Cell Cycle ; Cell Cycle Proteins ; analysis ; antagonists & inhibitors ; physiology ; Cell Differentiation ; Cell Proliferation ; Humans ; Membrane Proteins ; analysis ; antagonists & inhibitors ; physiology ; Neoplasm Proteins ; analysis ; antagonists & inhibitors ; physiology ; Neoplasms ; enzymology ; therapy ; Protein Tyrosine Phosphatases ; analysis ; antagonists & inhibitors ; physiology

OBJECTIVETo study the influence of human cytomegalovirus (HCMV) infection on cell cycle and the expression of replication licensing factor Cdt1 in human embryonic lung fibroblastic (HEL) cells and to explore the pathogenesis of HCMV infection.

METHODSHEL cells were synchronized in the G0/G1 phase by the serum starvation method. The synchronized HEL cells were infected with HCMV, and those that were not subjected to HCMV infection were used as the control group. The HEL cells were harvested at 12, 24, 48, 72 and 96 hrs of HCMV infection. The cell cycle of HEL cells was detected by the flow cytometry. The expression of Cdt1 mRNA in HEL cells was determined by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe cells in the G1 phase in the control group was significantly more than in the HCMV-infected group 12 and 24 hrs after infection (P < 0.01). The expression of Cdt1 mRNA in the HCMV-infected group was significantly lower 12 and 24 hrs after infection but increased significantly 48 hrs after infection compared with the control group (P < 0.05). The expression of Cdt1 mRNA reached a peak at 12 hrs of infection in the control group, but at 48 hrs of infection in the HCMV-infected group, which markedly lagged behind the control group.

CONCLUSIONSHCMV infection arrests the cell cycle of HEL cells at the G1 phase. HCMV infection makes Cdt1 expression delay. HCMV infection can interfere cell cycle of HEL cells possibly through affecting the expression of Cdt1.

Cell Cycle ; Cell Cycle Proteins ; genetics ; Cells, Cultured ; Cytomegalovirus ; pathogenicity ; Embryo, Mammalian ; cytology ; Fibroblasts ; cytology ; metabolism ; Humans ; Lung ; cytology ; metabolism ; RNA, Messenger ; analysis

BACKGROUNDNucleostemin is essential for the proliferation and survival of stem and cancer cells, but it is unknown whether this newly identified molecule is involved in prostate cancer pathogenesis.

METHODSTotal RNA and protein were extracted from prostate cancer tissues and PC-3, LNCap and DU145 cell lines. The nucleostemin mRNA and protein expression were measured by RT-PCR and Western blot. Immunohistochemistry was also used to detect the nucleostemin protein expression in prostate cancer tissues and PC-3 cells. A nucleostemin specific, short hairpin RNA, expression plasmid was used to transfect PC-3 cells. The changes of nucleostemin gene were detected and the proliferative capacity of the cells was determined.

RESULTSNucleostemin was highly expressed in prostate cancer tissues and cell lines. Nucleostemin expression level in the silencer group PC-3 cells remarkably reduced. The proliferation rate of silencer group PC-3 cells decreased and the percentage of G1 stage cells increased. The neoplasm forming capacity in nude mice of the silencer group PC-3 cells decreased significantly.

CONCLUSIONSNucleostemin is highly expressed in prostate cancer tissues and cell lines. The proliferative capacity of PC-3 cells is remarkably reduced after silencing nucleostemin gene expression.

Animals ; Apoptosis ; Carrier Proteins ; analysis ; genetics ; physiology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; GTP-Binding Proteins ; Humans ; Male ; Mice ; Nuclear Proteins ; analysis ; genetics ; physiology ; Prostatic Neoplasms ; genetics ; pathology ; RNA, Messenger ; analysis

Cell Cycle ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; analysis ; DNA Damage ; Dactinomycin ; pharmacology ; Humans ; Viral Nonstructural Proteins ; physiology

OBJECTIVETo study the relationship between hRad17 mRNA expression and clinicopathologic factors and lymph node metastasis of gastric cancer, and to assess the significance of predicting the extent of lymph node metastasis and prognosis.

METHODShRad17 mRNA expression was examined in matched primary lesions, normal gastric mucosa and lymph node metastatic lesions among 52 gastric cancer patients by reverse transcription polymerase chain reaction (RT-PCR), polyacrylamide gel electrophoresis (PAGE) and silver stain with the relation between hRad17 mRNA expression and clinicopathologic factors analyzed. At the same time, hRad17 mRNA expressions in 5 gastric benign lesions and SGC7901 gastric carcinoma cell lines were also examined.

RESULTSThe primary tumor samples (88.4% positive) showed a significantly higher level of hRad17 expression compared with matched normal tissue (76.9% positive) (P = 0.014), so did the lymph node metastatic samples (94.2% positive) (P = 0.001). The hRad17 mRNA expression showed a low level in benign lesions, but very high in SGC7901 cell line. The hRad17 mRNA expression showed a higher level in patients with the number of lymph node metastasis above 15 than below 15 (P = 0.02), so did the diffused growth than the mass-like growth (P = 0.04).

CONCLUSIONThe method of PAGE and silver stain can improve the sensitivity of RT-PCR. The degree of lymph node metastasis and invasiveness of carcinoma cells are more serious in cases with hRad17 mRNA overexpression, and extensive lymph node dissection should be carried out for these patients. Examination of hRad17 expression by RT-PCR before surgery is indicated to arrive at an optimum treatment scheme and to estimate the prognosis.

Cell Cycle Proteins ; genetics ; Gastric Mucosa ; metabolism ; Humans ; Lymphatic Metastasis ; RNA, Messenger ; analysis ; Stomach Neoplasms ; metabolism ; pathology

CCCTC-binding factor (CTCF) is a highly conserved zinc finger protein and is best known as a transcription factor. It can function as a transcriptional activator, a repressor or an insulator protein, blocking the communication between enhancers and promoters. CTCF can also recruit other transcription factors while bound to chromatin domain boundaries. The three-dimensional organization of the eukaryotic genome dictates its function, and CTCF serves as one of the core architectural proteins that help establish this organization. The mapping of CTCF-binding sites in diverse species has revealed that the genome is covered with CTCF-binding sites. Here we briefly describe the diverse roles of CTCF that contribute to genome organization and gene expression.
Animals ; Cell Cycle Proteins/metabolism ; Chromosomal Proteins, Non-Histone/metabolism ; *Gene Expression Regulation ; Genome ; Humans ; Protein Binding ; Protein Interaction Maps ; Repressor Proteins/analysis/*metabolism

Adolescent ; Adult ; Aged ; Apoptosis ; Cell Cycle ; Cell Division ; Child ; Female ; Glioma ; chemistry ; pathology ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; analysis ; Middle Aged ; Neoplasm Proteins ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; analysis

OBJECTIVETo evaluate the roles of p27(kip1), p16 gene protein and proliferating cell nuclear antigen expression in nasopharyngeal carcinoma (NPC).

METHODSThe EnVision immunohistochemical method was used to detect the expression of p27(kip1), p16 gene protein and PCNA in 66 cases of non-keratinized carcinoma (NKC) and 25 cases of non-tumor nasopharyngeal tissue.

RESULTS(1) The positive expression rates of p27(kip1), p16 gene protein were 65%, 68% in NKC respectively. There were significant differences between NKC and non-tumor group (P < 0.05). (2) The expression of p27(kip1), p16 protein correlated with cranial nerve encroaching and the 5-year survival rates of the patients (P < 0.05), but had no significant correlation to lymph node metastases and clinical staging (P > 0.05). The expression of PCNA was related to clinical staging and to the patient's 5-year survival rates (P < 0.05), but not to lymph node metastases and cranial nerve encroaching (P > 0.05). (3) The positive expression of p27(kip1), p16 gene protein and PCNA were correlated.

CONCLUSIONThe results suggest that immunological labeling of p27(kip1), p16 gene protein and PCNA might be used to determine the prognosis of NKC.

Adult ; Aged ; Cell Cycle Proteins ; analysis ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; chemistry ; mortality ; pathology ; Prognosis ; Proliferating Cell Nuclear Antigen ; analysis ; Tumor Suppressor Proteins ; analysis

OBJECTIVETo analyze the clinical features and genotype in a 8-year-old boy with dyskeratosis congenita (DC).

METHODSWe reviewed the clinical data of the case and amplified 7 DC-related genes (including DKC1,TERT,TERC,TINF2,NOP10, NHP2 and WRAP53) using polymerase chain reaction for DNA sequence analysis to identify the abnormal exons.

RESULTSDNA sequence analysis showed a c.85-15T>C mutation in DKC1 gene of the patient. His mother was a carrier of the mutated gene and presented with partial clinical features such as abnormal nails.

CONCLUSIONThe mutation of c.85-15T>C in DKC1 gene was reported for the first time in China. The diagnosis of DC should be considered if a young patient presents with mucocutaneous abnormalities, bone marrow failure, cancer susceptibility and a family history of cancer. Early genetic tests can improve the diagnosis rates and reduce misdiagnosis and missed diagnosis.

Cell Cycle Proteins ; genetics ; Child ; China ; Dyskeratosis Congenita ; genetics ; pathology ; Exons ; Genotype ; Humans ; Male ; Mutation ; Nuclear Proteins ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVE: To investigate the profile and potential significance of PTEN and NBS1 mutations among patients with familial or at early onset breast cancer in Hunan province.
 METHODS: A total of 131 breast cancer patients with familial history or suffered from breast cancer at the age of less than 35 years old were included in this study. A comprehensive phosphatase and tensin homolog (PTEN) and nibrin (NBS1) mutation analysis was performed through denaturing high performance liquid chromatography (DHPLC) and subsequent DNA direct sequencing.
 RESULTS: Among 131 patients, a reported mutation IVS4+109insTCTTA in PTEN gene were identified in two patients. The mutation frequency of IVS4+109insTCTTA was 1.15%. Two mutations in PTEN gene, 225 A>C (Thr 160 Pro) and IVS5+13T>C, was firstly discovered. Another reported missense mutation was rs121909229 G>A (Arg 130 Gln). Three mutations were detected in NBS1 gene, of which IVS6+43A>G and IVS6+127A>G were firstly discovered and another reported synonymous mutations was rs1805794 G>C (Glu 185 Gln).
 CONCLUSION The novel mutations in PTEN and NBS1 might be specific to the familial and early-onset breast cancer of Chinese Hunan population.
Adult ; Asian Continental Ancestry Group ; Breast Neoplasms ; genetics ; Cell Cycle Proteins ; genetics ; China ; DNA Mutational Analysis ; Female ; Humans ; Mutation ; Nuclear Proteins ; genetics ; PTEN Phosphohydrolase ; genetics