Analysis of 7 cases of pediatric acute myeloid leukemia with DEK-NUP214 fusion gene / 中华儿科杂志

Objective: To investigate the clinical features, treatment regime, and outcome of pediatric acute myeloid leukemia (AML) with DEK-NUP214 fusion gene. Methods: The clinical data, genetic and molecular results, treatment process and survival status of 7 cases of DEK-NUP214 fusion gene positive AML children admitted to the Pediatric Blood Diseases Center of Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences from May 2015 to February 2022 were analyzed retrospectively. Results: DEK-NUP214 fusion gene positive AML accounted for 1.02% (7/683) of pediatric AML diagnosed in the same period, with 4 males and 3 females. The age of disease onset was 8.2 (7.5, 9.5) years. The blast percentage in bone marrow was 0.275 (0.225, 0.480), and 6 cases were M5 by FAB classification. Pathological hematopoiesis was observed in all cases except for one whose bone marrow morphology was unknown. Three cases carried FLT3-ITD mutations, 4 cases carried NRAS mutations, and 2 cases carried KRAS mutations. After diagnosis, 4 cases received IAE induction regimen (idarubicin, cytarabine and etoposide), 1 case received MAE induction regimen (mitoxantrone, cytarabine and etoposide), 1 case received DAH induction regimen (daunorubicin, cytarabine and homoharringtonine) and 1 case received DAE induction regimen (daunorubicin, cytarabine and etoposide). Complete remission was achieved in 3 cases after one course of induction. Four cases who did not achieved complete remission received CAG (aclarubicin, cytarabine and granulocyte colony-stimulating factor), IAH (idarubicin, cytarabine and homoharringtonine), CAG combined with cladribine, and HAG (homoharringtonine, cytarabine and granulocyte colony-stimulating factor) combined with cladribine reinduction therapy, respectively, all 4 cases reached complete remission. Six patients received hematopoietic stem cell transplantation (HSCT) after 1-2 sessions of intensive consolidation treatment, except that one case was lost to follow-up after complete remission. The time from diagnosis to HSCT was 143 (121, 174) days. Before HSCT, one case was positive for flow cytometry minimal residual disease and 3 cases were positive for DEK-NUP214 fusion gene. Three cases accepted haploid donors, 2 cases accepted unrelated cord blood donors, and 1 case accepted matched sibling donor. The follow-up time was 20.4 (12.9, 53.1) months, the overall survival and event free survival rates were all 100%. Conclusions: Pediatric AML with DEK-NUP214 fusion gene is a unique and rare subtype, often diagnosed in relatively older children. The disease is characterized with a low blast percentage in bone marrow, significant pathological hematopoiesis and a high mutation rate in FLT3-ITD and RAS genes. Low remission rate by chemotherapy only and very high recurrence rate indicate its high malignancy and poor prognosis. Early HSCT after the first complete remission can improve its prognosis.

m7G-lncRNAs are potential biomarkers for prognosis and tumor microenvironment in patients with colon cancer / 南方医科大学学报

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 (P < 0.05). The tumor stemness index was negatively correlated with the risk score (r=-0.19; P=7.3e-05). Patients in the high-risk group had higher stromal cell scores (P=0.0028) and higher total scores (P=0.007) with lowered expressions of activated mast cells (r=-0.11; P=0.045) and resting CD4+ T cells (r=-0.14; P=0.01) and increased expressions of most immune checkpoints (P < 0.05). ATXN2 (P= 0.006) and G3BP1 (P=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.

Analysis of Genomic Landscape in Patients with Acute Myeloid Leukemia / 中国实验血液学杂志

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.

Clinical and molecular analysis of two Chinese siblings with Cockayne syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>Cockayne 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).</p><p><b>METHOD</b>Clinical 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.</p><p><b>RESULT</b>Two 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.</p><p><b>CONCLUSION</b>CS 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.</p>

Genetic analysis for a family with Cockayne syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>G-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.</p><p><b>RESULTS</b>Whole 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.</p><p><b>CONCLUSION</b>Three sisters have all been identified as double heterozygote for mutations c.1607T>G and c.1595A>G and were diagnosed with Cockayne syndrome.</p>

Association of ERCC6 gene polymorphisms and DNA damage in lymphocytes among coke oven workers / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the association between ERCC6 gene polymorphisms and peripheral blood lymphocyte DNA damage among the workers in coking plant.</p><p><b>METHODS</b>By 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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSION</b>Different 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.</p>

Effects of 5-Aza-2'-deoxycitydine and trichostatin A on expression and methylation of CHFR in human laryngreal carcinoma cell line / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the effects of 5-Aza-2'-deoxycitydine(5-Aza-dC) and trichostatin A (TSA) on the expression and methylation of CHFR in human laryngeal carcinoma cell line.@*METHOD@#The mRNA expression and promoter hypermethylation and were detected by Realtime fluro-genetic quantitative PCR and methylation specific PCR in Hep-2 cell line, which were cultured in vitro and then treated with different concentrations of 5-Aza-dC and TSA.@*RESULT@#Compared with the control team, 5-Aza-dC alone reactivated expression of the CHFR in Hep-2 cell line (1.75 +/- 0.21). TSA had no effect on gene expression (1.05 +/- 0.13). The combined treatment with 5-Aza-dC and TSA increased gene expression (2.15 +/- 0.18). The cell lines showed a characteristic DNA methylation status. 5-Aza-dC and combined 5-Aza-dC and TSA resulted in demethylation of CHFR. In contrast, TSA alone did not affect the DNA methylation status of CHFR.@*CONCLUSION@#Hypermethylation of CHFR gene promoter is a common event in the occurrence and development of laryngeal carcinoma. The promoter aberrant methylation of CHFR is a main cause for down-expression of CHFR. After either treatment with 5-Aza-dC alone or in combination with TSA, the expression of CHFR is up-regulated duo to the reversal methylation. It can be a new idea to the therapy of laryngeal carcinoma.

Correlation of TOP2A gene expression and survival of breast cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to assess the TOP2A RNA expression and the relationship of TOP2A protein expression with metastasis-free interval in breast cancer patients.</p><p><b>METHODS</b>TOP2A expression was analyzed prior to surgery in 86 patients. The level of TOP2A gene amplification was analyzed by fluorescence in situ hybridization (FISH), its RNA expression level with RT-PCR, and their correlation with TOP2A protein expression was assessed by immunohistochemistry (IHC). The correlation between RNA expression level and metastasis-free interval in breast cancer patients was also analyzed.</p><p><b>RESULTS</b>Aberrations (amplification or deletion) of TOP2A copy number was observed in 25.6% (22/86) of the cases. TOP2A protein expression was detected in 66.3% (57/86) of the samples. There was a significant correlation between the TOP2A RNA expression and protein expression (P < 0.001). TOP2A gene expression was significantly associated with the metastasis-free interval in the breast cancer patients (P = 0.001). There was no significant correlation between TOP2A gene amplification and TOP2A protein expression (P = 0.211).</p><p><b>CONCLUSIONS</b>TOP2A RNA level is an objective and reliable prognostic indicator in breast cancer.</p>

Aberrant promoter hypermethylation of CHFR in nasopharyngeal carcinoma / 临床耳鼻咽喉头颈外科杂志

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.

G3BP: a promising target for cancer therapy / 药学学报

G3BP (Ras-GTPase-activating protein SH3 domain binding protein), a protein which binds to RasGAP SH3 domain, belongs to RNA-binding protein family, implicating in the downstream of Ras signaling. G3BP harbors the activities of endoribonuclease and DNA helicase, and can induce stress granules formation. G3BP plays a general role in the signal pathways of cell proliferation, differentiation, apoptosis and RNA metabolism. It has been shown to be over-expressed in a number of human malignancies and has a close relationship with tumor invasion and metastasis. Given that it has been implicated in several pathways that are known to be involved in cancer biology, G3BP may provide a new target for cancer therapy.

Promoter methylation of CHFR gene in gastric carcinoma tissues detected using two methods / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Transcriptional 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.</p><p><b>METHODS</b>Both 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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSIONS</b>Aberrant 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.</p>

Expression of topoisomerase II alpha in human colorectal carcinoma and its significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the correlation between the expression of topoisomerase 2 alpha (TOP2A) and the biological behaviors of human colorectal carcinoma.</p><p><b>METHODS</b>Immunohistochemistry and real-time RT-PCR were used to detect the expression of TOP2A in colorectal carcinomas and normal mucosa.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSION</b>TOP2A 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.</p>

Study of mRNA expression level and hypermethylation of CHFR promoter in the laryngeal squamous cell carcinoma tissue / 临床耳鼻咽喉头颈外科杂志

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.

Topoisomerase IIalpha and HER2/neu gene alterations and their correlation in pancreatic ductal adenocarcinomas / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the changes of topoisomerase IIalpha (TOP2A) and HER2/neu genes in pancreatic ductal adenocarcinomas of Chinese patients, and to determine their roles during carcinogenesis and tumor progression.</p><p><b>METHODS</b>Expressions of TOP2A and HER2/neu proteins were detected by using immunohistochemistry, while gene amplifications of TOP2A and HER2/neu were assessed by using multi-color fluorescence in situ hybridization (FISH). All the samples were of paraffin embedded and 10% formalin fixed tissue, including 26 cases of pancreatic ductal adenocarcinomas with adjacent non-neoplastic pancreatic tissues, 10 cases of chronic panreatitis, and 10 cases of normal pancreas. The correlation between TOP2A and HER2/neu gene status was analyzed.</p><p><b>RESULTS</b>By immunohistochemistry, the nuclear positive index of TOP2A in pancreatic ductal adenocarcinomas varied from 0.5% to 70%, and the positive rate of HER2/neu in pancreatic ductal adenocarcinomas was 46.2% (12/26). By FISH, 9/10 TOP2A amplified adenocarcinomas showed TOP2A and HER2/neu gene coamplification, while one case with HER2/neu gene amplification adenocarcinoma showed no TOP2A amplification. No expression of TOP2A, HER2/neu proteins and no amplification of TOP2A and HER2/neu gene were detected in adjacent non-neoplastic pancreatic tissues, chronic pancreatitis tissues and normal pancreas. No relationship was found between protein expression and gene amplification of TOP2A and HER2/neu (P > 0.05). TOP2A gene amplification was significantly correlated with HER2/neu gene amplification (P < 0.01).</p><p><b>CONCLUSIONS</b>Protein expression of TOP2A and HER2/neu are not associated with the gene amplification. There is a significant correlation between TOP2A amplification and HER2/neu gene amplification. Co-amplification of TOP2A and HER2/neu may play an important role in the carcinogenesis and progression of pancreatic carcinoma. Evaluation of the status of TOP2A and HER2/neu may be helpful to achieve target therapy of pancreatic carcinoma.</p>

Expression of Ras-GTPase-activating protein SH 3 domain binding protein and osteopontin in esophageal squamous carcinoma and their effects on prognosis / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the expressions and significances of Ras-GTPase-activating protein SH 3 domain binding protein(G3BP) and osteopontin (OPN) proteins in esophageal squamous carcinoma (ESC).</p><p><b>METHODS</b>The expressions of G3BP and OPN proteins in 80 cases of ESC were detected by immunohistochemistry. The relationships between the 2 protein expression and tumor size, differentiation degree, TNM stage, lymph node metastasis and prognosis of ESC were also explored.</p><p><b>RESULTS</b>(1) The positive expression rate of G3BP in ESC was 71.3%, and the rate in lymphoid metastatic group was significantly higher than that in non lymphoid metastatic group (Z=-2.283, P=0.022), but no relations were found between G3BP expression and diameter of tumor, differentiation and TNM grade (P>0.05). The G3BP positive expression group had shorter survival time than G3BP negative expression group (P=0.000). (2) The positive expression rate of OPN in ESC was 100%, and the degree of OPN expression was correlated with the differentiation (chi(2)=10.766, P=0.005) and lymphoid metastasis (Z=-2.289, P=0.022), but no relationship was found between the diameter of tumor and TNM grade (P>0.05). The expression of OPN were significantly related to survivals in a negative time-dependent manner in ESC patients (P=0.000). (3) The expression of G3BP protein correlated positively with the degree of OPN expression in ESC tissue (r(s)=0.376, P=0.001).</p><p><b>CONCLUSIONS</b>The expressions of G3BP and OPN proteins have a close relationship with lymphoid metastasis and survival in ESC patients. G3BP and OPN proteins can be considered as predictors of prognosis in ESC patients.</p>

Analysis of expression of mitosis checkpoint gene chfr in bone marrow cells of acute leukemia patients / 中国实验血液学杂志

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.

Monoclonal antibody against G3BP: preparation, characterization and its application in analysis of human tumors / 中华病理学杂志

<p><b>OBJECTIVE</b>To better understand the molecular mechanism of tumorigenesis and progression, the monoclonal antibody against G3BP (Ras-GAP SH3 binding protein), which serves as an important downstream effector of Ras signaling, was prepared, characterized and utilized in analysis of various human tumors.</p><p><b>METHODS</b>By using the prokaryotic expression vector pGEX-5X1, GST-G3BP fusion protein was expressed in E. coli BL21 under induction of IPTG. Purified GST-G3BP fusion protein was used to immunize BALB/c mice. The monoclonal antibody against G3BP was produced through conventional hybridoma method and characterized by ELISA, Western blot and immunohistochemical staining.</p><p><b>RESULTS</b>A hybridoma cell line secreting anti-G3BP IgG1 subtype antibody was obtained. Western blot and competitive inhibition assay showed that the antibody was G3BP-specific. Immunohistochemical staining demonstrated that G3BP was over-expressed in formalin-fixed and paraffin-embedded tissues of some human tumors, such as lung cancer, colon cancer, gastric cancer and breast cancer. In breast cancer specimens, the degree of G3BP expression correlated positively with the presence of lymph node metastasis and c-erbB2 expression.</p><p><b>CONCLUSIONS</b>The G3BP-specific monoclonal antibody derived from recombination protein can be used in ELISA, Western blot and immunohistochemical assay. It may provide an important tool in analysis of G3BP in in vitro and in vivo experiments. Besides, G3BP may serve as another prognostic marker for breast cancer.</p>

Advance of study on effects of Chfr gene of mitosis prophase checkpoint--review / 中国实验血液学杂志

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.