1.Immunohistochemical demonstration of cyclins A, B, D1, D3 and E in hepatocellular carcinomas using tissue microarrays.
Ming-hua ZHU ; Can-rong NI ; Zhi ZHU ; Fang-mei LI ; Shun-min ZHANG
Chinese Journal of Pathology 2003;32(5):440-443
<b>OBJECTIVEb>To investigate the expression of five kinds of cyclins in hepatocellular carcinoma (HCC) and their association with degree of tumor differentiation, metastasis and infection of hepatitis B virus (HBV).
<b>METHODSb>The HCC tissue microarrays were composed of those from 273 cases of HCC tissues, 144 surrounding-tumor liver tissues and 10 normal liver tissues obtained from autopsy. The diameter of each specimens on tissue microarrays was 2.0 mm. Immunohistochemistry was used to detect the expression of cyclin A, cyclin B, cyclin D1, cyclin D3 and cyclin E on HCC tissue microarrays. The association of the expression of these cyclins with the infection rate of HBV was also analyzed.
<b>RESULTSb>Three paraffin-embedded HCC tissue microarrays were successfully constructed, including 136, 143 and 148 tissue spots, respectively. The positive rates of cyclins in 273 cases of HCC were cyclin A 52.7%, cyclin B 45.4%, cyclin D1 35.9%, cyclin D3 44.3% and cyclin E 23.1%; while the figures in 144 surrounding-tumor tissues were 8.3%, 5.6%, 4.9%, 6.3% and 1.4%, respectively. In 10 normal liver tissues these cyclins exhibited negative staining, with the exception that cyclin D1 was positive in one case of normal liver tissue. The positive rate of cyclins in HCC were significant higher than those in surrounding-tumor liver tissues (P < 0.01), in HCC tissues with histological grade II and III, the cyclins expression were stronger than that in grade I (P < 0.05). The positive rates of cyclins, except cyclin A in HCC with portal vein invasion were higher than those without portal vein invasion (P < 0.01). Infection of HBV did not have significant relationship with the expression of cyclins (P > 0.05).
<b>CONCLUSIONb>Cyclins in different cell cycles overexpressed at varied levels in hepatocellular carcinoma, and the increased expression of cyclins may shorten the tumor cell cycle phase, accelerate cell proliferation, and have a close relationship with HCC aggressiveness.
Carcinoma, Hepatocellular ; chemistry ; Cyclin A ; analysis ; Cyclin B ; analysis ; Cyclin D1 ; analysis ; Cyclin D3 ; Cyclin E ; analysis ; Cyclins ; analysis ; Hepatitis B ; metabolism ; Humans ; Immunohistochemistry ; Liver Neoplasms ; chemistry
2.Overexpressions of Cyclin B1, cdc2, p16 and p53 in Human Breast Cancer: The Clinicopathologic Correlations and Prognostic Implications.
Seoung Wan CHAE ; Jin Hee SOHN ; Dong Hoon KIM ; Yoon Jung CHOI ; Yong Lai PARK ; Kyungeun KIM ; Young Hye CHO ; Jung Soo PYO ; Jun Ho KIM
Yonsei Medical Journal 2011;52(3):445-453
PURPOSE: The molecular mechanisms that are responsible for the initiation and progression of breast cancers are largely unknown. This study was to analyze the cyclin B1, cdc2, p53 and p16 tumor suppressor genes in human breast cancer. MATERIALS AND METHODS: To investigate the role of cyclin B1, cdc2, p53 and p16 in the pathogenesis and progression of breast carcinomas, 98 cases of breast cancers were examined by immunohistochemical method. The correlations of cyclin B1, cdc2, p53 and p16 expression with various clinico-pathologic findings were analysed. RESULTS: In the normal breast tissues, cyclin B1, cdc2 and p16 were weakly expressed, while p53 was not expressed. On the other hand, cyclin B1, cdc2, p53 and p16 were overexpressed in breast cancer, showing correlation between the expression of cyclin B1 and cdc2 and breast cancers (p=0.00). The overexpressions of cdc2 and p16 were correlated with an infiltrative tumor border pattern and this was statistically significant (p<0.05). In addition, the overexpression of cdc2 was correlated with histologic high grade carcinomas (p=0.00). CONCLUSION: Cyclin B1 and cdc2 appeared to be involved in the genesis or progression of breast cancers. In addition, the overexpressions of p16 and p53 may play important roles in more aggressive tumor and the overexpression of cdc2 is associated with progression of tumor to a higher grade of breast carcinomas. The deranged overexpressions of cyclin B1, cdc2, p16 and p53 may play an important role in human breast carcinogenesis.
Adult
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Aged
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Breast Neoplasms/*genetics/metabolism/pathology
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Cyclin B/*genetics/metabolism
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Cyclin B1/*genetics/metabolism
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Cyclin-Dependent Kinase Inhibitor p16/*genetics/metabolism
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Female
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Gene Expression Regulation, Neoplastic
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Humans
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Immunohistochemistry
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Middle Aged
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Tumor Suppressor Protein p53/*genetics/metabolism
3.The clinical significance of cyclin B1 expression in adult acute leukemia patients.
Wei-dong MA ; Shi-rong XU ; Xiao-nan GUO ; Jin-song JIA ; Fang XUE
Chinese Journal of Hematology 2003;24(10):523-526
<b>OBJECTIVEb>To investigate the clinical significance of cyclin B1 expression in adult acute leukemia (AL) patients.
<b>METHODSb>The expression of cyclin B1 and p21 and their cell cycle distribution were measured by flow cytometry in 85 adult patients with de novo AL, 10 continuous complete remission (CCR) AL and 17 normal controls (NC). The mRNAs of cyclin B1, p21 cip1 and proliferation cell nuclear antigen (PCNA) in patients and NCs were measured with semi-quantity reverse transcription polymerase chain reaction (RT-PCR).
<b>RESULTSb>Cyclin B1 protein expression in de novo AL patients was significantly higher than that in NC (P < 0.001). It was higher in relapsed patients than in NC (P < 0.05) but was lower than in de novo AL (P < 0.01). There was no difference between the remission cases and NC (P = 0.21), and between CCR patients and NC (P > 0.05). The cyclin B1 overexpression ratio was higher than that of NC. A negative correlation between the expression levels of cyclin B1 and P21 was observed (r = -0.266, P < 0.05). The cyclin B1 protein expression level was positively correlated with its mRNA level. The expression of cyclin B1 was positively correlated with proliferation index (PI) levels, and with PCNA levels (rPI = 0.7314, rPCNA = 0.7152). Remission rate was higher in high cyclin B1 expression patients than in normal cyclin B1 expression patients (P < 0.01), so did the relapse rate (P < 0.01). Patients with higher cyclin B1 expression had higher survival rate.
<b>CONCLUSIONb>Cyclin B1 was overexpressed and abnormally distributed in cell cycle phases in de novo AL patients. Overexpression of cyclin B1 might be a favorable prognostic factor for patients with AL.
Adolescent ; Adult ; Cell Division ; Cyclin B ; analysis ; Cyclin B1 ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; analysis ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; mortality ; therapy ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; mortality ; therapy ; Prognosis ; Recurrence ; Survival Rate
4.Effect of lovastatin on the expression of IkappaBalpha and cell-cycle regulating protein in MCF-7 cells.
Na WEI ; Man-tian MI ; Qian-yong ZHANG ; Zhi-xiang YANG
Chinese Journal of Oncology 2003;25(4):332-334
<b>OBJECTIVEb>To study the effect of lovastatin on the expression of IkappaBalpha and cell-cycle regulating proteins in MCF-7 cells.
<b>METHODSb>MCF-7 cells were treated with 4, 8 and 16 micro mol/L lovastatin for 48 - 72 h. The distribution of cell cycles was assayed by flow cytometry (FCM). The protein expression of IkappaBalpha, CDK4, p16, pRb in cytoplasm and IkappaBalpha in the nucleus were detected by Western blot.
<b>RESULTSb>Lovastatin could arrest cellcycle in the G(0)/G(1) phase in a dose- and time-dependent manner, obviously lowering the expression of IkappaBalpha, CDK4 and pRb protein level in the cytoplasm and increasing IkappaBalpha in the nucleus, but not on p16 protein level.
<b>CONCLUSIONb>Lovastatin can induce the arrest of cell cycle in G(0)/G(1) phase by affecting the expression of IkappaBalpha and cell-cycle regulating protein in MCF-7 cells.
Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Cyclin-Dependent Kinases ; metabolism ; Female ; Humans ; I-kappa B Proteins ; metabolism ; Lovastatin ; pharmacology ; NF-KappaB Inhibitor alpha ; NF-kappa B ; antagonists & inhibitors ; Proto-Oncogene Proteins ; metabolism ; Retinoblastoma Protein ; metabolism
6.Influence of aerosols on the expression of cyclin B1, cyclin C and proliferating cell nuclear antigen in wound tissue healing of burned rat models.
Tao ZHANG ; Tian-zeng LI ; Ying-bin XU ; Shao-hai QI ; Hui-zhen LIANG ; Ju-lin XIE ; Xiao-dong CHEN ; Shu-sen ZHENG
Chinese Journal of Surgery 2005;43(19):1280-1283
<b>OBJECTIVEb>To investigate the influence of aerosols on the expression of cyclin B(1), cyclin C and proliferating cell nuclear antigen (PCNA) in wound tissue healing of burned rat models.
<b>METHODSb>Sprague Dawley (SD) rats were inflicted as the deep partial thickness burn models. Rats were randomly divided into experimental group and control group. The experimental group were treated with aerosols. Samples were collected in 1 approximately 10 postburn days. Immunohistochemistry and image analysis methods were conducted to examine the expression of cyclin B(1), cyclin C and PCNA in both experimental and control groups.
<b>RESULTSb>The expression of cyclin C in experimental group was detected in nucleus of skin basal cell on the second postburn day, increased evidently at the fifth days and sustained at high expression level up to the tenth days after treatment. The expression of cyclin C in experimental group was significantly higher than control group (P < 0.05). The expression of PCNA was first observed in skin basal cell nucleus and hair follicle cell nucleus in both experimental and control group on the third postburn day. The expression of PCNA increased evidently at the fifth days in experimental after treatment and that increased evidently at the seventh days in control group, which showed there were lots of active proliferation cell. And the difference of the expression of PCNA between experimental and control group was significant (P < 0.01). The expression of cyclin B(1) was detected in nucleus and cytoplasm of skin basal cell in both groups on the third postburn day, and no difference between the experimental and control group (P > 0.05).
<b>CONCLUSIONSb>Aerosols can up-regulate the expression of cyclin C and PCNA in skin basal cell nucleus. Therefore the aerosols can accelerate wound tissue healing.
Aerosols ; Animals ; Burns ; metabolism ; therapy ; Cyclin B ; biosynthesis ; Cyclin B1 ; Cyclin C ; Cyclins ; biosynthesis ; Disease Models, Animal ; Electric Stimulation Therapy ; methods ; Female ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Wound Healing ; physiology
7.Clinical Significance of Detecting CyclinD1 and BCL-2 in Patients with B-Cell Lymphoma by Using Flow Cytometry.
Yan-Xiang CUI ; Xin-Hua WANG ; Xun CHEN
Journal of Experimental Hematology 2016;24(1):98-101
<b>OBJECTIVEb>To explore the feasibility and value of detecting CyclinD1 and BCL-2 in patients with B-cell lymphoma by using flow cytometry.
<b>METHODSb>Fifty-three patients with lymphoma were selected, and 50 healthy persons in the same period were selected as control. The expression levels of CyclinD1 and BCL-2 in patients with various subtypes of lymphoma were detected by using flow cytometry (FCM).
<b>RESULTSb>When the dilution time was 1 min and the dilution proportion was 1:20, the cell morphology was the best complete, at the 4 min the cell morphology was best status. The mean fluorescence intensity of CyclinD1 and BCL-2 between persons of control group and patients with B-cell lymphoma showed significant difference, the CyclinD1 level (1.824 ± 0.315) and BCL-2 levels (4.257 ± 0.528) of patients with B-cell lymphoma were obviously higher than the CyclinD1 level (0.634 ± 0.153) and BCL-2 level (1.926 ± 0.328) of persons in control group, the CyclinD1 and BCL-2 expression levels of patients with HL were significantly lower than CyclinD1 and BCL-2 levels of patients with NHL (P < 0.01). After treatment, the expression levels of CyclinD1 and BCL-2 in patients with B lymphoma were significantly lower than these befor treatment.
<b>CONCLUSIONb>Using the method of flow cytometry for detecting CyclinD1 and BCL-2 expression levels in lymphoma cells of patients is feasible, and it can be applied clinically to evaluate the treatment efficacy.
Case-Control Studies ; Cyclin D1 ; metabolism ; Flow Cytometry ; Humans ; Lymphoma, B-Cell ; diagnosis ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism
8.Tetrandrine: a potent abrogator of G2 checkpoint function in tumor cells and its mechanism.
Xin-Chen SUN ; Hong-Yan CHENG ; Yu-Xia DENG ; Rong-Guang SHAO ; Jun MA
Biomedical and Environmental Sciences 2007;20(6):495-501
<b>OBJECTIVEb>To assess the ability of tetrandrine (Tet) to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcinoma p53-mutant HT-29 as well as in C26 colorectal carcinoma-bearing BALB/c mice.
<b>METHODSb>MCF-7/ADR, HT-29 and MCF-7 cells were exposed to irradiation in the absence or presence of tetrandrine. The effect of Tet on the cytotoxicity of X-irradiation in these three cells was determined and the effect of tetrandrine on cell cycle arrest induced by irradiation in its absence or presence was studied by flow cytometry. Moreover, mitotic index measurement determined mitosis of cells to enter mitosis. Western blotting was employed to detect cyclin B 1 and Cdc2 proteins in extracts from irradiated or non-irradiated cells of MCF-7/ADR, HT-29 and MCF-7 treated with tetrandrine at various concentrations. Tumor growth delay assay was conducted to determine the radio-sensitization of tetrandrine in vivo.
<b>RESULTSb>Clonogenic assay showed that tetrandrine markedly enhanced the lethal effect of X-rays on p53-mutant MCF-7/ADR and HT-29 cells and the sensitization enhancement ratio (SER) of tetrandrine was 1.51 and 1.63, but its SER was only 1.1 in p53-wt MCF-7 cells. Irradiated p53-mutant MCF-7/ADR and HT-29 cells were only arrested in G2/M phase while MCF-7 cells were arrested in G1 and G2/M phases. Radiation-induced G2 phase arrests were abrogated by tetrandrine in a concentration-dependent manner in MCF-7/ADR and HT-29 cells, whereas redistribution within MCF-7 cell cycle changed slightly. The proportion of cells in M phase increased from 1.3% to 14.7% in MCF-7/ADR cells, and from 1.5% to 13.2% in HT-29 cells, but 2.4% to 7.1% in MCF-7 cells. Furthermore, the levels of cyclin B 1 and Cdc2 expression decreased after X-irradiation in MCF-7/ADR and HT-29 cells, and the mitotic index was also lower. Tet could reverse the decrease and induce the irradiated cells to enter mitosis (M phase). Endosomatic experiment showed that tetrandrine caused tumor growth delay in irradiated mice.
<b>CONCLUSIONb>Tetrandrine boosts the cell killing activity of irradiation both in vitro and in vivo. Tetrandrine is a potent abrogator for G2 checkpoint control and can sensitize the cells to radiation.
Animals ; Benzylisoquinolines ; pharmacology ; CDC2-CDC28 Kinases ; metabolism ; Cell Line, Tumor ; Cyclin B ; metabolism ; Cyclin B1 ; Drug Screening Assays, Antitumor ; G2 Phase ; drug effects ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Radiation Tolerance
9.Analysis of the genetic interactions between Cyclin A1, Atm and p53 during spermatogenesis.
Nicole BAUMER ; Marie-Luise SANDSTEDE ; Sven DIEDERICHS ; Gabriele KOHLER ; Carol READHEAD ; Ping JI ; Feng ZHANG ; Etmar BULK ; Jorg GROMOLL ; Wolfgang E BERDEL ; Hubert SERVE ; Carsten MULLER-TIDOW
Asian Journal of Andrology 2007;9(6):739-750
<b>AIMb>To analyze the functional interactions of Cyclin with p53 and Atm in spermatogenesis and DNA double-strand break repair.
<b>METHODSb>Two lines of double knockout mice were generated. Spermatogenesis and double strand break repair mechanisms were analyzed in Cyclin A1 (Ccna1); p53- and Ccna1; Atm-double knockout mice.
<b>RESULTSb>The block in spermatogenesis observed in Cyclin A1-/- (Ccna1-/-) testes at the mid-diplotene stage is associated with polynucleated giant cells. We found that Ccna1-deficient testes and especially the giant cells accumulate unrepaired DNA double-strand breaks, as detected by immunohistochemistry for phosphorylated H2AX. In addition, the giant cells escape from apoptosis. The development of giant cells occurred in meiotic prophase I, because testes lacking ATM, which are known to develop spermatogenic arrest earlier than prophase I, do not develop giant cells in the absence of cyclin A1. Cyclin A1 interacted with p53 and phosphorylated p53 in complex with CDK2. Interestingly, p53-deficiency significantly increased the number of giant cells in Ccna1-deficient testes. Gene expression analyses of a panel of DNA repair genes in the mutant testes revealed that none of the genes examined were consistently misregulated in the absence of cyclin A1.
<b>CONCLUSIONb>Ccna1-deficiency in spermatogenesis is associated with defects in DNA double-strand break repair, which is enhanced by loss of p53.
Animals ; Ataxia Telangiectasia Mutated Proteins ; Cell Cycle ; genetics ; physiology ; Cell Cycle Proteins ; genetics ; metabolism ; Cyclin A ; genetics ; metabolism ; Cyclin A1 ; Cyclin B ; Cyclin B2 ; DNA ; genetics ; DNA Repair ; genetics ; physiology ; DNA-Binding Proteins ; genetics ; metabolism ; Gene Expression Regulation ; genetics ; physiology ; Male ; Mice ; Mice, Knockout ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Spermatogenesis ; genetics ; physiology ; Testis ; cytology ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Tumor Suppressor Proteins ; genetics ; metabolism
10.Crystal structure of human Gadd45γ corrected reveals an active dimer.
Wenzheng ZHANG ; Sheng FU ; Xuefeng LIU ; Xuelian ZHAO ; Wenchi ZHANG ; Wei PENG ; Congying WU ; Yuanyuan LI ; Xuemei LI ; Mark BARTLAM ; Zong-Hao ZENG ; Qimin ZHAN ; Zihe RAO
Protein & Cell 2011;2(10):814-826
The human Gadd45 protein family plays critical roles in DNA repair, negative growth control, genomic stability, cell cycle checkpoints and apoptosis. Here we report the crystal structure of human Gadd45γ [corrected], revealing a unique dimer formed via a bundle of four parallel helices, involving the most conserved residues among the Gadd45 isoforms. Mutational analysis of human Gadd45γ [corrected] identified a conserved, highly acidic patch in the central region of the dimer for interaction with the proliferating cell nuclear antigen (PCNA), p21 and cdc2, suggesting that the parallel dimer is the active form for the interaction. Cellular assays indicate that: (1) dimerization of Gadd45γ [corrected] is necessary for apoptosis as well as growth inhibition, and that cell growth inhibition is caused by both cell cycle arrest and apoptosis; (2) a conserved and highly acidic patch on the dimer surface, including the important residues Glu87 and Asp89, is a putative interface for binding proteins related to the cell cycle, DNA repair and apoptosis. These results reveal the mechanism of self-association by Gadd45 proteins and the importance of this self-association for their biological function.
Amino Acid Motifs
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Animals
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Apoptosis
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radiation effects
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CDC2 Protein Kinase
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Cell Cycle
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Cell Survival
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Crystallography, X-Ray
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Cyclin B
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metabolism
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Cyclin-Dependent Kinase Inhibitor p21
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metabolism
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Cyclin-Dependent Kinases
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HeLa Cells
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Humans
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Intracellular Signaling Peptides and Proteins
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chemistry
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genetics
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metabolism
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Mice
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Mutagenesis, Site-Directed
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Mutation, Missense
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Proliferating Cell Nuclear Antigen
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metabolism
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Protein Binding
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Protein Interaction Domains and Motifs
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Protein Multimerization
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Protein Structure, Quaternary
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Ultraviolet Rays