1.RT-PCR of Up-Regulated Factors in Abnormally Proliferated Vascular Endothelial Cells by 1,2- Dimethylhydrazine.
Sung Ho KIM ; Young Seok KANG ; Yong Chan BAE ; Suk Young PARK ; Su Bong NAM
Journal of the Korean Society of Plastic and Reconstructive Surgeons 2005;32(6):689-698
Many studies for verifying angiogenesis have been in progress, especially in the field of abnormal vascular proliferation to explain the pathogenesis and to develop a treatment of several diseases. In our previous experiments, endothelial cell proliferations were induced by DMH stimulation in vitro, and the 177 factors(142 up- regulated and 35 down-regulated factors) were identified. Among the up-regulated factors, 9 substances (EFEMP1, CTGF, CYR61, ITGbeta1, FHL2, SERPINE1, MYC, PTTG1 and MSH6) were selected, which were related to cell proliferation and showed high signal intensities. The RNA was isolated from HUVECs at the time of 0, 6, 12, 24 hours after the DMH treatment, and RNA of control group HUVECs was also isolated. Genetic information of selected molecules was used to make primer for each, and RT-PCR was performed to analyze both groups. In control and treatment groups, each substance presented variety of manifestation degree according to time differences. EFEMP1, CTGF, CYR61, ITGbeta1, FHL2 and MYC were related to abnormal vascular proliferation steadily and SERPINE1, PTTG1 and MSH6 were related secondarily. CTGF was related to both normal and abnormal proliferation, but it played a more significant role in abnormal proliferation from earlier stage. EFEMP1, CYR61, ITGbeta1, FHL2 and MYC were similar to CTGF, although the relation appeared lately. Further study should be performed to analyze the expressions and the interactions of growth factors, which could be utilized in the new therapeutic development.
Cell Proliferation
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Dimenhydrinate
;
Dimethylhydrazines
;
Endothelial Cells*
;
Intercellular Signaling Peptides and Proteins
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RNA
;
Umbilical Veins
2.The effect of vitamine E on intestinal tumorigenesis by 1,2-dimethylhydrazine.
Kang Hyuk LEE ; Dae Young KANG
Journal of the Korean Cancer Association 1991;23(1):29-38
No abstract available.
1,2-Dimethylhydrazine*
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Carcinogenesis*
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Vitamins*
3.Gene Expression Profiling of 1,2-Dimethylhydrazine-Stimulated Human Umbilical Vein Endothelial Cells.
Hyun Ok KIM ; Young Seok KANG ; Yong Chan BAE ; Suk Young PARK ; So Min HWANG ; Su Bong NAM
Journal of the Korean Society of Plastic and Reconstructive Surgeons 2004;31(6):858-864
The purposes of this study are to establish standard model in which endothelial cell proliferations are induced by DMH stimulation in vitro, and to analyze the gene expressions of proliferative HUVECs using DNA chip technique which could evaluate the mechanisms of angiogenesis, and the development of vascular tumors. To perform the MTT assay in 96-well microplates, 104 cells were seeded in each well which were cultured in medium. On the third day, the cells were treated with 5 different concentrations of diluted DMH from 10 to 10-3 ng/ml. Five DMH-treated groups were compared with the control group which was not treated with DMH. The optical densities in each group were measured at the time of 0, 6, 12, 24, 36, 48, and 72 hours after DMH treatment. The same experiment was repeated 9 times. Statistically significant cell proliferations were observed in 1 and 10-1ng/ml group. The RNAs were isolated from HUVECs of control group and 1ng/ml DMH-treated group, and they were used to analyze the gene expressions using DNA chip technique. One hundred and seventy-seven genes(142 of up-retulated genes and 35 down-regulated genes) were identified, and several genes were associated with VEGF and FGF production. Also DMH could affect expression of genes that involve oncogenesis. Further study should be performed to evaluate the processes of angiogenesis and morphogenesis of vascular tumors, which could be utilized in the development of new therapeutic approaches.
Carcinogenesis
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Dimenhydrinate
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Dimethylhydrazines
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Endothelial Cells
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Gene Expression Profiling*
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Gene Expression*
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Human Umbilical Vein Endothelial Cells*
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Humans*
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Morphogenesis
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Oligonucleotide Array Sequence Analysis
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RNA
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Umbilical Veins
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Vascular Endothelial Growth Factor A
4.Effects of astragalosides on induction of colorectal aberrant crypt foci by dimethylhydrazine and metabolizing enzymes in liver microsomes in rats.
Journal of Southern Medical University 2010;30(7):1720-1723
OBJECTIVETo observe the effects of astragalosides on content of rat liver microsomal cytochrome P450 (CYP450), activity of glutathione-S-transferase (GST) and dimethylhydrazine (DMH)-induced aberrant crypt foci formation in rats.
METHODSForty SD rats were randomly and equally divided into control group, Astragalus group, DMH group, Astragalus+DMH group. The animals were killed by off neck and the colorectal and liver tissues taken after treatment with astragalosides and dimethyl hydrazine . The colorectal tissues were stained by methylene blue, ACFs observed and counted, and liver microsomes isolated by differential centrifugation. The total enzyme content was detected by using differential spectrometry for carbon monoxide reduction. The glutathione (GSH) level was detected by using spectrophotometry to reflect the activity of the GST.
RESULTSThe numbers of ACF and large ACF in Astragalus+DMH group were more significantly decreased than the DMH group (P<0.05). Compared with the control group, Astragalus group and Astragalus+DMH group, CYP450 level was decreased significantly, and GST activity increased significantly in DMH group (P<0.05).
CONCLUSIONAstragalosides might reduce the number of colorectal aberrant crypt foci induced by dimethylhydrazine possibly by reducing the content of hepatic CYP450 and increasing GST activity in rats.
Aberrant Crypt Foci ; metabolism ; Animals ; Colorectal Neoplasms ; Dimethylhydrazines ; toxicity ; Male ; Microsomes, Liver ; drug effects ; enzymology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology
5.Effect of jinfu kang to experimental precancerous colon lesions and urinary metabolites in rat.
Yin-ping QIU ; Ming-ming SU ; Da-zheng WU ; Ai-hua ZHAO ; Yu-min LIU ; Wei JIA
China Journal of Chinese Materia Medica 2008;33(22):2653-2657
OBJECTIVE: To profile urinary metabolite variations from 1, 2-dimethylhydrazine (DMH)-induced precancerous colon rats, Jinfu Kang treated rats and healthy controls.
METHODWe used ethyl chloroformate derivatization and gas chromatography-mass spectrometry (GC-MS) based metabonomic method to analyze rat urines.
RESULTThe time-dependent variations of metabolite profile showed a progressive deviation of the metabolism in the model group from the initial pattern over time and a systemic recovery of the metabolism in the treatment group, which is consistent with the histological results. The in-depth analysis indicated that the disorder of tricarboxylic acid cycle (TCA), tryptophan metabolism, polyamine metabolism and gut flora structure were associated with DMH intervention.
CONCLUSIONMetabolic study revealed that Jinfu Kang can effectively reverse metabolic departures in DMH-induced precancerous colon rat, which is consistent with pathological results.
Animals ; Colonic Neoplasms ; chemically induced ; pathology ; Colonic Polyps ; chemically induced ; drug therapy ; urine ; Dimethylhydrazines ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Gas Chromatography-Mass Spectrometry ; Male ; Rats ; Rats, Wistar
6.Effects of Coptis chinensis and Evodia rutaecarpa water extract on DMH-induced precancerous lesion of colon.
Li DONG ; Hailian SHI ; Guang JI ; Dazheng WU
China Journal of Chinese Materia Medica 2010;35(9):1185-1188
OBJECTIVETo investigate the effects of Coptis chinensis and Evodia rutaecarpa water extract on precancerous lesion of colon induced by DMH and proliferation and apoptosis changes of colon mucosa crypts.
METHODPrecancerous lesion of colon was induced by DMH. The changes of proliferation and apoptosis of colon mucosa crypts were detected by morphological analysis. The numbers of aberrant crypt foci (ACF) were measured by feulgen staining.
RESULTC. chinensis and E. rutaecarpa water extract could significantly inhibit the formation of ACF in model animals. The proliferative crypts were increased obviously in middle and distal colon, and decreased by C. chinensis and E. rutaecarpa water extract. The apoptosis crypts were increased in distal colon but not middle colon. C. chinensis and E. rutaecarpa water extract could promote apoptosis of both middle and distal colon.
CONCLUSIONC. chinensis and E. rutaecarpa water extract could significantly inhibit the formation of ACF in model animals. These results indicated that C. chinensis and E. rutaecarpa water extract maybe have an inhibitory and clinically therapeutic effect on colon cancer, which were partly resulted from inhibiting proliferation and promoting apoptosis of crypts in middle and distal colon.
Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; chemically induced ; drug therapy ; pathology ; physiopathology ; Coptis ; chemistry ; Dimethylhydrazines ; adverse effects ; Disease Models, Animal ; Evodia ; chemistry ; Humans ; Male ; Plant Extracts ; administration & dosage ; Random Allocation ; Rats ; Rats, Wistar
7.Triptolide downregulates Rac1 and the JAK/STAT3 pathway and inhibits colitis-related colon cancer progression.
Zhipeng WANG ; Haifeng JIN ; Ruodan XU ; Qibing MEI ; Daiming FAN
Experimental & Molecular Medicine 2009;41(10):717-727
Triptolide, a diterpenoid triepoxide from the traditional Chinese medicinal herb Tripterygium wilfordii Hook. f., is a potential treatment for autoimmune diseases as well a possible anti-tumor agent. It inhibits proliferation of coloretal cancer cells in vitro and in vivo. In this study, its ability to block progress of colitis to colon cancer, and its molecular mechanism of action are investigated. A mouse model for colitis-induced colorectal cancer was used to test the effect of triptolide on cancer progression. Treatment of mice with triptolide decreased the incidence of colon cancer formation, and increased survival rate. Moreover, triptolide decreased the incidence of tumors in nude mice inoculated with cultured colon cancer cells dose-dependently. In vitro, triptolide inhibited the proliferation, migration and colony formation of colon cancer cells. Secretion of IL6 and levels of JAK1, IL6R and phosphorylated STAT3 were all reduced by triptolide treatment. Triptolide prohibited Rac1 activity and blocked cyclin D1 and CDK4 expression, leading to G1 arrest. Triptolide interrupted the IL6R-JAK/STAT pathway that is crucial for cell proliferation, survival, and inflammation. This suggests that triptolide might be a candidate for prevention of colitis induced colon cancer because it reduces inflammation and prevents tumor formation and development.
Animals
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Cell Transformation, Neoplastic/*drug effects
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Colitis/complications
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Colonic Neoplasms/chemically induced/*drug therapy/metabolism/pathology
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Dextran Sulfate/toxicity
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Dimethylhydrazines/toxicity
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Diterpenes/*administration & dosage
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Epoxy Compounds/administration & dosage
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Humans
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Interleukin-6/biosynthesis
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Janus Kinases/metabolism
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Male
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Mice
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Mice, Inbred BALB C
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Mice, Inbred ICR
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Mice, Nude
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Neoplasm Transplantation
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Phenanthrenes/*administration & dosage
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STAT3 Transcription Factor/metabolism
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Signal Transduction/*drug effects
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Tumor Burden/drug effects
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rac1 GTP-Binding Protein/*biosynthesis
8.Chemopreventive and metabolic effects of inulin on colon cancer development.
Emilia HIJOVA ; Viktoria SZABADOSOVA ; Jana STOFILOVA ; Gabriela HRCKOVA
Journal of Veterinary Science 2013;14(4):387-393
Prebiotics modulate microbial composition and ensure a healthy gastrointestinal tract environment that can prevent colon cancer development. These natural dietary compounds are therefore potential chemopreventive agents. Thirty Sprague-Dawley rats (4 months old) were experimentally treated with procarcinogen dimethylhydrazine to induce colon cancer development. The rats were randomly assigned to three groups: a control group (CG), a group treated with dimethylhydrazine (DMH), and a group given DMH and inulin, a prebiotic (DMH+PRE). The effects of inulin on the activities of bacterial glycolytic enzymes, short-chain fatty acids, coliform and lactobacilli counts, cytokine levels, and cyclooxygenase-2 (COX-2) and transcription nuclear factor kappa beta (NFkappaB) immunoreactivity were measured. Inulin significantly decreased coliform counts (p < 0.01), increased lactobacilli counts (p < 0.001), and decreased the activity of beta-glucuronidase (p < 0.01). Butyric and propionic concentrations were decreased in the DMH group. Inulin increased its concentration that had been reduced by DMH. Inulin decreased the numbers of COX-2- and NFkappaB-positive cells in the tunica mucosae and tela submucosae of the colon. The expression of IL-2, TNFalpha, and IL-10 was also diminished. This 28-week study showed that dietary intake of inulin prevents preneoplastic changes and inflammation that promote colon cancer development.
Animals
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Bacterial Proteins/genetics/metabolism
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Colon/enzymology
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Colonic Neoplasms/chemically induced/*drug therapy/metabolism
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Colony Count, Microbial
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Cyclooxygenase 2/genetics/metabolism
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Cytokines/blood/genetics
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Diet
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Dietary Supplements/analysis
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Dimethylhydrazines/toxicity
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Enterobacteriaceae/drug effects/physiology
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Fatty Acids, Volatile/genetics/metabolism
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Female
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Gene Expression Regulation/drug effects
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Inulin/administration & dosage/*metabolism
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Lactobacillaceae/drug effects/physiology
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Male
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NF-kappa B/genetics/metabolism
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Prebiotics/*analysis
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Rats
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Rats, Sprague-Dawley
9.Sequential Changes in Aberrant Crypt Foci and Lectin Expression in the Early and Late Stages of DMH-Induced Colon Carcinogenesis in Rats.
Hye Sung WON ; Lee So MAENG ; Hiun Suk CHAE ; Hyung Keun KIM ; Young Suk CHO ; Jin Hyoung KANG ; Hong Seok JANG ; Mi Ryeong RYU
Gut and Liver 2012;6(2):229-234
BACKGROUND/AIMS: The purpose of this study was to investigate the malignant potential of aberrant crypt foci (ACF) by measuring the multiplicity of crypts and lectin expression in the early and late stages of 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis. METHODS: Six-week-old Wistar rats were injected subcutaneously with DMH for 27 weeks. We classified ACF according to the number of crypts per ACF as a few crypts (< or =3 crypts, FC ACF) or numerous crypts (> or =4 crypts, NC ACF). Immunohistochemistry was used to evaluate lectin expression. RESULTS: In the early stage, FC ACF (590/1,902, 31.0%) occurred more frequently than NC ACF (35/449, 7.8%); whereas in the late stage, NC ACF (176/449, 39.2%) occurred more frequently than FC ACF (324/1,902, 17.0%). The number of ACF peaked at 15 to 20 weeks. The ratio of NC/FC ACF increased gradually during carcinogenesis. The expression of both UEA1 and PNA was higher in NC ACF than FC ACF. Lectin expression increased in the late stage compared with the early stage. CONCLUSIONS: The expression of lectin was higher in NC ACF and ACF in the late stage. Therefore, ACF with higher multiplicities in the late stage may have more malignant potential in DMH-induced colon carcinogenesis.
1,2-Dimethylhydrazine
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Aberrant Crypt Foci
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Animals
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Colon
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Dimenhydrinate
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Immunohistochemistry
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Peanut Agglutinin
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Rats
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Rats, Wistar
10.Role of Protein Kinase C in Abnormal Proliferation of Vascular Endothelial Cell induced by 1,2-Dimethylhydrazine; Analysis of Isoform.
Jin LEE ; Yong Chan BAE ; Suk Young PARK ; Jae Sul MOON ; Su Bong NAM
Journal of the Korean Society of Plastic and Reconstructive Surgeons 2007;34(1):8-12
PURPOSE: Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, have been known to take a role in signal transduction pathway of angiogenesis. The authors confirmed that PKC is the most noticeable mediator for abnormal proliferation of vascular endothelial cells through in vitro study model using the inhibitors, targeting the formation of three co-enzymes. In this study, we would investigate which isoform of PKC play an important role in abnormal angiogenesis of vascular endothelial cell. METHODS: In 96 well plates, 10(4) HUVECs(human umbilical vein endothelial cells) were evenly distributed. Two groups were established; the control group without administration of DMH(1,2-dimethylhydrazine) and the DMH group with administration of 7.5x10(-9)M DMH. RNA was extracted from vascular endothelial cell of each group and expression of the PKC isoform was analyzed by RT-PCR(reverse transcriptase-polymerase chain reaction) method. RESULTS: RT-PCR analysis showed that PKCalpha, -betaI, -betaII, -eta, -micron and -zeta were expressed in vascular endothelial cells of each group. DMH incresed the expression of PKCalpha and PKCmicron, and decreased PKCbetaI, PKCbetaII expression dominantly. CONCLUSION: Based on the result of this study, it was suggested that PKCalpha and PKCmicron may have significant role in abnormal proliferation of vascular endothelial cell.
1,2-Dimethylhydrazine*
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Cell Proliferation
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Dimenhydrinate
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Endothelial Cells*
;
Oxidoreductases
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Protein Kinase C*
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Protein Kinases*
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RNA
;
Signal Transduction
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Tyrosine
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Umbilical Veins