin Neoplasms/pathology , Animals , Caspases/metabolism , Disease Models, Animal , G2 Phase Cell Cycle Checkpoints/drug effects , G2 Phase Cell Cycle Checkpoints/genetics , Heterografts , Humans , Melanoma/genetics , Melanoma/metabolism , Mice , Mice, Nude , Neoplasm Transplantation , Neoplastic Cells, Circulating , Reactive Oxygen Species/metabolism , Skin Neoplasms/genetics , Skin Neoplasms/metabolism
3.
Environ Toxicol ; 31(8): 957-69, 2016 Aug.
Article in English | MEDLINE | ID: mdl-25689151

ABSTRACT

Cancer is the second cause of death in children. Osteosarcoma is the most common primary malignancy of solid bone cancer primarily affecting adolescents and young adults. In the Chinese population, the crude extract of Rheum palmatum L. (CERP) has been used for treating different diseases, including SARS, rheumatoid arthritis, coxsackievirus B3, and human colon cancer cell, pancreatic cancer. There are no reports on CERP and human osteosarcoma cells. The present study examined effects of CERP on cytotoxicity including cell cycle distribution and cell death (apoptosis) in U-2 OS human osteosarcoma cells. CERP significantly induced S phase arrest in U-2 OS cells in a dose-dependent. CERP produced DNA damage and DNA condensation. Other effects of CERP were stimulation of ROS and Ca(2+) , mitochondria impairment, and activation of caspase-3, -8, and -9. CERP increased the levels of Bax, Bak, Bad, cyclin B, Fas, PARP, GRP78, GADD153, AIF, Endo G, Calpain-2, p21, and p27, but decreased the levels of Bcl-2, BCL-X, XIAP, Akt, CDC25A, CDK2, Cyclin A, and Cyclin E of U-2 OS cells. It was also observed that CERP promoted the expression of AIF, Endo G, GADD153, and cytochrome c. These results indicate that CERP has anticancer effects in vitro and provide the foundation for in vivo studies of animal models of osteosarcoma. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 957-969, 2016.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Drugs, Chinese Herbal/pharmacology , S Phase Cell Cycle Checkpoints/drug effects , Apoptosis Regulatory Proteins/metabolism , Bone Neoplasms/drug therapy , Bone Neoplasms/pathology , Calcium Signaling , Cell Line, Tumor , Cell Survival/drug effects , DNA Damage , Drug Screening Assays, Antitumor , Endoplasmic Reticulum Chaperone BiP , Humans , Membrane Potential, Mitochondrial/drug effects , Mitochondria/drug effects , Mitochondria/metabolism , Osteosarcoma/drug therapy , Osteosarcoma/pathology , Protein Transport , Reactive Oxygen Species/metabolism , Rheum/chemistry
4.
In Vivo ; 29(6): 711-28, 2015.
Article in English | MEDLINE | ID: mdl-26546528

ABSTRACT

BACKGROUND/AIM: Lung cancer is one of the most common malignancies and a predominant cause of cancer-related death. It can metastasize in almost all organs, and currently, while new cases are increasing, treatment is still insufficient. Bisdemethoxycurcumin (BDMC), one of the components of turmeric, has been known to possess biological activities. However, the effects of BDMC on the genetic level remain unclear. MATERIALS AND METHODS: Human lung cancer NCI-H460 cells were treated with 35 µM BDMC for 24 h and cells were harvested for total RNA extraction. The purified RNA was used for cDNA synthesis, labeling, microarray hybridization, and flour-labeled cDNA on-chip hybridization. The expression Console software (Affymetrix) with default RNA parameters was used to detect and quantitate concentrations of fluorescent molecules. The key genes involved and their possible interaction pathways were analyzed by the GeneGo software. RESULTS: Seven genes, such as CCNE2 (cyclin E), associated with cell cycle, were over 4-fold overexpressed, 22 genes, such as ERCC6L (excision repair cross-complementing rodent repair deficiency, complementation group 6-like) associated with DNA damage and repair, were from 3- to 4-fold overexpressed and 266, such as cell division cycle, S-phase associated kinase and associated with cell death, genes were from 2- to 3-fold overexpressed. CONCLUSION: BDMC induced changes in gene expression that may reveal cytotoxic information on the genetic level while presenting novel biomarkers or targets for treatment of human lung cancer in the future.


Subject(s)
Carcinoma, Non-Small-Cell Lung/drug therapy , Curcumin/analogs & derivatives , Gene Expression Regulation, Neoplastic/drug effects , Neoplasm Proteins/biosynthesis , Apoptosis/drug effects , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Cell Cycle/drug effects , Cell Cycle/genetics , Cell Line, Tumor , Cell Movement/drug effects , Cell Movement/genetics , Curcuma/chemistry , Curcumin/administration & dosage , DNA Damage/drug effects , Diarylheptanoids , Humans , Microarray Analysis , Neoplasm Invasiveness/genetics , Neoplasm Proteins/genetics , Signal Transduction/drug effects
5.
Am J Chin Med ; 43(7): 1453-69, 2015.
Article in English | MEDLINE | ID: mdl-26477797

ABSTRACT

Solanum lyratum (SLEC) Thunberg (Solanaceae) has been used as a traditional herbal medicine in China for centuries. Numerous studies have shown that SLEC Thunberg (Solanaceae) extract inhibited cancer cell growth in vitro. Herein, we investigated cell death-induced by EcoAc, water, chloroform, butanol extract of SLEC in human oral cancer cell lines (HSC-3, SAS, and CAL-27) in vitro. Different SLEC extract induced cytotoxic effects in human oral cancer cells were examined by contrast phase microscopy. We selected the chloroform extract of SLEC to examine the cytotoxic effects by using DAPI staining, comet assays, flow cytometric assay, Western blotting and examination of confocal laser microscopy. SLEC decreased the percentage of viable cells, induced G0/G1 arrest and apoptosis. These effects were concentration- and time-dependent manners. SLEC increased protein levels of p21, p16, CDK2, and cyclin D1 in HSC-3, SAS, and CAL-27 cells. Also, SLEC increased CDK6 in HSC-3 and CAL-27 cells, but inhibited CDK6 in SAS cells. Cyclin E in HSC-3 and SAS cells was increased by SLEC, but it was inhibited in CAL-27 cells. SLEC suppressed the anti-apoptotic proteins Bcl-2 and Bcl-xl, but increased the pro-apoptotic proteins Bax and Bad in HSC-3, SAS, and CAL-27 cells. SLEC promoted the production of reactive oxygen species (ROS) and Ca²âº, decreased the mitochondrial membrane potential (Δψm) and stimulated NO production in HSC-3, SAS, and CAL-27 cells. Specific caspase inhibitors (caspase-8 inhibitor: Z-IETD-FMK; caspase-9 inhibitor: Z-LEHD-FMK and caspase-3 inhibitor: Z-DEVD-FMK) for caspase-8, -9, and -3 blocked SLE-activated caspase-8, -9, and -3 activities which were associated with an increase in the percentage of viable cells. Taken together, SLE induced G0/G1 arrest and apoptosis via extrinsic- and intrinsic-dependent pathways in HSC-3, SAS, and CAL-27 cells.


Subject(s)
Antineoplastic Agents, Phytogenic , Apoptosis/drug effects , Carcinoma, Squamous Cell/pathology , Caspases/metabolism , Chloroform , Cyclin-Dependent Kinase Inhibitor p21/metabolism , G1 Phase Cell Cycle Checkpoints/drug effects , Mitochondria/metabolism , Mouth Neoplasms/pathology , Neoplasm Proteins/metabolism , Plant Extracts/pharmacology , Reactive Oxygen Species/metabolism , Resting Phase, Cell Cycle/drug effects , Signal Transduction/drug effects , Solanum/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Calcium/metabolism , Carcinoma, Squamous Cell/metabolism , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p16 , G1 Phase Cell Cycle Checkpoints/genetics , Humans , Membrane Potential, Mitochondrial/drug effects , Mouth Neoplasms/metabolism , Plant Extracts/isolation & purification , Resting Phase, Cell Cycle/genetics
6.
In Vivo ; 29(4): 453-60, 2015.
Article in English | MEDLINE | ID: mdl-26130790

ABSTRACT

Numerous clinical anticancer drugs are obtained from natural plants and Hedyotis diffusa Willd (EEHDW) has been used as a major component in Traditional Chinese medicine formulas since a long time. Ethanol extracts of EEHDW have been shown to possess various biological activities including anticancer function in vitro. Our earlier studies have shown that EEHDW affects immune responses in WEHI-3-generated leukemia mice, but EEHDW has not been reported to affect immune responses in a normal mouse model. Herein, we investigated whether EEHDW could affect immune responses on normal murine cells in vivo. Normal BALB/c mice were orally treated with or without EEHDW at 0, 16, 32, and 64 mg/kg or 32 mg/kg by i.p. for 3 weeks, then were weighed, and blood, liver and spleen samples were collected for further experiments. Results indicated that EEHDW did not significantly affect body and liver weight but significantly increased the spleen weight by i.p. treatment when compared to control groups. Flow cytometric assays indicated that EEHDW promoted CD11b levels at 16, 32 and 64 mg/kg oral treatment, CD19 levels at 16, 32, 64 mg/kg oral treatment and i.p. treatment, and Mac-3 levels at 16, 32 and 64 mg/kg oral treatment, however, it did not significantly affect the levels of CD3. Oral treatment with 16 and 32 mg/kg of EEHDW significantly decreased macrophage phagocytosis from PBMC; 32 mg/kg of EEHDW by i.p. treatment significantly increased phagocytosis activity of macrophages obtain from the peritoneal cavity. EEHDW at 32 mg/kg by i.p. treatment led to an increase of NK cell activities compared to oil control groups. EEHDW at 32 mg/kg of EEHDW by i.p. treatment increased B- and T-cell proliferation. Based on these observations, EEHDW seems to have promoted immune responses in this murine model.


Subject(s)
Hedyotis/chemistry , Immune System/drug effects , Immunity/drug effects , Immunologic Factors/pharmacology , Plant Extracts/pharmacology , Animals , Antigens, Surface/metabolism , B-Lymphocytes/drug effects , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , Biomarkers , Blood Cells/drug effects , Blood Cells/metabolism , Body Weight/drug effects , Cytotoxicity, Immunologic/drug effects , Immunologic Factors/administration & dosage , Killer Cells, Natural/drug effects , Killer Cells, Natural/immunology , Killer Cells, Natural/metabolism , Lymphocyte Activation/drug effects , Lymphocyte Activation/immunology , Macrophages/drug effects , Macrophages/immunology , Macrophages/metabolism , Male , Mice , Mice, Inbred BALB C , Organ Size/drug effects , Phagocytosis/drug effects , Phagocytosis/immunology , Plant Extracts/administration & dosage , Plant Extracts/chemistry , T-Lymphocytes/drug effects , T-Lymphocytes/immunology , T-Lymphocytes/metabolism
7.
Anticancer Res ; 35(4): 2077-84, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25862863

ABSTRACT

Gallic acid (GA), a phenolic compound naturally present in plants, used as an antioxidant additive in food and in the pharmaceutical industry, may have cancer chemopreventive properties. In the present study, we investigated whether GA induced DNA damage and affected DNA repair-associated protein expression in human oral cancer SCC-4 cells. Flow cytometry assays were used to measure total viable cells and results indicated that GA decreased viable cells dose-dependently. The comet assay and 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) staining were used to measure DNA damage, as well as condensation and it was shown that GA induced DNA damage (comet tail) and DNA condensation in a dose-dependent manner. DNA gel electrophoresis was used to examine DNA fragmentation and we found that GA induced DNA ladder (fragmentation). Using western blotting it was shown that GA inhibited the protein expressions of MDC1, O(6)-methylguanine-DNA methyltransferase (MGMT), p-H2A.X, p53, DNA-dependent serine/threonine protein kinase (DNA-PK) and 14-3-3 proteins sigma (14-3-3σ) but increased p-p53, phosphate-ataxia-telangiectasia (p-H2A.X) and ataxia telangiectasia mutated and Rad3-related (p-ATR), phosphate-ataxia telangiectasia mutated (p-ATM) and breast cancer susceptibility protein 1 (BRCA1) in a 24-h treatment. The protein translocation was examined by confocal laser microscopy and results indicated that GA increased the levels of p-H2A.X, MDC1 and p-p53 in SCC-4 cells. In conclusion, we found that GA-induced cell death may proceed through the induced DNA damage and suppressed DNA repair-associated protein expression in SCC-4 cells.


Subject(s)
DNA Damage/drug effects , DNA Repair/genetics , Gallic Acid/administration & dosage , Mouth Neoplasms/drug therapy , Cell Line, Tumor , Cell Survival/drug effects , Cell Survival/genetics , DNA Repair/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Humans , Mouth Neoplasms/genetics , Mouth Neoplasms/pathology , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics
8.
In Vivo ; 29(2): 255-61, 2015.
Article in English | MEDLINE | ID: mdl-25792654

ABSTRACT

Polygonum cuspidatum is a traditional Chinese herbal medicine used in the treatment of various diseases. In the present study, we investigated whether the crude extract of Polygonum cuspidatum (CEPC) could affect immune responses of murine leukemia cells in vivo. Normal BALB/c mice were i.p. injected with WEHI-3 cells to generate leukemic mice and then were treated orally with CEPC at 0, 50, 100 and 200 mg/kg for three weeks. Animals were weighed and blood, liver, spleen samples were collected for further analyses. Results indicated that CEPC did not significantly affect the body and liver weight of animals, but reduced the weight of spleen when compared to control groups. Flow cytometric assay demonstrated that CEPC increased the percentage of CD3- (T-cell marker) and CD19- (B-cell marker) positive cells, but reduced that of CD11b-positive ones (monocytes). However, it did not significantly affect the proportion of Mac-3-positive cells (macrophages), compared to control groups. Results indicated that CEPC promoted phagocytosis by macrophages from blood samples at all examined doses but did not affect that of macrophages from the peritoneal cavity. CEPC also promoted natural killer cell activity of splenocytes at 200 mg/kg of CEPC. CEPC promoted B-cell proliferation at 200 mg/kg treatment when cells were stimulated with lipopolysaccharides but did not promote T-cell proliferation at three doses of CEPC treatment on concanavalin A stimulation.


Subject(s)
Fallopia japonica/chemistry , Killer Cells, Natural/drug effects , Killer Cells, Natural/immunology , Macrophages/drug effects , Macrophages/immunology , Phagocytosis/drug effects , Phagocytosis/immunology , Plant Extracts/pharmacology , Animals , Antigens, Surface/metabolism , Antineoplastic Agents, Phytogenic/administration & dosage , Antineoplastic Agents, Phytogenic/pharmacology , B-Lymphocytes/drug effects , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , Cell Line, Tumor , Cytotoxicity, Immunologic/drug effects , Drugs, Chinese Herbal/administration & dosage , Drugs, Chinese Herbal/pharmacology , Leukemia, Experimental , Lymphocyte Activation/drug effects , Lymphocyte Activation/immunology , Male , Mice , Plant Extracts/administration & dosage , T-Lymphocytes/drug effects , T-Lymphocytes/immunology , T-Lymphocytes/metabolism
9.
Anticancer Res ; 35(2): 795-804, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25667459

ABSTRACT

Cantharidin is an active component of mylabris, which has been used as a traditional Chinese medicine. Cantharidin has been shown to have antitumor activity against several types of human cancers in vitro and in animal models in vivo. We investigated whether cantharidin induces DNA damage and affects DNA damage repair-associated protein levels in TSGH8301 human bladder cancer cells. Using flow cytometry to measure viable cells, cantharidin was found to reduce the number of viable cells in a dose-dependent manner. Comet assay, 4',6-diamidino-2-phenylindole (DAPI) staining and DNA gel electrophoresis were used to measure DNA damage and condensation; the results indicated that cantharidin induced DNA damage (comet tail), DNA condensation (white DAPI staining) and DNA damage (DNA smear). Results from western blotting showed that cantharidin inhibited the expression of DNA-dependent serine/threonine protein kinase, poly-ADP ribose polymerase, phosphate-ataxia-telangiectasia and RAD3-related, O-6-methylguanine-DNA methyltransferase, breast cancer susceptibility protein 1, mediator of DNA damage checkpoint protein 1, phospho-histone H2A.X, but increased that of phosphorylated p53 following 6 and 24 h treatment. Confocal laser microscopy was used to examine the protein translocation; cantharidin suppressed the levels of p-H2A.X and MDC1 but increased the levels of p-p53 in TSGH8301 cells. In conclusion, we found that cantharidin-induced cell death may occur through the induction of DNA damage and suppression of DNA repair-associated protein expression in TSGH8301 cells.


Subject(s)
Cantharidin/pharmacology , DNA Damage , DNA Repair , Neoplasm Proteins/antagonists & inhibitors , Urinary Bladder Neoplasms/pathology , Cell Line, Tumor , Humans , Microscopy, Confocal , Neoplasm Proteins/metabolism , Urinary Bladder Neoplasms/genetics , Urinary Bladder Neoplasms/metabolism
10.
Environ Toxicol ; 30(11): 1343-53, 2015 Nov.
Article in English | MEDLINE | ID: mdl-24890016

ABSTRACT

Diallyl trisulfide (DATS), a chemopreventive dietary constituent and extracted from garlic, has been shown to against cultured many types of human cancer cell liens but the fate of apoptosis in murine leukemia cells in vitro and immune responses in leukemic mice remain elusive. Herein, we clarified the actions of DATS on growth inhibition of murine leukemia WEHI-3 cells in vitro and used WEHI-3 cells to generate leukemic mice in vivo, following to investigate the effects of DATS in animal model. In in vitro study, DATS induced apoptosis of WEHI-3 cells through the G0/G1 phase arrest and induction of caspase-3 activation. In in vivo study DATS decreased the weight of spleen of leukemia mice but did not affect the spleen weight of normal mice. DATS promoted the immune responses such as promotions of the macrophage phagocytosis and NK cell activities in WEHI-3 leukemic and normal mice. However, DATS only promotes NK cell activities in normal mice. DATS increases the surface markers of CD11b and Mac-3 in leukemia mice but only promoted CD3 in normal mice. In conclusion, the present study indicates that DATS induces cell death through induction of apoptosis in mice leukemia WHEI-3 cells. DATS also promotes immune responses in leukemia and normal mice in vivo.


Subject(s)
Allyl Compounds/pharmacology , Anticarcinogenic Agents/pharmacology , Apoptosis/drug effects , Leukemia, Experimental/immunology , Leukemia, Experimental/prevention & control , Sulfides/pharmacology , Allyl Compounds/therapeutic use , Animals , Anticarcinogenic Agents/therapeutic use , Antigens, Differentiation/immunology , Caspase 3/metabolism , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Cytotoxicity, Immunologic/drug effects , Garlic/chemistry , Killer Cells, Natural/drug effects , Killer Cells, Natural/immunology , Lymphocyte Activation/drug effects , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/immunology , Mice , Mice, Inbred BALB C , Neoplasm Transplantation , Phagocytosis/drug effects , Phagocytosis/immunology , Spleen/drug effects , Spleen/immunology , Sulfides/therapeutic use
11.
Molecules ; 19(10): 16588-608, 2014 Oct 15.
Article in English | MEDLINE | ID: mdl-25322282

ABSTRACT

Osteosarcoma is the most common malignant primary bone tumor in children and young adults and lung metastasis is the main cause of death in those patients. Deguelin, a naturally occurring rotenoid, is known to be an Akt inhibitor and to exhibit cytotoxic effects, including antiproliferative and anticarcinogenic activities, in several cancers. In the present study, we determined if deguelin would inhibit migration and invasion in U-2 OS human osteosarcoma cells. Deguelin significantly inhibited migration and invasion of U-2 OS human osteosarcoma cells which was associated with a reduction of activities of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9). Furthermore, results from western blotting indicated that deguelin decreased the cell proliferation and cell growth-associated protein levels, such as SOS1, PKC, Ras, PI3K, p-AKT(Ser473), IRE-1α, MEKK3, iNOS, COX2, p-ERK1/2, p-JNK1/2, p-p38; the cell motility and focal adhesion-associated protein levels, such as Rho A, FAK, ROCK-1; the invasion-associated protein levels, such as TIMP1, uPA, MMP-2. MMP-9, MMP-13, MMP-1 and VEGF in U-2 OS cells. Confocal microscopy revealed that deguelin reduced NF-κB p65, Rho A and ROCK-1 protein levels in cytosol. MMP-7, MMP-9 and Rho A mRNA levels were suppressed by deguelin. These in vitro results provide evidence that deguelin may have potential as a novel anti-cancer agent for the treatment of osteosarcoma and provides the rationale for in vivo studies in animal models.


Subject(s)
Bone Neoplasms/metabolism , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Osteosarcoma/metabolism , Rotenone/analogs & derivatives , Bone Neoplasms/pathology , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Humans , In Vitro Techniques , Neoplasm Invasiveness , Osteosarcoma/pathology , Rotenone/pharmacology
12.
Oncol Rep ; 32(1): 355-61, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24859325

ABSTRACT

Oral cancer is one of the major causes of mortality in humans and squamous cell carcinoma is the most common type of oral cancer. Gallic acid (GA) is a natural product that induces cell death through cell cycle arrest and induction of apoptosis. There is no available information on whether GA affects cell migration and invasion of human oral cancer cells. We determined if GA inhibited migration and invasion of SCC-4 (human squamous cell carcinoma) human oral cancer cells. GA significantly inhibited migration and invasion of SCC-4 cells based on results from the wound healing assay and Matrigel Cell Migration Assay and Invasion System. We also showed that GA significantly inhibited matrix metalloproteinase (MMP)-2 and MMP-9 activity. GA reduced protein levels of FAK, MEKK3, p-PERK, p-p38, p-JNK1/2, p-ERK1/2, SOS1, RhoA, Ras, PKC, p-AKT(Thr308), PI3K, NF-κB p65, MMP-2 and MMP-9 in SCC-4 cells. Translocation of NF-κB and RhoA from the cytosol to the nucleus was reduced by GA in SCC-4 cells. In summary, GA inhibits migration and invasion of SCC-4 cells by inhibiting NF-κB expression causing suppression of MMP-2 and MMP-9 activity. GA may have potential as a therapeutic agent for the treatment of oral cancer.


Subject(s)
Gallic Acid/pharmacology , Mouth Neoplasms/pathology , Neoplasm Invasiveness/pathology , Signal Transduction/drug effects , Cell Line, Tumor , Cell Movement/drug effects , Cell Survival/drug effects , Humans , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , NF-kappa B/metabolism , Protein Transport/drug effects , ras Proteins/metabolism
13.
Environ Toxicol ; 29(9): 969-80, 2014 Sep.
Article in English | MEDLINE | ID: mdl-23315830

ABSTRACT

Crude extract of Rheum palmatum L (CERP) has been used to treat different diseases in the Chinese population for decades. In this study, we investigated the effects of CERP on LS1034 human colorectal cancer cells in vitro and also examined possible mechanisms of cell death. Flow cytometric assays were used to measure the percentage of viable cells, cell cycle distribution including the sub-G1 phase (apoptosis), the activities of caspase-8, -9, and -3, reactive oxygen species (ROS) and Ca(2+) levels, and mitochondrial membrane potential (ΔΨm). DNA damage, nuclei condensation, protein expression, and translocation were examined by Comet assay, 4'-6-diamidino-2-phenylindole (DAPI) staining, Western blotting, and confocal laser system microscope, respectively. CERP induced apoptosis as seen by DNA fragmentation and DAPI staining in a concentration- and time-dependent manner in cancer cells. CERP was associated with an increase in the Bax/Bcl-2 protein ratio and CERP promoted the activities of caspase-8, -9, and -3. Both ROS and Ca(2+) levels were increased by CERP but the compound decreased levels of ΔΨm in LS1034 cells. Laser confocal microscope also confirmed that CERP promoted the expressions of AIF, Endo G, cytochrome c, and GADD153 to induce apoptosis through mitochondrial-dependent pathway.


Subject(s)
Apoptosis/drug effects , Caspases/metabolism , Colonic Neoplasms/pathology , Plant Extracts/pharmacology , Rheum/chemistry , Calcium/metabolism , Cell Cycle/drug effects , Cell Line, Tumor , Colonic Neoplasms/enzymology , Comet Assay , Cytochromes c/metabolism , DNA Damage , Humans , Indoles/chemistry , Membrane Potential, Mitochondrial/drug effects , Protein Transport , Reactive Oxygen Species/metabolism , bcl-2-Associated X Protein/metabolism
14.
Anticancer Res ; 33(5): 1941-50, 2013 May.
Article in English | MEDLINE | ID: mdl-23645742

ABSTRACT

Quercetin, a principal flavanoid compound in onions, has been shown to possess a wide spectrum of pharmacological properties, including anticancer activities. Our earlier study showed that quercetin induced cytotoxic effects on SAS human oral cancer cells. In this study, we found that quercetin significantly reduced wound closure of SAS cells in culture plates after 12- and 24-h treatments. Results indicated that quercetin inhibited the expression and activity of matrix metalloproteinase (MMP)-2 and MMP-9, as measured by western blotting and gelatin zymography. The results from western blotting also showed that quercetin reduced the protein levels of MMP-2, -7, -9 and -10, vascular endothelial growth factor (VEGF), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65, inductible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), urokinase-type plasminogen activator (uPA), phosphatidylinositide-3 kinases (PI3K), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (IKBα), IKB-α/ß, phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor kinase, alpha/beta (p-IKKα/ß), focal adhesion kinase (FAK), son of sevenless homolog-1 (SOS1), growth factor receptor-bound protein-2 (GRB2), mitogen-activated protein kinase kinase kinase-3 (MEKK3), MEKK7, extracellular-signal-regulated kinase 1/2 (ERK1/2), p-ERK1/2, c-Jun N-terminal kinase 1/2 (JNK1/2), p38, p-p38, Jun proto-oncogene (c-JUN) and p-c-JUN but it did not affect Ras homolog gene family, member A (RhoA), Protein kinase C (PKC) and rat sarcoma viral oncogene homolog (RAS) in SAS cells. Confocal laser microscopy also showed that quercetin promoted the expressions of RhoA and Rho-associated, coiled-coil containing protein kinase-1 (ROCK1), but inhibited the expression of NF-κB p65 in SAS cells. It is concluded from these data that inhibition of migration and invasion of SAS cells by quercetin is associated with the down-regulation of PKC and RhoA by blocking MAPK and PI3K/AKT signaling pathways and NF-κB and uPA, resulting in inhibition of MMP-2 and MMP-9 signaling.


Subject(s)
Cell Movement/drug effects , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Mouth Neoplasms/pathology , NF-kappa B/metabolism , Quercetin/pharmacology , Signal Transduction/drug effects , Antioxidants/pharmacology , Apoptosis/drug effects , Blotting, Western , Cell Adhesion/drug effects , Cell Proliferation/drug effects , Cyclooxygenase 2/metabolism , Humans , Immunoenzyme Techniques , MAP Kinase Signaling System/drug effects , Mitogen-Activated Protein Kinase 3/metabolism , Mouth Neoplasms/drug therapy , Mouth Neoplasms/metabolism , Neoplasm Invasiveness , Phosphatidylinositol 3-Kinases/metabolism , Protein Transport , Proto-Oncogene Mas , Tumor Cells, Cultured
15.
Article in English | MEDLINE | ID: mdl-23431332

ABSTRACT

Cancer metastasis becomes an initial cause of cancer death in human population. In many cancers, it has been shown that the high levels of matrix metalloproteinase (MMP)-2 and/or MMP-9 are associated with the invasive phenotypes of cancer cells. In this study, we investigated the effects of cantharidin, a derivative of blister beetles which is one of the traditional Chinese medicines, on the adhesion, migration, and invasion of human bladder cancer TSGH-8301 cells. Cantharidin effectively suppressed TSGH-8301 cell adhesion, migration, and invasion in a concentration-dependent manner. Results from Western blotting, RT-PCR, and gelatin zymography assays indicated that cantharidin blocked the protein levels, gene expression (mRNA), and activities of MMP-2 and -9 in TSGH-8301 cells. Cantharidin also significantly suppressed the protein expressions of p-p38 and p-JNK1/2 in TSGH-8301 cells. Taken together, cantharidin was suggested to present antimetastatic potential via suppressing the levels of MMP-2 and MMP-9 expression that might be mediated by targeting the p38 and JNK1/2 MAPKs pathway in TSGH-8301 human bladder cancer cells.

16.
Environ Toxicol ; 28(3): 127-36, 2013 Mar.
Article in English | MEDLINE | ID: mdl-21626647

ABSTRACT

Enhanced cruciferous vegetable consumption is associated with the reduction of cancer incidence as shown in epidemiological studies. Phenethyl isothiocyanate (PEITC), one of the important compounds in cruciferous vegetables, has been shown to induce apoptosis in many types of human cancer cell lines, but there is no available information addressing the effects on normal and leukemia mice in vivo. The purpose of this study is to focus on the in vivo effects of PEITC on immune responses of normal and WEHI-3 leukemia BALB/c mice in vivo. Influences of PEITC on BALB/c mice after intraperitoneal (i.p.) injection with WEHI-3 cells and normal mice were investigated. In normal BALB/c mice, PEITC did not affect the body weight when compared to the olive oil treated animals. Moreover, PEITC promoted phagocytosis by macrophages from peripheral blood mononuclear cells (PBMC) and peritoneal cavity, increased the levels of CD11b and Mac-3, decreased the level of CD19 and promoted natural killer (NK) cell cytotoxic activity, but it did not alter the level of CD3. Also, PEITC enhanced T cell proliferation after concanavalin A (Con A) stimulation. Otherwise, PEITC increased the body weight, but decreased the weight of liver and spleen as compared to the olive oil-treated WEHI-3 leukemia mice. PEITC also increased the level of CD19, decreased the levels of CD3 and Mac-3 rather than influence in the level of CD11b, suggesting that the differentiation of the precursor of macrophages and T cells was inhibited, but the differentiation of the precursor of B cells was promoted in leukemia mice. Furthermore, PEITC enhanced phagocytosis by monocytes and macrophages from PBMC and peritoneal cavity, and also promoted the NK cell cytotoxic activity in comparison with the group of leukemia mice. Based on these observations, the biological properties of PEITC can promote immune responses in normal and WEHI-3 leukemia mice in vivo. © 2011 Wiley Periodicals, Inc. Environ Toxicol, 2013.


Subject(s)
Isothiocyanates/pharmacology , Leukemia, Experimental/drug therapy , Leukemia, Experimental/immunology , Animals , B-Lymphocytes/drug effects , B-Lymphocytes/immunology , Body Weight/drug effects , Cell Line, Tumor , Concanavalin A/pharmacology , Cytotoxicity, Immunologic/drug effects , Immunomodulation/drug effects , Killer Cells, Natural/drug effects , Killer Cells, Natural/immunology , Leukemia, Experimental/blood , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/immunology , Liver/drug effects , Liver/immunology , Lymphocyte Activation/drug effects , Macrophages/drug effects , Macrophages/immunology , Male , Mice , Mice, Inbred BALB C , Phagocytosis/drug effects , Phagocytosis/immunology , Spleen/drug effects , Spleen/immunology , T-Lymphocytes/drug effects , T-Lymphocytes/immunology
17.
J Agric Food Chem ; 60(45): 11395-402, 2012 Nov 14.
Article in English | MEDLINE | ID: mdl-23066961

ABSTRACT

The cytostatic drug from natural products has acted as a chemotherapeutic agent used in treatment of a wide variety of cancers. Apigenin, a type of flavonoid, exhibits anticancer actions, but there is no report to show that apigenin induced apoptosis in osteosarcoma cells. The aim of this study was to investigate the effects of apigenin on U-2 OS human osteosarcoma cells and clarify that the apigenin-induced apoptosis-associated signals. The cytotoxic effects of apigenin were examined by culturing U-2 OS cells with or without apigenin. The percentage of viable cells via PI staining, apoptotic cells, productions of ROS and Ca²âº, and the level of mitochondrial membrane potential (ΔΨm) were assayed by flow cytometry. The levels of apoptosis-related proteins were measured by immunoblotting. Results indicated that apigenin significantly decreased cell viability. Apigenin effectively induced apoptosis through the activations of caspase-3, -8, -9, and BAX and promoted the release of AIF in U-2 OS cells. In nude mice bearing U-2 OS xenograft tumors, apigenin inhibited tumor growth. In conclusion, apigenin has anticancer properties for induction of cell apoptosis in U-2 OS cells and suppresses the xenograft tumor growth. These findings offer novel information that apigenin possibly possesses anticancer activity in human osteosarcoma.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apigenin/pharmacology , Apoptosis/drug effects , Bone Neoplasms/drug therapy , Cell Proliferation/drug effects , Mitochondria/metabolism , Osteosarcoma/drug therapy , Animals , Bone Neoplasms/metabolism , Bone Neoplasms/physiopathology , Cell Line, Tumor , Cell Survival/drug effects , Down-Regulation/drug effects , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Mitochondria/drug effects , Osteosarcoma/metabolism , Osteosarcoma/physiopathology , Xenograft Model Antitumor Assays
18.
Article in English | MEDLINE | ID: mdl-22919418

ABSTRACT

Phenethyl isothiocyanate (PEITC), an effective anticancer and chemopreventive agent, has been reported to inhibit cancer cell growth through cell-cycle arrest and induction of apoptotic events in various human cancer cells models. However, whether PEITC inhibits human oral squamous cell carcinoma HSC-3 cell growth and its underlying mechanisms is still not well elucidated. In the present study, we evaluated the inhibitory effects of PEITC in HSC-3 cells and examined PEITC-modulated cell-cycle arrest and apoptosis. The contrast-phase and flow cytometric assays were used for examining cell morphological changes and viability, respectively. The changes of cell-cycle and apoptosis-associated protein levels were determined utilizing Western blotting in HSC-3 cells after exposure to PEITC. Our results indicated that PEITC effectively inhibited the HSC-3 cells' growth and caused apoptosis. PEITC induced G(0)/G(1) phase arrest through the effects of associated protein such as p53, p21, p17, CDK2 and cyclin E, and it triggered apoptosis through promotion of Bax and Bid expression and reduction of Bcl-2, leading to decrease the levels of mitochondrial membrane potential (ΔΨ(m)), and followed the releases of cytochrome c, AIF and Endo G then for causing apoptosis in HSC-3 cells. These results suggest that PEITC could be an antitumor compound for oral cancer therapy.

19.
Anticancer Res ; 32(7): 2895-903, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22753753

ABSTRACT

Anti-metastasis by reducing cellular migration and invasion and by deregulating the expression of matrix metalloproteinases (MMPs) is a therapeutic approach for cancer treatment. The objective of this study focused on the effects of the novel compound 6-pyrrolidinyl-2-(2-hydroxyphenyl)-4-quinazolinone (MJ-29) regarding anti-metastatic actions on human oral squamous cell carcinoma CAL 27 cells and on the verification of the underlying related molecular mechanisms of this event. MJ-29 concentration- and time-dependently caused a suppression of cell adhesive ability utilizing cell adhesion assay; it also inhibited the migration and invasion of CAL 27 cells using scratch wound closure and transwell invasion assays in a concentration-dependent response. Importantly, we confirmed that the applied concentration range of MJ-29 exhibited no dramatic influence of cytotoxicity on CAL 27 cells using the thiazolyl blue tetrazolium bromide assay. MJ-29 also attenuated the enzymatic activity of MMP-2 and MMP-9. Furthermore, we found that activation of their upstream protein kinases, by MJ-29, potentially exerted an inhibitory effect on the phosphorylated protein levels of extracellular regulated protein kinase 1/2, p38 and c-Jun N-terminal kinase 1/2, as well as serine/threonine kinase AKT by MJ-29 in CAL 27 cells. The expression of RAS and focal adhesion kinase was also down-regulated in MJ-29-treated CAL 27 cells. Collectively, these findings provide further evidence for the molecular signaling basis of the effects of MJ-29 on suppression of migration and invasion which might be useful as a therapeutic strategy to treat human oral cancer.


Subject(s)
MAP Kinase Signaling System/drug effects , Matrix Metalloproteinase 2/biosynthesis , Matrix Metalloproteinase 9/biosynthesis , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Mouth Neoplasms/drug therapy , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Quinazolinones/pharmacology , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/enzymology , Carcinoma, Squamous Cell/pathology , Cell Adhesion/drug effects , Cell Line, Tumor , Cell Movement/drug effects , Down-Regulation/drug effects , Head and Neck Neoplasms/drug therapy , Head and Neck Neoplasms/enzymology , Head and Neck Neoplasms/pathology , Humans , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Matrix Metalloproteinase Inhibitors , Mitogen-Activated Protein Kinases/metabolism , Mouth Neoplasms/enzymology , Mouth Neoplasms/genetics , Mouth Neoplasms/pathology , Neoplasm Invasiveness , Proto-Oncogene Proteins c-akt/metabolism , Pyrrolidines/pharmacology , Squamous Cell Carcinoma of Head and Neck
20.
In Vivo ; 26(4): 665-70, 2012.
Article in English | MEDLINE | ID: mdl-22773581

ABSTRACT

Chrysin (5,7-dihydroxyflavone), a natural and biologically active flavonoid found in plants, possesses many biological activities and anticancer effects. However, there is no available evidence regarding the antileukemia responses to chrysin in a mouse model. We hypothesized that chrysin affects murine WEHI-3 leukemia cells in vitro and in vivo. The present study showed that chrysin at concentrations of 5-50 µM reduced the cell viability in concentration- and time-dependent manners. In an in vivo study, WEHI-3 leukemic BALB/c mice were established in order to determine antileukemia activity of chrysin. Our results revealed that chrysin increased the percentage of CD3 (T-cell maker), CD19 (B-cell maker) and Mac-3 (macrophages) cell surface markers in treated mice as compared with the untreated leukemia group. However, chrysin did not significantly influence the level of CD11b (a monocyte maker) in treated mice. Moreover, there was a significant increase in phagocytosis by macrophages from peripheral blood mononuclear cells, but no effect in those from the peritoneal cavity in leukemic mice after chrysin treatment. Isolated splenocytes from chrysin-treated leukemic mice demonstrated an increase of natural killer (NK) cell cytotoxicity. Based on these observations, chrysin might exhibit antileukemia effects on a murine WEHI-3 cell line-induced leukemia in vivo.


Subject(s)
B-Lymphocytes/drug effects , Disease Models, Animal , Flavonoids/pharmacology , Killer Cells, Natural/drug effects , Leukemia, Experimental/pathology , Macrophages/drug effects , Phagocytosis , T-Lymphocytes/drug effects , Animals , B-Lymphocytes/immunology , Cell Line, Tumor , Cytotoxicity, Immunologic , Killer Cells, Natural/immunology , Leukemia, Experimental/immunology , Macrophages/immunology , Male , Mice , Mice, Inbred BALB C , T-Lymphocytes/immunology
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