Humans ; Carcinoma, Squamous Cell/pathology* ; Chromosomal Proteins, Non-Histone/genetics* ; Ear, Middle/pathology* ; Poly-ADP-Ribose Binding Proteins ; Nuclear Proteins

OBJECTIVETo investigate the association between ERCC6 gene polymorphisms and peripheral blood lymphocyte DNA damage among the workers in coking plant.

METHODSBy cluster sampling, 379 coke oven workers having worked for 8 hours were included in the exposure group, 398 coke oven workers having rested for more than 16 hours were included in the recovery group, and 398 workers having never been exposed to polycyclic aromatic hydrocarbons (PAHs) in the same plant were included in the control group. Lymphocytes were separated from their peripheral venous blood, and single cell gel electrophoresis was used to evaluate DNA damage; TaqMan-MGB probes were used to analyze ERCC6 gene polymorphisms. PHASE 2.0.2 genetic analysis software was used to calculate the haplotypes.

RESULTSThe Olive tail moment (OTM) of lymphocytes in the exposure group was significantly higher than those in the recovery group and control group (-0.86±0.70 vs -1.14±0.68 and -1.13±0.65, P < 0.05). In the exposure group, for workers ≥37 years old, the OTM of lymphocytes in workers carrying CG+GG genotype at rs3793784 locus of ERCC6 gene was significantly lower than that in workers carrying CC genotype (P < 0.05); the OTM of lymphocytes in workers <37years old carrying CC genotype at rs3793784 locus of ERCC6 gene was significantly lower than that in workers ≥37 years old carrying CC genotype (P < 0.05); the OTMof lymphocytes in workers <37 years old carrying CG+GG genotype at rs3793784 locus of ERCC6 gene was significantly higher than that in workers ≥37 years old carrying CG+GG genotype (P < 0.05). For patients with internal exposure, in the 1-hydroxypyrene >4.36 ümol/L group, the OTM of lymphocytes in workers carrying AG+GG genotype was significantly higher than that in workers carrying AA genotype (P < 0.05).

CONCLUSIONDifferent genotypes of ERCC6 gene rs3793784 in peripheral blood lymphocytes of coke oven workers exposed to PAHs have different functions at different ages, suggesting that genotype may interact with age in population exposed to PAHs.

Adult ; Coke ; DNA Damage ; DNA Helicases ; genetics ; DNA Repair Enzymes ; genetics ; Genotype ; Humans ; Lymphocytes ; Middle Aged ; Occupational Exposure ; adverse effects ; Poly-ADP-Ribose Binding Proteins ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the effect of small interfering RNA（siRNA）for MSI-2 on the growth, apoptosis and NUMB expression of THP-1 cells.

METHODSThree siRNA for MSI-2 gene was designed and transfected into THP- 1 cells. The cell inhibition, colony formation and apoptosis were determined. The protein expression of NUMB, caspase- 3 and PARP were detected by Western blotting.

RESULTSAfter MSI- 2 expression of THP- 1 cells was down- regulated for 24 hours, cell inhibition of siRNA MSI-2 group was（47.89±7.64）%, obviously higher than that of negative control group（P=0.005）. After 9 days, cell colony count of siRNA MSI-2 group was 7.50±1.53, also lower than that of negative control group（35.75±7.46, P<0.001）. In addition, apoptotic rates of siRNA MSI- 2 group at 24 hours ［（15.22±1.52）%］and 48 hours［（33.83±3.96）%］were significantly higher than those of negative control group（P=0.008 and P=0.001, respectively）. Accordingly, activations of caspase-3 and PARP and increased NUMB were observed in siRNA MSI- 2 group.

CONCLUSIONsiRNA for MSI- 2 gene could increase the expressions of NUMB to inhibit the proliferation and induce apoptosis of THP-1 cells.

Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Humans ; Membrane Proteins ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism ; RNA Interference ; RNA, Small Interfering ; RNA-Binding Proteins ; genetics ; metabolism ; Transfection

This study was purposed to investigate the significance of mitosis checkpoint gene chfr expression in acute leukemia (AL). 2 ml of bone marrow were extracted from each of 46 AL patients and 10 normal donors as control and their mononuclear cells were isolated. Then, their chfr expression was detected by using RT-PCR and immunohistochemistry. Normal control blood samples were also analyzed. The results showed that in 15 out of 28 cases of acute non-lymphocytic leukemia and 13 out of 18 cases of acute lymphocytic leukemia expression of chfr gene mRNA and protein significantly decreased as compared with control. The cytogenetic analysis of patients with a decreased Chfr expression revealed abnormal chromosome. In conclusion, Chfr gene is a leukemia-related gene and may play an important role in leukemia pathogenesis.
Bone Marrow Cells ; metabolism ; Cell Cycle Proteins ; biosynthesis ; genetics ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Mitosis ; Neoplasm Proteins ; biosynthesis ; genetics ; Poly-ADP-Ribose Binding Proteins ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Ubiquitin-Protein Ligases

OBJECTIVE: To discover the relationship of transcriptional levels and promoter methylation status of CHFR gene in human nasopharyngeal carcinoma,to discuss the significance and epigenetic mechanism of CHFR inactivation in NPC, and to evaluate the feasibility of detecting methylated CHFR in nasopharyngeal swab as a means for diagnosis of NPC. METHOD: Transcriptional levels of CHFR was evaluated by RT-PCR. Methylation specific PCR was used to detect the methylation status of CHFR in NPC cells, normal nasopharyngeal epithelia, primary tumors and their paired nasopharyngeal swabs. Detailed methylation status was confirmed by bisulfite sequencing. NPC cells were treated by the methyltransferase inhibitor 5-aza-dC and the reactivation of CHFR was evaluated by RT-PCR. RESULT: CHFR transcription was inactivated in NPC. The methylation frequency in NPC primary tumors and their paired swabs were 65.5% and 63.8%, respectively, with a 86.2% concordance. Bisulfite sequencing revealed a dense methylation in NPC cells and primary tumors, but all the normal nasopharyngeal epithelia were unmethylated. CHFR expression were restored after 5-aza-dC treatment. CONCLUSION CHFR is epigenetically inactivated by promoter methylation in NPC. Detecting methylated CHFR can be served as a useful non-invasive means for diagnosis of NPC.
Aged ; Carcinoma ; Cell Cycle Proteins ; genetics ; DNA Methylation ; Epigenesis, Genetic ; Female ; Gene Silencing ; Humans ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Neoplasm Proteins ; genetics ; Neoplasm Staging ; Poly-ADP-Ribose Binding Proteins ; Promoter Regions, Genetic ; Ubiquitin-Protein Ligases

OBJECTIVETo identify potential mutations among three sisters from a Chinese family suspected with Cockayne syndrome for growth and psychomotor retardation, and to offer genetic counseling and prenatal diagnosis for the family.

METHODSG-banded karyotyping, microarray comparative genomic hybridization (CM-CGH), whole genome exon high-throughput sequencing and Sanger sequencing were employed to identify potential genetic variations for the three patients and their parents.

RESULTSWhole exome sequencing has identified two novel missense mutations, i.e., c.1595A>G (p.Asp532Gly) and c.1607T>G (p.Leu536Trp), in exon 7 of excision repair cross-complementing rodent repair deficiency, complementation group 6 (ERCC6) gene. Sanger sequencing confirmed that all of the three sisters have inherited one of the mutations (c.1607T>G) from their father and another (c.1595A>G) from their mother.

CONCLUSIONThree sisters have all been identified as double heterozygote for mutations c.1607T>G and c.1595A>G and were diagnosed with Cockayne syndrome.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child, Preschool ; Cockayne Syndrome ; diagnosis ; genetics ; DNA Helicases ; genetics ; DNA Repair Enzymes ; genetics ; Exons ; Female ; Heterozygote ; Humans ; Infant ; Male ; Molecular Sequence Data ; Pedigree ; Point Mutation ; Poly-ADP-Ribose Binding Proteins

OBJECTIVECockayne syndrome is a rare disease and difficult to be recognized. This study aimed to expand the knowledge of the clinical and molecular characteristics of the children with Cockayne syndrome (CS).

METHODClinical data of two siblings with classic CS of Guangzhou Women and Children's Medical Center from July 2013 to November 2014 were obtained and analyzed. The whole DNA of peripheral blood was collected from two CS siblings and their parents. Amplification of all exons and adjacent introns for ERCC6 gene was conducted using PCR, and measurement of reaction product was performed to find mutation sites by two-way sequencing.

RESULTTwo affected siblings were males, and came from unconsanguineous parents, 7 years and 5 months old and 4 years and 8 months old, respectively. They were in treatment because of developmental and mental retardation for years. When they were younger than one year of age, their heights and weight were within normal limits. However, poor growth of height and weight and psychomotor retardation appeared after one and a half years of age, as well as skin and eye sensitivity to sunshine, hearing impairment, optic nerve atrophy, microcephaly, and deep-set eyes. The proband's height was 90.8 cm, and weight 9.1 kg, head circumference 41 cm, and chest circumference 44 cm when he was taken to hospital. The elder brother of the proband had a height of 92 cm, weight 11.2 kg, head circumference 41 cm, and chest circumference 44 cm when he was taken to hospital. When the proband was four and a half years old, ventricular enlargement, hypomyelination, and brain atrophy were detected for his elder brother at 7 years of age by cranial MRI. MRS imaging indicated that damages occurred at the left and right sides of dorsal thalamus, lobus insularis, along with the left half circle of central neurons. Symmetrical calcification on bilateral basal ganglia was found on the brain CT scan. Pathogenic compound heterozygous c. 1357C > T (p.Arg453Ter) and c. 1607T > G (p.Leu536Trp) mutations of ERCC6 gene were identified in the two siblings which were separately inherited from their unaffected parents.

CONCLUSIONCS children are usually normal at birth, however, they have severe clinical characteristics such as poor growth, psychomotor retardation, cerebral injury, microcephalus, deep-set eyes, and skin sensitivity to sunshine. ERCC6 gene mutation usually occurs, and it is easy to misdiagnose CS as cerebral palsy, primary microcephaly, and so on.

Asian Continental Ancestry Group ; Child ; Child, Preschool ; Cockayne Syndrome ; genetics ; DNA Helicases ; genetics ; DNA Mutational Analysis ; DNA Repair Enzymes ; genetics ; Exons ; Heterozygote ; Humans ; Magnetic Resonance Imaging ; Male ; Mutation ; Poly-ADP-Ribose Binding Proteins ; Polymerase Chain Reaction ; Siblings

Chfr, a mitotic stress checkpoint gene, regulates a prophase delay in cells exposed to agents that disrupt microtubules, such as nocodazole and taxol. Chfr expression was ubiquitious in normal human tissues. It is very high conserved between human and mice. Preliminary sutdies indicated that Chfr expression was cell cycle regulated and it dependent on its ubiqitin ligase activity. The direct target of the Chfr pathway was Polo-like kinase 1 (Plk1). Ubiquitination of Plk1 by Chfr delayed the activation of the Cdc25C phosphatase and the inactivation of the Weel kinase, leading to a delay in Cdc 2 activation. The chfr gene was inactivated owing to lack of expression or by mutation in some human cancer cell lines examined. Normal primary cells and tumour cell lines that express wild-type chfr exhibited delayed entry into metaphase when centrosome separation was inhibited by mitotic stress. In contrast, the tumour cell lines that had lost chfr function entered metaphase without delay. Ecotopic expression of wild-type chfr restored the cell cycle delay and increased the ability of the cells to survive mitotic stress. Thus, chfr defines a checkpoint that delays entry into metaphase in response to mitotic stress. The progress of research on structure of Chfr gene and effects of Chfr protein was reviewed.
Cell Cycle ; genetics ; physiology ; Cell Cycle Proteins ; genetics ; metabolism ; physiology ; Humans ; Metaphase ; genetics ; physiology ; Mitosis ; genetics ; physiology ; Neoplasm Proteins ; genetics ; physiology ; Neoplasms ; genetics ; metabolism ; pathology ; Poly-ADP-Ribose Binding Proteins ; Prophase ; genetics ; physiology ; Protein-Serine-Threonine Kinases ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Ubiquitin-Protein Ligases

BACKGROUND AND OBJECTIVETranscriptional silencing induced by CpG island methylation is believed to be one of the important mechanisms of carcinogenesis. Checkpoint with fork head-associated and ring finger (CHFR) governs the transition from prophase to prometaphase in response to mitotic stress. This study was to analyze the relationship between the methylation of CHFR gene and the clinicopathologic features of gastric cancer, and the difference of results between methylation-specific polymerase chain reaction (MSP) and combined bisulfite restriction analysis (COBRA) in detecting aberrant methylation of CHFR gene in gastric cancer.

METHODSBoth MSP and COBRA methods were used to detect the promoter methylation of CHFR gene in gastric cancer specimens from 64 patients. The relationship between methylation status of CHFR gene and the clinicopathologic features of gastric cancer were analyzed using SPSS16.0.

RESULTSThe methylation rates of CHFR gene promoter were significantly higher in gastric cancer samples than in the corresponding paracancer normal gastric mucosa by MSP (51.6% vs. 18.8%, P < 0.001). However, there was no significant correlation between methylation status of CHFR gene and the clinicopathologic parameters of gastric cancer, including age, gender, tumor size, clinical stage, Borrman type, tumor invasion depth, differentiation, and lymph node metastasis (P > 0.05). Aberrant methylation of the CHFR gene was detected in 27 (42.2%) of the 64 specimens of gastric cancer using COBRA, which did not significantly differ from that using MSP (P > 0.05).

CONCLUSIONSAberrant methylation of the CHFR gene is a frequent event in the carcinogenesis of gastric cancer. Detecting the methylation of CHFR gene in gastric mucosa may conduce to the diagnosis of gastric cancer. No difference was found between MSP and COBRA in detecting promoter methylation of CHFR gene in gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Cell Cycle Proteins ; genetics ; DNA Methylation ; DNA, Neoplasm ; genetics ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Proteins ; genetics ; Neoplasm Staging ; Poly-ADP-Ribose Binding Proteins ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Sulfites ; chemistry ; Ubiquitin-Protein Ligases

OBJECTIVE: To explore the relationship between the level of expression and hypermethylation of the CHFR gene and the occurrence and development of laryngeal squamous cell carcinoma (LSCC). METHOD: The mRNA expression and promoter hypermethylation were detected by Realtime fluro-genetic quantitative PCR and methylation specific PCR in 50 LSCCs (LSCC group) and 15 normal laryngeal tissue (control group). RESULT: 1) CHFR mRNA was shown in the control group, while the mRNA was loss expression in the 2 LSCC (4%), and the level of mRNA expression was significantly lower in the LSCC group. The relative ratio was 0.50 +/- 0.12, which is 0.30 +/- 0.04 at the early stage of the LSCC and 0.70 +/- 0.21 at the advanced stage, respectively. The discrepancy had statistical significance (P<0.01). 2) The methylation rate of CHFR was 22% (11/50) in the LSCC tissues, which was not found in the normal tissues. The aberrant methylation of CHFR was observed in 10 of the patients at the stage I and stage II of LSCC , in 1 of the patients at the stage III, and was absent at the stage IV. There was significant difference between the aberrant methylation of CHFR and the stage of carcinoma (P<0.01). 3) The mRNA expression level of the aberrant methylation patients was 0.11 +/- 0.05, which was significantly lower than that of the unmethylation patients 0.75 +/- 0.13. Gene inactivation was observed in 2 of the 11 patients with the aberrant promoter methylation. The methylation was associated with the expression of mRNA, with the correlation coefficient 0.387 (P<0.05). CONCLUSION Hypermethylation of CHFR gene promoter is associated with loss or lower expression of CHFR mRNA in the LSCCs, and it may contribute to the occurrence and development of LSCC. The promoter aberrant methylation of CHFR may be one of the early diagnostic and therapeutic marker genes.
Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Cycle Proteins ; genetics ; CpG Islands ; DNA Methylation ; Female ; Gene Silencing ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; Neoplasm Staging ; Poly-ADP-Ribose Binding Proteins ; Promoter Regions, Genetic ; RNA, Messenger ; genetics ; Ubiquitin-Protein Ligases