In recent years, with the improvement of the sensitivity of examination equipment and the change of people's living environment and diet, the rate of thyroid cancer has risen rapidly, which has increased nearly five folds in 10 years. The pathogenesis, clinical manifestation, biological behavior, treatment and prognosis of thyroid carcinoma of different pathological types are obviously different. Papillary thyroid carcinoma (PTC) can develop at any age, which accounts for about 90% of thyroid cancer. It progresses slowly and has favourable prognosis, but lymph node metastasis appears easily. Whether PTC is accompanied by lymph node metastasis has an important impact on its prognosis and outcome. The Raf murine sarcoma viral oncogene homolog B(BRAF)gene mutation plays a crucial role in PTC lymph node metastasis. Having an in-depth understanding of the specific role and mechanism of BRAF gene mutation in PTC is expected to provide new ideas for diagnosis and treatment of PTC.
Animals ; Carcinoma, Papillary/genetics* ; Humans ; Lymphatic Metastasis ; Mice ; Mutation ; Oncogenes ; Proto-Oncogene Proteins B-raf/genetics* ; Thyroid Cancer, Papillary/genetics* ; Thyroid Neoplasms/genetics*

OBJECTIVETodelineate the clinical and genetic features of a patient with myeloproliferative neoplasm (MPN) in association with PDGFRA and EVI1 genes rearrangements.

METHODSClinical data of the patient was collected. Conventional cytogenetics, fluorescence in situ hybridization (FISH) and nested PCR were carried out for the patient.

RESULTSThe patient has featured recurrent rash, joint pain, and intermittent fever. Laboratory tests showed hyperleukocytosis and marked eosinophilia. Physical examination revealed splenomegaly. His karyotype was 46,XY,t(3;5)(q26;q15)[6]/46,XY[10]. FISH assay showed that both PDGFRA and EVI1 genes were rearranged. Molecular studies of the mRNA suggested that there was a in-frame fusion between exon 12 of the PDGFRA gene and exon 9 of the FIP1L1 gene. Imatinib was initiated at a dosage of 200 mg, and after 10 months, the signal of the FIP1L1-PDGFRA fusion gene was undetectable in bone marrow sample. However, the expression of EVI1 mRNA was stable, with no significant difference found between the patient and 10 healthy controls.

CONCLUSIONMPN in association with PDGFRA and EVI1 genes rearrangements have unique clinical and genetic features. Genetic testing is helpful for early diagnosis. Imatinib may be effective for the treatment.

Antineoplastic Agents ; therapeutic use ; Base Sequence ; Chromosome Banding ; Chromosomes, Human, Pair 3 ; genetics ; Chromosomes, Human, Pair 5 ; genetics ; DNA-Binding Proteins ; genetics ; Gene Rearrangement ; Humans ; Imatinib Mesylate ; therapeutic use ; In Situ Hybridization, Fluorescence ; Karyotyping ; MDS1 and EVI1 Complex Locus Protein ; Male ; Myeloproliferative Disorders ; drug therapy ; genetics ; Proto-Oncogenes ; genetics ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; Transcription Factors ; genetics ; Translocation, Genetic ; Treatment Outcome ; Young Adult

OBJECTIVETo analyze the clinical features and prognosis of acute leukemia patients with the mixed lineage leukemia（MLL）gene rearrangements AF10 positive.

METHODS6 cases with MLL-AF10 positive were analyzed retrospectively, related literatures were reviewed to clarify MLL-AF10 patients'clinical features and prognosis.

RESULTSThe median age of 6 cases was 19.5 years old, 5 patients with fever onset, the onset white blood cells of 4 patients were less than 10×10⁹/L. 5 cases were as M₅ and 1 case M₄ according to FAB classification, the level of fusion gene（RQ-PCR）was 0.23%-22.60% when diagnosed, 4 cases had concomitant WT1 gene mutation, flow cytometry disclosed myeloid phenotype. Of 5 evaluated patients achieved the first complete remission after conventional chemotherapy, 2 cases of complex karyotype died, one case died of sepsis in induction, another died from failing of transplantation. 4 out of 5 transplant recipients gained long term survival.

CONCLUSIONThe MLL-AF10 positive patients were mostly young men, the majority FAB classification was M5 or M4, often onset with fever, low white blood cells and low level of fusion gene, usually associated with WT1 mutation. Conventional chemotherapy produced a high response rate, but easy to relapse, while the complex karyotype had a poor prognosis, allo-HSCT may have the potential to improve the prognosis of MLL-AF10 positive patients.

Acute Disease ; Chromosomes, Human, Pair 11 ; Female ; Flow Cytometry ; Gene Rearrangement ; Humans ; Leukemia ; diagnosis ; genetics ; Male ; Myeloid-Lymphoid Leukemia Protein ; genetics ; Phenotype ; Polymerase Chain Reaction ; Prognosis ; Proto-Oncogenes ; Recurrence ; Remission Induction ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the effects of trichloroethylene (TCE) toxicity on the normal human liver cells (L02 cells) and hepatocytes with CYP2E1 gene overexpression which was constructed through molecular cloning technology in our laboratory, then to explore the roles of CYP2E1 gene in TCE toxicity.

METHODSL02 cells and hepatocytes with CYP2E1 overexpression were treated with various doses of TCE (0,0.25, 0.5, 1.0, 2.0, 4.0 mmol/L) for 12h, the expression of apoptosis genes (Bcl-2, Caspase-3, Caspase-8, Caspase-9) and oncogenes (c-fos, c-myc, k-ras, p53) were determined by real-time fluorescent PCR.

RESULTSBcl-2 mRNA expression levels increased significantly in normal liver cells and CYP2E1-overexpressing cells after TCE treatment, Bcl-2 levels were 20%∼50%higher in CYP2E1-overexpressing cells than in L02 liver cells at doses of 0.25∼2.0 mmol/L TCE. Caspase-3, Caspase-8 and caspase-9 mRNA expression increased by 30%∼600% in CYP2E1-overexpressing cells at doses of 0.5∼4.0 mmol/L TCE when compared with L02 cells (P < 0.01). Additionally, c-fos, k-ras and c-myc mRNA expression levels were 25%∼120% higher in CYP2E1-overexpressing cells than in L02 cells (P < 0.01), p53 mRNA expression levels were lower 10%∼50% in CYP2E1-overexpressing cells than in L02 cells (P < 0.05 or P < 0.01).

CONCLUSIONSThere were significant differences for apoptosis gene and oncogene expression levels between normal liver cells and CYP2E1-overexpressing cells after they were treated with TCE, these findings indicated that CYP2E1 might play an important role in TCE metabolism in vivo.

Apoptosis Regulatory Proteins ; Caspase 3 ; Caspase 8 ; Caspase 9 ; Cytochrome P-450 CYP2E1 ; genetics ; Gene Expression ; Hepatocytes ; drug effects ; Humans ; Liver ; Proto-Oncogenes ; genetics ; RNA, Messenger ; Trichloroethylene ; toxicity

OBJECTIVETo detect the frequency of ROS1 gene rearrangement in non-small cell lung cancer ( NSCLC) patients by FISH, and to analyze the relationship between ROS1 gene rearrangement and clinical features (including age, sex, stage, histology, smoking history) with NSCLC.

METHODSThe ROS1 gene rearrangement in histological sections of 1 652 NSCLC tissues was detected by FISH. The extracted RNA was amplified and the sequences were analyzed by Sanger sequencing for ROS1-positive samples.

RESULTSROS1 rearrangement was identified in 53 specimens (3.2%) from the 1 652 NSCLC tissues. Among these positive cases, 15 were CD74-ROS1, 13 were SLC34A2-ROS1, 13 were SDC4-ROS1 and 12 were TPM3-ROS1. The frequency of ROS1 rearrangement was significantly higher in never-smoking patients (49 cases) than in smokers (4 cases) (P < 0.05). Patients with ROS1-positive NSCLC tended to be younger and there was no significant difference in sex (P > 0.05). All of the ROS1-positive samples were adenocarcinomas, with a tendency toward higher clinical stage (P < 0.05).

CONCLUSIONSROS1 rearrangement has diversity, and may be defined as a new molecular subtype of NSCLC. ROS1 rearrangement tends to occur in younger, and never-smoker lung adenocarcinoma patients.

Adenocarcinoma ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; Gene Rearrangement ; Humans ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; Oncogenes ; Protein-Tyrosine Kinases ; genetics ; Proto-Oncogene Proteins ; genetics ; metabolism

OBJECTIVETo study the clinical characteristics of ecotopic viral integration site-1 (EVI1) and BCR/ABL positive childhood leukemia.

METHODSClinical data of four children with EVI1 and BCR/ABL positive leukemia and eight children with BCR/ABL positive but EVI1 negative chronic myeloid leukemia (CML) were retrospectively analyzed.

RESULTSIn the four children with EVI1 and BCR/ABL positive leukemia, two were initially diagnosed with chronic phase of CML, one with accelerated phase of CML and one with high-risk acute lymphoblastic leukemia (ALL). There were no significant differences in clinical characteristics at diagnosis between the patients with EVI1 and BCR/ABL positive leukemia and BCR/ABL positive but EVI1 negative leukemia. CD33 and CD38 were highly expressed and t(9;22) abnormality was present in all patients with EVI1 and BCR/ABL positive leukemia. Two of the 3 children with EVI1 and BCR/ABL positive CML achieved complete remission one or three months after treatment. Acquired negative status conversion occurred for EVI1 but not BCR/ABL in one CML case. The 3 children with EVI1 and BCR/ABL positive CML survived 20, 13 and 14 months, respectively, without recurrence. The child with EVI1 and BCR/ABL positive ALL failed to achieve complete remission after the first course of treatment and discontinued further treatment.

CONCLUSIONSCo-expression of EVI1 and BCR/ABL fusion gene can be found in childhood CML and ALL. The relatively rare leukemia has not significant difference respect to clinical characteristics. Prognosis of the disease needs to be determined by clinical studies with a larger sample size.

Child ; DNA-Binding Proteins ; genetics ; Female ; Genes, abl ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; MDS1 and EVI1 Complex Locus Protein ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Prognosis ; Proto-Oncogenes ; genetics ; Retrospective Studies ; Transcription Factors ; genetics

OBJECTIVETo study the expression of ecotropic viral integration site (EVI1) gene in childhood acute myeloid leukemia (AML) and the clinical features of EVI1-positive children with AML.

METHODSThe clinical data of EVI1-positive children with AML were collected and analyzed. RT-PCR and real-time quantitative PCR were used for qualitative and quantitative analysis of expression of EVI1. Flow cytometry (FCM) was used for determining the immunophenotypes of bone marrow cells. Multiparameter FCM was used for monitoring minimal residual disease. The karyotypes were determined.

RESULTSOf 241 children with AML, 33 (13.7%) were positive for EVI1 expression. There were no significant differences in age at first visit as well as the white blood cell count, hemoglobin level, and platelet count in peripheral blood between EVI1-positive and EVI1-negative children with AML (P>0.05), but EVI1-positive children had a significantly increased proportion of females compared with EVI1-negative children (P<0.05). The change in EVI1 expression was not synchronous with clinical remission and the change of MRD: some children had clinical remission or negative conversion of MRD before negative conversion of EVI1, while some had negative conversion of EVI1 before clinical remission or while MRD showed positive. EVI1 gene was usually co-expressed with other fusion genes. CD33 (100%), CD38 (88%), and HLADR (76%) were highly expressed in EVI1-positive children with AML. Abnormal chromosome structure or number was found in 15 patients. Compared with EVI1-negative children, EVI1-positive children had significantly lower complete remission rates after the first course of treatment (P<0.05).

CONCLUSIONSEVI1-positive children with AML have a poor short-term prognosis. In the development of AML, the activation of EVI1 gene is not isolated, but the result of interactions with other genes or chromosome abnormalities, and the mechanism of activation and its function need further study.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; DNA-Binding Proteins ; genetics ; Female ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Immunophenotyping ; Infant ; Leukemia, Myeloid, Acute ; genetics ; immunology ; MDS1 and EVI1 Complex Locus Protein ; Male ; Neoplasm, Residual ; Prognosis ; Proto-Oncogenes ; genetics ; Transcription Factors ; genetics

Myelodysplastic syndrome (MDS) is highly heterogeneous clonal hematological malignancy, having a high rate of progression to acute myeloid leukemia (AML). With the rapid development of molecular biological techniques, plenty of gene mutations were found to have close relationships with the transformation from MDS to AML. SRSF2 is a RNA splicing-related gene, which mutation may prompt a poor prognosis, and have a higher rate of progressing to AML. DNMT3A plays an important role in DNA methylation, its mutation often indicate a worse overall survival and a more rapid progression to AML. ASXL1 regulates the synthesis of histone, which frameshift mutations are molecular marks of an adverse outcome. IDH contains IDH1 and IDH2, which are related with the Krebs cycle. Patients with IDH1 mutation have a shorter overall survival and a higher risk of AML transformation than that of patients with wild-type IDH1, while IDH2 was a poor prognostic factor for overall survival in patients with lower-risk MDS. Another gene related with DNA methylation is TET2, which is the most frequently mutated gene in MDS known so far and it may act as tumor-suppressor gene, but the opinions on its impact on patients' outcomes are still controversial. Some studies show that its mutations relate to a shorter time to progression to AML. Because of the differentiations in patients' races, regions and clinical characteristics, the results of different studies are varied. In this review, the recent advances on these related genes are summarized.
DNA (Cytosine-5-)-Methyltransferases ; genetics ; DNA-Binding Proteins ; genetics ; Genotype ; Humans ; Isocitrate Dehydrogenase ; genetics ; Leukemia, Myeloid, Acute ; genetics ; pathology ; Myelodysplastic Syndromes ; genetics ; pathology ; Nuclear Proteins ; genetics ; Oncogenes ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; Ribonucleoproteins ; genetics ; Serine-Arginine Splicing Factors

In order to prove that ectopic over-expression of Pim-2 could induce malignant transformation of human liver cell line L02, three groups of cells were set up including human liver cell line L02 (L02), L02 cells transfected with Pim-2 gene (L02/Pim-2) and L02 cells transfected with empty-vector (L02/Vector). Pim-2 expression levels were detected. The morphology, proliferation level, apoptosis rate and migration ability of the cells were detected respectively. Then the cells were subcutaneously inoculated into athymic mice and the microstructures of the neoplasm were observed. Compared with the controls, Pim-2 expression levels were significantly higher in L02/Pim-2 cells (P<0.05), and their morphology had obvious malignant changes. They also showed a significantly increased proliferation rate (P<0.05) and migration capacity (P<0.05), as well as a significantly decreased apoptosis rate (P<0.05). Only the athymic mice inoculated with L02/Pim-2 cells could generate neoplasm, and the morphology of the neoplasm coincided with that of the hepatoma. The results manifest that ectopic Pim-2 gene could be stably expressed in L02/Pim-2 cells. Both the morphological and biological changes of L02/Pim-2 cells demonstrate the trend of malignant transformation. L02/Pim-2 cells could generate hepatoma in athymic mice. In conclusion, Pim-2 could induce malignant transformation of human liver cell line L02.
Animals ; Apoptosis ; Cell Line ; Cell Movement ; Cell Proliferation ; *Cell Transformation, Neoplastic ; Humans ; Liver/pathology/physiology ; *Liver Neoplasms/genetics/pathology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; *Oncogenes ; Protein-Serine-Threonine Kinases/genetics/*metabolism ; Proto-Oncogene Proteins/genetics/*metabolism

The aim of study was to investigate the inhibitory effect of small interfering RNA on evi1 gene expression and biological characteristics in HEL cells and its mechanism. 3 siRNA (siRNA-1, siRNA-2, siRNA-3) specific for evi1 gene were synthesized and transfected into HEL cells in vitro. Experiments were divided into test and control groups. MTT method was used to assay the inhibitory effect of siRNA on cell proliferation; semiquantitative RT-PCR was used to detect the expression of evi1 gene mRNA; the cell viability was determined by trypan blue dye test; the change of cell cycle and apoptosis of cells were analyzed by flow cytometry. The results showed that siRNA-1 had strongest effect, and inhibitory effect was most obvious at 48 hours after transfection. When the concentration of siRNA raised to 120 nmol/L, the inhibitory rate reached to the peak. The inhibitory rate of siRNA-1 on proliferation of HEL cells, relative expression level of evi1 mRNA and cell viability at 48 hours after transfection were 72.22 ± 2.80%, 27.31 ± 1.11% and 26.05 ± 2.49%, which had significant difference from other groups (p < 0.001). The siRNA resulted in arrest of cell cycle at G(0)/G(1) phase, the cell amount at S phase obviously decreased, the apoptotic rate of HEL cells obviously increased (p < 0.01). It is concluded that the siRNA specific for evi1 gene can suppress the proliferation of HEL cells, reduce the expression of evi1 mRNA, decrease the cell viability, arrest the cell cycle at G(0)/G(1) phase, suppress cell mitosis, and promote cell apoptosis.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; MDS1 and EVI1 Complex Locus Protein ; Proto-Oncogenes ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transcription Factors ; metabolism