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

OBJECTIVETo detect the correlation between the expression of topoisomerase 2 alpha (TOP2A) and the biological behaviors of human colorectal carcinoma.

METHODSImmunohistochemistry and real-time RT-PCR were used to detect the expression of TOP2A in colorectal carcinomas and normal mucosa.

RESULTSThe protein and mRNA expressions of TOP2A in the metastatic lymph nodes were significantly higher than those in matched primary lesions and normal tissues (P<0.05). No significant difference was found in TOP2A expressions between normal mucosa and colorectal carcinomas. The protein and mRNA expressions of TOP2A were significantly correlated to the lymph node metastasis and invasion depth (P<0.05), but not to the differentiation of the tumor (P>0.05).

CONCLUSIONTOP2A plays an important role in the invasion and metastasis of the colorectal carcinomas, and may serve as a valuable indicator for the diagnosis, treatment and the prognostic evaluation of the malignancy.

Antigens, Neoplasm ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; DNA Topoisomerases, Type II ; metabolism ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Male ; Poly-ADP-Ribose Binding Proteins

OBJECTIVE: To investigate the gene mutation occurved in AML patients with 29 kinds of fusion genes and 51 kinds of tumor gene. METHODS: Next-generation sequencing (NGS) was used to detected the 49 kinds of targeted gene. FLT3 internal tandem duplication (FLT3-ITD), CALR, NPM1 and CEBPA mutation were detected by DNA-based PCR and Sanger sequencing. Twenty-nine kinds of fusion genes were dected by multiplex nested RT-PCR. RESULTS: The total gene mutation rate was 91% (109/121) in all the 121 patients. On average, 2.1 mutated genes per patient were identified, among these 121 patients, coexistence of ≥ 3 mutations was frequent (34.7%). The most commonly mutated genes were NRAS (23.96%, n=29), followed by NPM1 (14.04%, n=17), CEBPA double mutations (14.04%, n=17), KRAS (11.57%, n=14)，FLT3-ITD (10.74%, n=13), CSF3R (10.74%, n=13), TET2 (9.92%, n=12) and IDH1 (9.1%, n=11). Overall, fusion genes were detected in 47 (37.3%) patients, including AML/ETO (n=12), CBFβ/MYH11 (n=11), PML/RARa (n=12), MLL rearranagement realated mutation MLL-X (n=10). TLS/ERG (n=1) and DEK/CAN (n=1) in an order of decreasing frequency. Patients with normal karyotype (NK)- AML exhibited more mutations in CEBPA, NPM1, TET2, RUNX1 and IDH1, comparing with abnormal karyotype patients. KRAS mutation in abnormal kayotype patients was significantly higher than that in normal kayotype patients (P=0.014). TP53 mutations were predominantly associated with complex cytogenetics (P=0.199). KRAS mutations were more frequent in core binding factor (CBF) acute myeloid leukemia (AML) and 11q23/MLL rearrangement leukemia, compared with NK-AML (P=0.006 and 0.003, respectively). KIT mutations predominated in CBF-AML (P=0.006). JAK2V617F mutations were detected in two patients and co-occurred with AML-ETO fusions. CONCLUSION At least one mutation is observed in more than 90% patients. On average, more than 2 mutated genes per patient are identified. Some gene mutations are associated with gene rearrangement.
Chromosomal Proteins, Non-Histone ; Genomics ; High-Throughput Nucleotide Sequencing ; Humans ; Leukemia, Myeloid, Acute ; Mutation ; Oncogene Proteins ; Poly-ADP-Ribose Binding Proteins ; Prognosis

OBJECTIVETo study the immunophenotype and differential diagnosis of Hodgkin's lymphoma (HL).

METHODFifty six cases originally diagnosed as HL were re-evaluated according to lymphoma classification of WHO 2000 on paraffin sections using SP immunohistochemistry.

RESULTSAmong the 56 cases, 47 met the WHO criteria for HL, 8 were NHL and 1 metastatic tumor. Of the 47 HL cases, 2 were nodular lymphocyte predominant HL (NLPHL), 43 classical Hodgkin's lymphoma (CHL) and 2 unclassified HL, and of the 8 cases reclassified as NHL, 6 were T-cell rich B-cell lymphoma (TCRBCL) and 2 anaplastic large cell lymphoma (ALCL). In NLPHL cases, L&H cells were CD(20)(+), CD(15)(-) and CD(30)(-); CD(57)(+) cells and small B-lymphocytes predominated the background infiltration. Diagnostic R-S cells and other tumor cells in 43 cases of CHL were positive for CD(30) (100%), CD(15) (81%) and CD(20) (7%). Six cases of TCRBCL were negative for CD(15) and CD(30). Two cases of ALCL were positive for CD(30), ALK-1 and CD(3), and negative for CD(15) and CD(20). The reactive inflammatory infiltration in CHL and TCRBCL was rich in TIA-1 positive cytotoxic lymphocytes, and CD(57)(+) cells were rarely encountered.

CONCLUSIONCombining the immunophenotype of tumor cells and background cells with morphologic criteria are more helpful for classification of HL, and discrimination between NLPHL, CHL and TCRBCL.

Antigens, CD ; analysis ; Diagnosis, Differential ; Hodgkin Disease ; immunology ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Immunophenotyping ; Membrane Proteins ; analysis ; Mucin-1 ; analysis ; Poly(A)-Binding Proteins ; Proteins ; RNA-Binding Proteins ; analysis ; T-Cell Intracellular Antigen-1

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

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 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

OBJECTIVE: To assess the value of m⁷G-lncRNAs in predicting the prognosis and microenvironment of colorectal cancer (CRC). METHODS: We screened m⁷G-lncRNAs from TCGA to construct an m⁷G-lncRNAs risk model using multivariate Cox analysis, which was validated using ROC and C-index curves. Calibration and nomogram were used to predict the prognosis of CRC patients. Point-bar charts and K-M survival curves were used to assess the correlation of risk scores with the patients' clinical staging and prognosis. CIBERSORT and ESTIMATE were used to explore the association between the tumor microenvironment and immune cell infiltration in patients in high and low risk groups and the correlation of risk scores with microsatellite instability, stem cell index and immune checkpoint expression. A protein-protein interaction network was constructed, and the key targets regulated by m⁷G-lncRNAs were identified and validated in paired samples of CRC and adjacent tissues by immunoblotting. RESULTS: We identified a total of 1722 m⁷G-lncRNAs from TCGA database, from which 12 lncRNAs were screened to construct the risk model. The AUCs of the risk model for predicting survival outcomes at 1, 3 and 5 years were 0.727, 0.747 and 0.794, respectively. The AUC of the nomogram for predicting prognosis was 0.794, and the predicted results were consistent with actual survival outcomes of the patients. The patients in the high-risk group showed more advanced tumor stages and a greater likelihood of high microsatellite instability than those in the low-risk group (<i>Pi> < 0.05). The tumor stemness index was negatively correlated with the risk score (<i>ri>=-0.19; <i>Pi>=7.3e-05). Patients in the high-risk group had higher stromal cell scores (<i>Pi>=0.0028) and higher total scores (<i>Pi>=0.007) with lowered expressions of activated mast cells (<i>ri>=-0.11; <i>Pi>=0.045) and resting CD4+ T cells (<i>ri>=-0.14; <i>Pi>=0.01) and increased expressions of most immune checkpoints (<i>Pi> < 0.05). ATXN2 (<i>Pi>= 0.006) and G3BP1 (<i>Pi>=0.007) were identified as the key targets regulated by m⁷G-lncRNAs, and their expressions were both higher in CRC than in adjacent tissues. CONCLUSION The risk model based on 12 m⁷G-lncRNAs has important prognostic value for CRC and can reflect the microenvironment and the efficacy of immunotherapy in the patients.
Biomarkers, Tumor/metabolism* ; Colonic Neoplasms ; DNA Helicases/metabolism* ; Gene Expression Regulation, Neoplastic ; Humans ; Microsatellite Instability ; Poly-ADP-Ribose Binding Proteins/metabolism* ; Prognosis ; RNA Helicases/metabolism* ; RNA Recognition Motif Proteins/metabolism* ; RNA, Long Noncoding/metabolism* ; Tumor Microenvironment

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