1.A study of gene amplification and expression of cyclin D1 in hepatocellular carcinoma.
Jian-min XU ; Jian-ming WEN ; Meng ZHANG ; Guo-li LÜ ; Li-zhi WU ; Wei-sheng WANG
Chinese Journal of Pathology 2004;33(1):26-30
OBJECTIVETo investigate gene amplification of CCND1 and expression of cyclin D1 in hepatocellular carcinoma (HCC) and to explore the possible relationship between CCND1 gene status and carcinogenesis of HCC.
METHODSDifferential PCR, RT-PCR and immunohistochemistry were used to detect gene amplification, mRNA and protein expression of cyclin D1 in 20 HCC cases respectively. The relationship between the gene amplification rate and the expression level of cyclin D1 and the histological grades of HCC was analyzed.
RESULTSCCND1 gene amplification was detected in 30% of the cases HCC. An overexpression of cyclin D1 mRNA and protein could be demonstrated in 45% and 70% cases respectively. The expression of cyclin D1 mRNA correlated with its gene amplification status (P < 0.05) and was responsible for the protein expression level (P < 0.05). There was a close relationship between the expression level of cyclin D1 protein and HCC histological grades (P < 0.05).
CONCLUSIONSCCND1 gene amplification is a common phenomenon in HCC and may be directly responsible for the cyclin D1 mRNA and protein overexpression. Cyclin D1 protein expression level is directly related to HCC histological grades. Therefore, CCND1 amplification and cyclin D1 overexpression may play an important role in development and differentiation of HCC.
Adult ; Aged ; Carcinoma, Hepatocellular ; genetics ; Cyclin D1 ; analysis ; genetics ; Female ; Gene Amplification ; Humans ; Immunohistochemistry ; Liver Neoplasms ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; RNA, Messenger ; analysis
2.Effect of estrogen on telomerase activity in human breast cancer cells.
Jinbo GAO ; Daoda CHEN ; Yuan TIAN ; Jinhui ZHANG ; Kailin CAI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(3):286-293
To investigate the effects of estrogen (E2) on telomerase activity and its mechanism in human breast cancer cells, estrogen receptor positive MCF-7 cells were treated with different concentrations of E2. Telomerase activity was measured by using TRAP-ELISA method, the cell cycle phases analyzed by using flow cytometry, and the expression of Cyclin D1 detected by using immunohistochemistry method. The results showed that telomerase activity levels were increased in MCF-7 cells treated with 10(-8) mol/L E2 during the observed period (P < 0.05), and E2 increased telomerase activity levels in a dose-dependent manner(10(-10)-10(-8) mol/L); Simultaneously, the cell cycle phases of MCF-7 cells treated with 10(-8) mol/L E2 were changed significantly: G0/G1 phase decreased from 60.52% to 50.93%. S phase increased from 29.03% to 30.83%; However, the expression of Cyclin D1 was decreased. It was concluded that estrogen can upregulate telomerase activity of MCF-7 cells, and the effect can be blocked by antiestrogen tamoxifen. Its mechanism may be closely associated with modulation of cell cycle phases.
Breast Neoplasms
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chemistry
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enzymology
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pathology
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Cell Cycle
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Cyclin D1
;
analysis
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Estrogens
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pharmacology
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Female
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Humans
;
Receptors, Estrogen
;
analysis
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Telomerase
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genetics
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metabolism
;
Tumor Cells, Cultured
3.Effect of acupoint-injection of oxymatrine on experimental hepatic carcinoma and study on the mechanism.
Ai-ling ZHOU ; Yu-juan ZHU ; Ya-e HU ; Jia-hui MAO ; Dao-rong JIANG
Chinese Acupuncture & Moxibustion 2007;27(1):48-52
OBJECTIVETo investigate the effect of acupoint injection of oxymatrine (OM) on experimental hepatocellular carcinoma and the mechanism.
METHODSThe rats of hepatocellular carcinoma induced by 2-acetoaminoflurence (2-AAF) were randomly divided into a normal control group (group N), a model group (group M), a control group of oxymatrine intraperitoneal injection (OM ip group) and a treatment group of small dose oxymatrine injection into Zusanli (OM ZSL group). At the end of 12h week, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (gamma-GT) were determined. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expressions of cyclin D1 and cyclin-dependent kinase 4 (CDK4) mRNA in hepatocellular carcinoma tissues.
RESULTSThe number of cancer nodes on the surface of liver in th Om ip group and the Om ZSL group was lower than in the group M, with the serum ALT, AST, and gamma-GT levels significantly decreased (P<0. 01), and significantly inhibited expressions of cyclin D1, CDK4 mRNA (P<0. 01).
CONCLUSIONOM ip and small dose oxymatrine injection into ZSL can treat or delay hepatocarcinogenisis of hepatocellular carcinoma induced by 2-AAF. Partial mechanism of this anti-carcinoma is protecting hepatocytes possibly through improving hepatic functions, and inhibiting excessive proliferation of liver cancer cells via inhibiting the expressions of cyclin Dl, CDK4 mRNA.
Acupuncture Points ; Alanine Transaminase ; blood ; Alkaloids ; administration & dosage ; Animals ; Aspartate Aminotransferases ; blood ; Cyclin D1 ; genetics ; Cyclin-Dependent Kinase 4 ; genetics ; Injections ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Quinolizines ; administration & dosage ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; gamma-Glutamyltransferase ; blood
4.Role of cyclinD1 and CDK4 in the carcinogenesis induced by silica.
Ke-Xia YAN ; Bing-Ci LIU ; Xiang-Lin SHI ; Bao-Rong YOU ; Ming XU
Biomedical and Environmental Sciences 2005;18(5):286-296
OBJECTIVETo study the role of cyclinD1 and CDK4 in malignant transformation of human fetal lung diploid fibroblast cell line (2BS) induced by silica.
METHODSRecombination vectors with sense and antisense pXJ41-cyclinD1 and pXJ41-CDK4 were constructed, and then transfected into the malignant transformed cells induced by silica, respectively. At the same time, pXJ41-neo was used as the control.
RESULTSDuring the progress of the malignant transformation of 2BS cells induced by silica, cyclinD1 and CDK4 were overexpressed. Antisense RNA suppressed cyclinD1 and CDK4 gene expression in the antisense pXJ41-cyclinD1 and pXJ41-CDK4 transfected cells. Antisense RNA led to cell cycle arrest, resulting in lengthened G1 phase (the percentages of cells in the G1 phase changed from 45.1% to 52.7% and 58.0% for cyclinD1 and CDK4 transfected cells, respectively), and eventually attenuated the increase of the proliferation of malignant transformed cells induced by silica. Compared with malignant transformed cells induced by silica, cells transfected with antisense pXJ41-cyclinD1 and pXJ41-CDK4 showed obviously reduced growth rates. On the 8th day, the suppression rates were 58.69 and 77.43% (the growth rate of malignant transformed cells induced by silica was 100%), doubling time changed from 21.0 h to 31.4 h and 21.0 h to 42.7 h, respectively, the growth capacities on soft agar of cells transfected by antisense pXJ41-cyclinD1 and pXJ41-CDK4 decreased obviously.
CONCLUSIONCyclinD1 and CDK4 play an important role in maintaining transformed phenotype of the cancer cells.
Carcinogens, Environmental ; toxicity ; Cell Line ; Cell Proliferation ; Cell Transformation, Neoplastic ; chemically induced ; Cyclin D1 ; genetics ; metabolism ; physiology ; Cyclin-Dependent Kinase 4 ; genetics ; metabolism ; physiology ; Humans ; Plasmids ; RNA, Antisense ; metabolism ; RNA, Messenger ; analysis ; metabolism ; Silicon Dioxide ; toxicity
5.Effect of peroxisome proliferator-activated receptor-gamma on proliferation of airway smooth muscle cells in mice with asthma.
Ming-Xiao GU ; Xuan-Cheng LIU ; Lu JIANG
Chinese Journal of Contemporary Pediatrics 2013;15(7):583-587
OBJECTIVETo investigate the effects of peroxisome proliferator-activated receptor-gamma (PPARγ) agonist rosiglitazone on the expression of cyclin D1 in lung tissue, and the proliferation of airway smooth muscle cells (ASMCs) in mice with bronchial asthma.
METHODSThirty clean BALB/c mice were randomly divided into control group (n = 10), asthma group (n = 10), and rosiglitazone treatment group (n = 10). A mouse model of asthma was established by ovalbumin (OVA) sensitization and challenge. The treatment group received rosiglitazone (5 mg/kg) by gavage 1 hour before each challenge and the control group received saline instead of OVA sensitization and challenge. Leukocytes and eosinophils in bronchoalveolar lavage fluid (BALF) were counted under a microscope. Airway structural changes were observed by hematoxylin-eosin staining. Protein and mRNA expression levels of cyclin D1 were measured by immunohistochemical staining and RT-PCR. Perimeter of the basement membrane (Pbm), total bronchial wall area (WAt), airway smooth muscle area (WAm), and number of nuclei in ASMCs (N) were determined using image analysis software, and WAt/Pbm, WAm/Pbm, and N/Pbm were calculated.
RESULTSCompared with the control group, the asthma group showed significant increases in the total number of leukocytes and percentage of eosinophils in BALF, as well as in the mRNA and protein expression of cyclin D1, but changes in these indices were significantly reduced in the rosiglitazone treatment group (P < 0.05). In addition, compared with the control group, the asthma group had significantly increased WAt/Pbm, WAm/Pbm, and N/Pbm, but rosiglitazone significantly decreased these ratios (P < 0.05).
CONCLISONSRosiglitazone may delay the process of airway remodeling by inhibiting the proliferation of ASMCs, so it can be used for preventing and treating chronic asthma.
Airway Remodeling ; Animals ; Asthma ; drug therapy ; pathology ; Bronchi ; pathology ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Proliferation ; Cyclin D1 ; analysis ; genetics ; Female ; Lung ; chemistry ; pathology ; Mice ; Mice, Inbred BALB C ; Myocytes, Smooth Muscle ; physiology ; PPAR gamma ; physiology ; RNA, Messenger ; analysis ; Thiazolidinediones ; pharmacology
6.Effects of matrine on the proliferation and apoptosis of human rhabdomyosarcoma RD cells.
Lei GUO ; Tian-Yang XUE ; Wei XU ; Ji-Zhao GAO
Chinese Journal of Contemporary Pediatrics 2012;14(10):780-784
OBJECTIVETo investigate the effects of matrine on the proliferation and apoptosis of human rhabdomyosarcoma RD cells in vitro, and to explore the mechanism of matrine inducing apoptosis of RD cells.
METHODSMTT assay was used to measure the proliferation inhibition rates of RD cells that were treated with matrine (final concentrations= 0.5, 1.0, 1.5, and 2.0 mg/mL). Flow cytometry was used to evaluate the apoptosis of RD cells treated with the four concentrations of matrine. RT-PCR was used to measure the mRNA expression of cyclin D1 and survivin in RD cells treated with 0.5, 1.0, and 1.5 mg/mL of matrine.
RESULTSThe RD cells treated with various concentrations of matrine showed significantly higher proliferation inhibition rates and apoptotic rates than those that were not treated with matrine (P<0.01), and with increased matrine concentration, the proliferation inhibition rate of RD cells increased gradually, thus exhibiting a dose dependence. The mRNA expression of cyclin D1 and survivin was seen in all RD cells, but was significantly lower in RD cells treated with matrine than in those that were not treated with matrine (P<0.01). There were significant differences in cyclin D1 mRNA level among the RD cells treated with 0.5, 1.0, and 1.5 mg/mL of matrine (P<0.05), while there was significant difference in survivin mRNA level between the RD cells treated with 0.5 and 1.5 mg/mL of matrine (P<0.05).
CONCLUSIONSMatrine can significantly inhibit proliferation and induce apoptosis of RD cells, which may be related to downregulating the mRNA expression of cyclin D1 and survivin.
Alkaloids ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin D1 ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; Quinolizines ; pharmacology ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; drug therapy ; pathology
7.Role of protein kinase C alpha and cyclin D1 in the proliferation of airway smooth muscle in asthmatic rats.
Li-fen QIAO ; Yong-jian XU ; Xian-sheng LIU ; Jun-gang XIE ; Jin WANG ; Chun-ling DU ; Jian ZHANG ; Wang NI ; Shi-xin CHEN
Chinese Medical Journal 2008;121(20):2070-2076
BACKGROUNDAirway smooth muscle (ASM) is suspected to be a determining factor in the structural change of asthma. However, the role of protein kinase C alpha (PKCalpha) and cyclin D1 involved in the dysfunction of ASM leading to asthmatic symptoms is not clear. In this study, the central role of PKCalpha and cyclin D1 in ASM proliferation in asthmatic rats was explored.
METHODSThirty-six pathogen-free male Brown Norway (BN) rats were randomly divided into 2 groups: control groups (group N1, N2 and N3) and asthmatic groups (group A1, A2, and A3). Groups A1, A2 and A3 were challenged with ovalbumin (OA) for 2 weeks, 4 weeks and 8 weeks respectively. Control animals were exposed to an aerosolized sterile phosphate buffered saline (PBS). The ASM mass and nucleus numbers were studied to estimate the degree of airway remodeling by the hematoxylin-eosin staining method. PKCalpha and cyclin D1 expression in the ASM cells was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The relation between PKCalpha and cyclin D1 was assessed by linear regression analysis. PKC agonist phorbol 12-myristate 13-acetate (PMA), PKC inhibitor Ro31-8220 and an antisense oligonucleotide against cyclin D1 (ASOND) were used to treat ASM cells (ASMCs) obtained from the 2 weeks asthmatic rats. The cyclin D1 protein expression level was detected by Western blotting.
RESULTSCompared with the control group, the PKCalpha and cyclin D1 mRNA levels were increased in the asthmatic group. Similar to RT-PCR results, immunohistochemistry analysis for PKCalpha and cyclin D1 expression revealed an increased production in ASMCs after allergen treatment for 2, 4 and 8 weeks compared with the respective control groups. No difference in expression of PKCalpha and cyclin D1 in ASM were found in the 2, 4 or 8 weeks asthmatic rats. There were significant positive correlations between PKCalpha and cyclin D1 expression, both transcriptionally (r = 0.944, P < 0.01) and translationally (r = 0.826, P < 0.01), in ASM. The content of cyclin D1 in asthmatic ASMCs increased after being stimulated by PMA, and decreased when induced by Ro31-8220. ASOND targeting for cyclin D1 lowered the expression of cyclin D1 induced by PMA.
CONCLUSIONSIncreased expression of PKCalpha and cyclin D1 in ASM along with smooth muscle structure changes might implicate PKCalpha and cyclin D1 participation in the proliferation of ASM and contribute to the pathogenesis of asthma after repeated allergen exposure in rats. The results suggested that cyclin D1 might be downstream of PKC signal transduction pathway.
Animals ; Asthma ; pathology ; Cell Proliferation ; Cyclin D1 ; genetics ; physiology ; Lung ; pathology ; Male ; Myocytes, Smooth Muscle ; pathology ; Protein Kinase C-alpha ; genetics ; physiology ; RNA, Messenger ; analysis ; Rats ; Rats, Inbred BN
8.Transfection of Lipoxin A4 receptor-like protein gene enhanced the inhibitory effect of Lipoxin A4 on human lung fibroblasts proliferation induced by connective tissue growth factor.
Chao LU ; Ji-qing CHEN ; Sheng-hua WU ; Yuan-jun WU ; Fei ZHAO ; Xiao-qin PAN ; Li FEI ; Mei GUO ; Song-ming HUANG ; Xi-rong GUO ; Rong-hua CHEN
Chinese Journal of Pediatrics 2005;43(4):288-292
OBJECTIVELipoxin A(4) is formed by the metabolism of arachidonic acid. Anti-inflammatory and anti-proliferative effect of lipoxin A(4) has been shown in many human diseases. Recently, as a novel high affinity receptor for ligand lipoxin A(4), Lipoxin A(4) receptor-like protein (LRLP) has been identified. Currently close attention is paid to the important contribution of connective tissue growth factor (CTGF) in lung fibrosis. The purpose of the study was to transfect LRLP gene into human lung fibroblasts and investigate the mechanism of its enhancing antagonistic effect of Lipoxin A(4) on human lung fibroblasts proliferation induced by connective tissue growth factor.
METHODSEukaryocytic expression vector pEGFP/LRLP which contained LRLP and green fluorescence protein fusion gene (GFP) was constructed and transfected into human lung fibroblasts (HLF). After selecting with G418, HLF/LRLP cell clone which stably expressed LRLP/GFP fusion protein was isolated and characterized by the laser scanning confocal microscope. Cultured HLF and HLF/LRLP were stimulated for 24 h with CTGF (1 microg/ml) in the presence and absence of pretreatment of Lipoxin A(4) (10.0 nmol/L) for 30 min. Inhibition of cell proliferation was determined by MTT assay. Cell cycle analysis was performed by flow cytometry. Western blot was used to detect the expression of cyclin D(1) protein. Electrophoretic mobility shift assay (EMSA) was employed to detect the DNA binding activity of STAT(3).
RESULTS(1) HLF/LRLP cell clone which stably expressed LRLP and GFP fusion protein was successfully obtained. (2) Proliferation of HLF and HLF/LRLP was induced by 1 microg/ml CTGF. Pretreatment with 10 nm Lipoxin A(4) inhibited the proliferation of HLF and HLF/LRLP. And the inhibitory rate of HLF/LRLP was significantly higher than that of HLF [(54.1 +/- 4.2)%, (21.2 +/- 3.7)%, P < 0.05]. (3) The flow cytometry analysis showed that compared with HLF, more HLF/LRLP were arrested at G(0)/G(1) phase in the presence of pretreatment of Lipoxin A(4). [(76.3 +/- 3.5)%, (60.8 +/- 2.0)%, P < 0.05]. (4) Ten nmol/L Lipoxin A(4) antagonized CTGF induced increase of cyclin D(1) protein expression in HLF and HLF/LRLP. And its antagonistic effect on HLR/LRLP was stronger than that on HLF (P < 0.05). (5) Ten nmol/L Lipoxin A(4) antagonized CTGF induced increase of STAT(3) DNA binding activity, and its antagonistic effect on HLF/LRLP was more powerful than that on HLF (P < 0.05).
CONCLUSIONSTransfection of Lipoxin A(4) receptor-like protein gene enhanced the inhibitory effect of Lipoxin A(4) on human lung fibroblasts proliferation induced by CTGF. Its mechanism might be related to regulation of cyclin D(1) protein expression and STAT(3) DNA binding activity.
Connective Tissue Growth Factor ; antagonists & inhibitors ; Cyclin D1 ; analysis ; DNA ; metabolism ; Fibroblasts ; cytology ; drug effects ; Humans ; Lipoxins ; pharmacology ; Lung ; cytology ; drug effects ; Receptors, Formyl Peptide ; genetics ; physiology ; Receptors, Lipoxin ; genetics ; physiology ; STAT3 Transcription Factor ; metabolism ; Transfection
9.Expression of CD10 in tumor-associated fibroblast of cancerized or recurrent colorectal adenomas.
Jiangjiang ZHENG ; Yin ZHU ; Changshui LI ; Yinya LI ; Qianqian NIE ; Ziling ZHU ; Hong DENG
Journal of Zhejiang University. Medical sciences 2016;45(4):335-341
To investigate the expression of CD10 in tumor-associated fibroblasts (TAF) in colorectal adenomas and its relation to cancerization and recurrence of adenoma.Tissue samples of low-grade adenoma (=50), high-grade adenoma (=50) and colorectal adenocarcinoma (=50) were collected, and tissue samples at the distal margin of corresponding colorectal lesions were taken as controls. The expression of CD10 in the stromal TAFs, and the expressions of β-catenin, Ki-67, p53 and CyclinD1 in tumor cells were detected by immunohistochemistry (Envision). The correlation of CD10 expression in stromal TAFs with the expressions of β-catenin, Ki-67, p53 and CyclinD1 in tumor cells was analyzed by Spearmen. One hundred samples of low-grade colorectal adenoma were collected, including 57 non-recurrent cases and 43 recurrent cases (16 cases of recurrent adenoma and 27 cases of recurrent adenocarcinoma); the expression of stromal TAF CD10 were determined and compared among groups.There was no TAF in normal colorectal mucosa. The expression rates of TAF CD10 in low-grade adenoma, high-grade adenoma and colorectal adenocarcinoma were 22%, 50% and 78%, respectively (all<0.05). The expression of Ki-67 and β-catenin in low-grade adenoma, high-grade adenoma, colorectal adenocarcinoma was on a rising trend (all<0.01). The expression of CyclinD1 in high-grade adenoma was higher than that in colorectal adenocarcinoma and low-grade adenoma (all>0.05). The expression of p53 in colorectal adenocarcinoma and high-grade adenoma was higher than that in low grade adenoma (all<0.01). The expression of TAF CD10 was correlated with the expression of p53, Ki-67 and β-catenin-nucleus(=0.264、0.307、0.320, all<0.01),but not correlated with CyclinD1 and β-catenin-membrane (=0.012、-0.073, all>0.05). The TAF CD10 level was significantly higher in low-grade adenoma with recurrence than that in those without recurrence (<0.05).The expression of CD10 in recurrent colorectal adenocarcinoma was higher than that in recurrent adenoma (<0.05).The expression of TAF CD10 is increased gradually in the process of adenoma-cancer, indicating that it may play an important role in the canceration of adenoma. Adenomas with high expression of CD10 TAF are likely to be recurrent and cancerized, and detection of TAF CD10 combined with p53, Ki-67 and β-catenin may be of value in predicting canceration or recurrence of colorectal adenoma.
Adenocarcinoma
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chemistry
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genetics
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Adenoma
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chemistry
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genetics
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Biomarkers, Tumor
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analysis
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Cancer-Associated Fibroblasts
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chemistry
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Carcinogenesis
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chemistry
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Colorectal Neoplasms
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chemistry
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genetics
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Cyclin D1
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analysis
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Disease Progression
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Humans
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Immunohistochemistry
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Ki-67 Antigen
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analysis
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Neoplasm Grading
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Neoplasm Recurrence, Local
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chemistry
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Neprilysin
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analysis
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Predictive Value of Tests
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Tumor Suppressor Protein p53
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analysis
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beta Catenin
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analysis
10.Comparison of Her-2, EGFR and Cyclin D1 in Primary Breast Cancer and Paired Metastatic Lymph Nodes: An Immunohistochemical and Chromogenic In Situ Hybridization Study.
Eun Yoon CHO ; Jae Joon HAN ; Yoon La CHOI ; Kyoung Mee KIM ; Young Lyun OH
Journal of Korean Medical Science 2008;23(6):1053-1061
The significant advance in the development of molecular-targeting drugs has made an evaluation of Her-2, EGFR, and cyclin D1 an important clinical issue in breast cancer patients. This study compared the Her-2, EGFR, and cyclin D1 status of primary tumors as well as their matching lymph node metastases using immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) in 73 breast cancer patients. Her-2, EGFR, and cyclin D1 protein showed a concordance between the primary lesion and the metastatic regional lymph nodes in 82%, 90%, and 63%, respectively. CISH also revealed 92%, 93%, and 85% concordance in the gene amplification status of Her-2, EGFR, and cyclin D1, showing a reasonable agreement between primary tumors and metastatic regional lymph nodes. Although a statistically significant agreement was found in Her-2 expression, a relatively high discordance rate (18%) raises a little concern. Our findings suggest that the Her-2 status can be reliably assessed on primary tumor but a possible difference can be found in Her-2, EGFR, and cyclin D1 status between the primary and the metastatic sites and this possibility should be concerned in patients considering molecular targeted therapy or patients with progress of disease.
Adult
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Aged
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Breast Neoplasms/genetics/*metabolism/pathology
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Chromogenic Compounds
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Cyclin D1/*analysis/genetics
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Female
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Humans
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Immunohistochemistry
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In Situ Hybridization
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Lymph Nodes/*metabolism/pathology
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Lymphatic Metastasis
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Male
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Middle Aged
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Neoplasm Recurrence, Local/genetics/metabolism
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Receptor, Epidermal Growth Factor/*analysis/genetics
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Receptor, erbB-2/*analysis/genetics
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Survival Analysis