<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

OBJECTIVES: The molecular mechanisms that regulate cardiomyocyte cell cycle and terminal differentiation in humans remain largely unknown. To determine which cyclins, cyclin dependent kinases (CDKs) and cyclin kinase inhibitors (CKIs) are important for cardiomyocyte proliferation, we have examined protein levels of cyclins, CDKs and CKIs during normal atrial development in humans. METHODS: Atrial tissues were obtained in the fetus from inevitable abortion and in the adult during surgery. Cyclin and CDK proteins were determined by Western blot analysis. CDK activities were determined by phosphorylation amount using specific substrate. RESULTS: Most cyclins and CDKs were high during the fetal period and their levels decreased at different rates during the adult period. While the protein levels of cyclin D1, cyclin D3, CDK4, CDK6 and CDK2 were still detectable in adult atria, the protein levels of cyclin E, cyclin A, cyclin B, cdc2 and PCNA were not detectable. Interestingly, p27KIP1 protein increased markedly in the adult period, while p21CIP1 protein in atria was detectable only in the fetal period. While the activities of CDK6, CDK2 and cdc2 decreased markedly, the activity of CDK4 did not change from the fetal period to the adult period. CONCLUSION: These findings indicate that marked reduction of protein levels and activities of cyclins and CDKs, and marked induction of p27KIP1 in atria, are associated with the withdrawal of cardiac cell cycle in adult humans.
Adult ; Age Factors ; Animal ; Blotting, Western ; Cell Cycle ; Cells, Cultured ; Comparative Study ; Cyclin A/analysis ; Cyclin B/analysis ; Cyclin D1/analysis ; Cyclin E/analysis ; Cyclin-Dependent Kinases/analysis* ; Cyclins/analysis* ; Female ; Fetal Development ; Heart Atrium/growth & development* ; Heart Atrium/embryology ; Heart Atrium/cytology* ; Heart Atrium/chemistry ; Human ; Male ; Middle Age ; Myocardium/chemistry* ; Rats ; Rats, Sprague-Dawley ; Substances: Cyclin D1 ; Substances: Cyclins ; Substances: Cyclin-Dependent Kinases ; Substances: Cyclin E ; Substances: Cyclin B ; Substances: Cyclin A

<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

<b>OBJECTIVEb>To investigate the content and activity of M-phase promoting factor (MPF) in pleomorphic adenoma, mucoepidermoid carcinoma, buccal carcinoma and normal tissue, in order to evaluate the role of MPF in the development of tumor and the relationship between MPF and malignant degree.

<b>METHODSb>The content and activity of MPF were assessed by immunobloting and Gollicano method.

<b>RESULTSb>The cdc2 and cyclinB (two subunits of MPF) were found both in normal and tumor tissues, and their content in tumor was higher than normal tissues. Buccal carcinoma was 64% higher than normal tissues. The activity of MPF in carcinoma was higher than normal tissue and had positive relation with the malignant extent.

<b>CONCLUSIONSb>The content and activity of MPF in tumor are higher than normal tissue. PKC can activate MPF. These results show PKC may promote tumor proliferation by activating MPF and also, the activity of MPF has some relation with malignant extent.

CDC2 Protein Kinase ; analysis ; Cyclin B ; analysis ; Humans ; Immunoblotting ; Maturation-Promoting Factor ; analysis ; Mouth ; chemistry ; Mouth Neoplasms ; chemistry ; Protein Kinase C ; physiology

In order to investigate the role and the mechanism of protein tyrosine phosphatase (PTPase) signaling pathway in the regulation of proliferation, cell cycle and apoptosis in lymphoma cells, the effects of sodium orthovanadate, Na(3)VO(4), a specific PTPase inhibitor, were explored on Raji lymphoblast-like cell line by MTT assay and CFU-Raji culture, morphologic observation, DNA gel electrophoresis, FCM and RT-PCR. Results showed that MTT assay and CFU-Raji culture demonstrated that sodium or thovanadate inhibited the growth of Raji cells in a concentration-dependent fashion; morphologic observations showed that Raji cells exhibited cytoplasm shrinkage, cytoplasm membrane blebbing, nuclear fragmentation and chromatin condensation forming crescents along nuclear membrane characteristic of apoptosis in the presence of Na(3)VO(4); DNA gel electrophoresis revealed typical DNA ladder reminiscent of DNA cleavage at internucleosomal sites in Na(3)VO(4) treated cells; FCM and RT-PCR indicated that Na(3)VO(4) intervention increased the fraction of annexin V(+) PI(-) cells, reduced the value of mitochondrial transmembrane potential, induced G(2)/M arrest and down-regulated the expression of Bcl-2 and cyclin B1 at both mRNA and protein level in a concentration-dependent manner. It was concluded that PTPase pathway might be implicated in the regulation of cell proliferation, cell cycle and apoptosis, and PTPase specific inhibitor Na(3)VO(4) could induce Raji cell growth inhibition, G(2)/M arrest and apoptosis via down-regulation of Bcl-2 and cyclin B1, and reduction of mitochondrial transmembrane potential.
Apoptosis ; drug effects ; Cell Division ; drug effects ; Cyclin B ; analysis ; Cyclin B1 ; Enzyme Inhibitors ; pharmacology ; Humans ; Leukocyte Common Antigens ; analysis ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; physiology ; Protein Tyrosine Phosphatases ; antagonists & inhibitors ; Vanadates ; pharmacology

<b>OBJECTIVEb>To explore the prognostic factors of primary nodal diffuse large B-cell lymphomas (N-DLBCL).

<b>METHODSb>According to the 2001 WHO classification of tumors of hematopoietic and lymphoid tissue, 51 cases of primary N-DLBCL were collected for clinical data analysis and immunohistochemical assay. Antibodies used for study were anti-CD20, CD79alpha, CD45RO, CD3, Bcl-2, Ki-67, CD30, CD15, kappa, lambda, Cyclin D1, TdT, GFAP, CK, MPO. The survival data was analyzed.

<b>RESULTSb>Of the 51 cases of N-DLBCLs, 40 were reclassified as centroblastic, 3 B-immunoblastic, 1 T-cell/histiocytes rich, 2 B-cell anaplastic large cell, 1 plasmablastic, and 4 unclassified. Expression of Bcl-2 oncoprotein was observed in 24 cases (47.1%). The median Ki-67 index was 50.0% and the index more than 40% was found in 35 cases (68.6%). Survival analysis of 35 cases had follow up data showed that the 2 year and 5-year overall survival (OS) rates were 48.54% and 35.30%, respectively. The 5-year OS rates patients with International Prognosis Index (IPI) >/= 3 was lower than that with IPI < 3 (P < 0.01). The 5-year OS rates for patients with B symptoms was lower than that without B symptoms (P < 0.05). The 5-year OS rates for patients with Ki-67 index more than 40% was lower than that with less than 40% (P < 0.05). The expression of Bcl-2 oncoprotein was uncorrelated to prognosis (P > 0.05).

<b>CONCLUSIONb>IPI, B symptoms and Ki-67 index are the prognostic factors for patients with N-DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; analysis ; Child ; Child, Preschool ; Cyclin D1 ; analysis ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Ki-1 Antigen ; analysis ; Ki-67 Antigen ; analysis ; Leukocyte Common Antigens ; analysis ; Lewis X Antigen ; analysis ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Survival Analysis ; Young Adult

<b>OBJECTIVEb>To study the enhancing effect of isoflavonoid genistein in irradiation (IR) on prostate DU145 cancer cells.

<b>METHODSb>Prostate cancer cell line DU145 was used in this experiment. Clonogenic assay was applied to compare the survival fractions of DU145 cells after treatments with genistein alone and/or graded IR. DNA electrophoresis and TUNEL method were applied to detect cell apoptosis. Cell cycle was observed using flow cytometry and related protein expressions by immunoblotting.

<b>RESULTb>Clonogenic assay demonstrated that genistein, even at low to medium concentrations, enhanced the radiosensitivity of DU145 cells. After treatments with IR and/or genistein for 24 h, apoptosis was mainly seen with genistein at high concentration and was minimally dependent on IR. Apoptosis also occurred after treatments for 72 h with lower concentrations of genistein, especially when combined with IR. While IR or genistein led to a G2/M cell cycle arrest, combination of them could further increase DU145 cells at G2/M phase. This G2/M arrest was largely maintained at 72 h, and accompanied by increasing apoptosis and hyperdiploid cell populations. Cell-cycle related protein analysis disclosed biphasic changes in cyclin B1, less markedly increased cdc-2 and stably elevated p21(cip1) levels with increasing genistein concentrations.

<b>CONCLUSIONb>Genistein could enhance the radiosensitivity of DU145 prostate cancer cells. The mechanisms might be involved in the increased apoptosis, prolonged cell cycle arrest and impaired damage repair induced by the combined treatment.

Apoptosis ; drug effects ; radiation effects ; CDC2 Protein Kinase ; analysis ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Cyclin B ; analysis ; Cyclin B1 ; G2 Phase ; drug effects ; radiation effects ; Genistein ; pharmacology ; Humans ; Male ; Prostatic Neoplasms ; pathology ; radiotherapy ; Radiation-Sensitizing Agents ; pharmacology ; S Phase ; drug effects ; radiation effects

Deranged expression of cell cycle modulators has been reported to contribute to the development and progression of hepatocellular carcinoma (HCC). However, their expression patterns remain poorly understood in hepatitis B virus (HBV)-related HCC, which constitutes about 65-70% of HCC in Korea. The aims of this study were to evaluate the expressions of G1-S modulators in HBV-related HCCs and dysplastic nodules (DNs), and to correlate with the histopathologic features of HCCs. Immunohistochemical expressions of cyclin D1, cyclin E, p53, p27, p21, p16, Rb, and PCNA proteins were investigated in 80 HCCs and 22 DNs. Cyclin D1 overexpression showed positive relationships with advanced tumor stage, poor differentiation, larger tumor size, microvascular invasion, intrahepatic meta-stasis, no tumor capsule formation, infiltrative growth, aberrant p53 expression, and high PCNA labeling index (LI) of HCC (p<0.05). Aberrant p53 expression showed positive relationship with poor differentiation of HCC (p<0.01). Expression of cyclin D1 or p53 was not observed in DNs. The p27 LI and p16 LI were lower in HCCs with intrahepatic metastasis (p<0.05). Cyclin D1 overexpression and aberrant p53 expression could be associated with the progression of HBV-related HCC, and might have a less crucial role in the DN-HCC sequence. In addition, elevated expression of p27 and p16 proteins might have inhibitory action to the intrahepatic metastasis of HBV-related HCC.
Adult ; Aged ; Carcinoma, Hepatocellular/chemistry/etiology/*pathology ; Cyclin D1/*analysis ; Female ; G1 Phase ; Hepatitis B/*complications ; Human ; Immunohistochemistry ; Liver Neoplasms/chemistry/etiology/*pathology ; Male ; Microfilament Proteins/analysis ; Middle Age ; Precancerous Conditions/*virology ; Proliferating Cell Nuclear Antigen/analysis ; Protein p16/analysis ; Protein p53/*analysis ; Retinoblastoma Protein/analysis ; S Phase

A case with mantle cell lymphoma, blastic variant, involving the peripheral blood in a 78- year-old female was described. The circulating blast-like cells suggested the possibility of acute leukemia. The WBC count was 28.0x109/ L with the absolute lymphoctyosis, 14.0x109/L. The peripheral blood smears showed a medium to large-sized lymphocytes with blast- like chromatin and conspicuous nucleoli. Atypical cells showing cleaved or indented nucleus were frequently observed. By flow cytometric analysis, immunophenotype of the circulating leukemic cells expressed B-cell associated antigens (CD 19, CD20), coexpressed CD5, lacked CD23, and expressed monoclonal surface immunoglobulin. Bone marrow aspiration and bone core biopsy confirmed diffuse infiltration of lymphoma cells. Immunohistochemicalstain for cyclin D1 (PRAD1/Bcl-1) protein was strongly reactive in the nucleus and cytoplasm of the infiltrative lymphoma cells in the bone marrow. Cytogenetic analysis of the bone marrow aspirates showed the complex abnormalities. Heterotransplantations of the blastic mantle cell leukemic cells, 4x104/microliter, into the peritoneum of the severe combined immunodeficiency (SCID) mouse showed tumor formation and infiltration of malignant lymphoid cells into peritoneum, liver, bone marrow, diaphragm, spinal cord and testes. The histopathology, immunophenotype and cyclin D1 protein expression of xenotransplanted tumor showed identical findings that of the original peripheral blood.
Animals ; B-Lymphocytes ; Biopsy ; Bone Marrow ; Chromatin ; Cyclin D1 ; Cyclins ; Cytogenetic Analysis ; Cytoplasm ; Diaphragm ; Female ; Humans ; Immunoglobulins ; Leukemia ; Liver ; Lymphocytes ; Lymphoma ; Lymphoma, Mantle-Cell* ; Mice ; Peritoneum ; Severe Combined Immunodeficiency ; Spinal Cord ; Testis

<b>OBJECTIVEb>To detect the gene expression profile in gastric cancer cell cycle and explain the mechanism of gastric cancer cell proliferation by a genomic study.

<b>METHODSb>Gastric cancer cells MKN45 were synchronized at G2/M and G1/S point by nocodazole-thymidine and double thymidine methods. The synchronizing degree of cells was monitored by flow cytometry. The gene expression profiles at G2/M point, M/G1 transition, G1 early phase, G1 late phase, G1/S point, S early phase, S late phase, G2 early phase and G2 late phase in MKN45 cell cycling were examined using cDNA microarray chips. Hierarchy analysis was conducted with a professional software package and the up-regulated genes at G1 late and G2 phase were analyzed according to gene database. Furthermore, the mRNA level of cyclin E, cyclin B, plk1 and STK15 in above mentioned nine points were measured by quatitative PCR.

<b>RESULTSb>2001 genes were detected to be available at all 9 points via software processing, out of which 959 appeared up-regulated or down-regulated. 379 genes showed to be up-regulated at late G1 (147) or G2 phases (232), 40 at S and M phases (also up-regulated at G1 late and G2 phases). The 147 up-regulated genes at G1 late phase are involved in DNA metabolism, transcription and translation, protein transportation, ubiquitination and signal transduction, etc. The 232 up-regulated genes in G2 phase are involved in RNA synthesis and processing, intracellular protein transportation, cytoskeleton synthesis, signal transduction, apoptosis and anti-apoptosis, transcription regulation, ubiquitination, mitosis regulation and oncogene expression, etc. The mRNA level of 4 genes detected by quantitative PCR during cell cycle was in agreement with that detected by microarray.

<b>CONCLUSIONb>During MKN45 cell cycling, the preparation for DNA synthesis and chromosome separation are conducted in G1 and G2, which are implicated in multiple genes, may be the main impetus of driving MKN45 cell cycle. Some of these genes may be related to tumor over-proliferation. The cDNA microarray technique has characteristic features such as reliability and can provide a great deal for future research on cell cycle related genes in gastric cancer.

Aurora Kinase A ; Aurora Kinases ; Cell Cycle ; genetics ; Cell Cycle Proteins ; genetics ; Cell Line, Tumor ; Cyclin B ; genetics ; Cyclin E ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; methods ; Protein-Serine-Threonine Kinases ; genetics ; Proto-Oncogene Proteins ; genetics ; RNA, Messenger ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; pathology