Effect of multi-target protein tyrosine kinase inhibitor Ponatinib on proliferation,adhesion and migration ability of human liver cancer cell line SK-Hep-1 / 中国生物制品学杂志

@#Objective To investigate the effect of a multi-target protein tyrosine kinase inhibitor,Ponatinib,on proliferation,homogeneity adhesion and migration ability of human liver cancer cell line SK-Hep-1.Methods SK-Hep-1 cells were cultured routinely and added with 24 tyrosine kinase inhibitors such as Ponatinib respectively,and the effect of Ponatinib on the survival and proliferation of SK-Hep-1 cells was detected by MTT assay.SK-Hep-1 cells were cultured routinely until the fusion degree reached 90%,then added with 0.1,0.5 and 1.0 μmol/L Ponatinib respectively,and the control group(without Ponatinib) was set up.The effect of Ponatinib on adhesion ability of SK-Hep-1 cells was detected by cell slow aggregation assay and dissociation assay,while the effect on migration ability by scratch test,and the effect on E-cadherin protein expression in SK-Hep-1 cells by Western blot.Results All 24 tyrosine kinase inhibitors inhibited SK-Hep-1 cells,among which Ponatinib showed the strongest inhibitory effect with a IC_(50) of(0.288±0.044) μmol/L.Compared with the control group,the number of cell mass(t=16.143,44.002 and 44.853 respectively,each P <0.001) and N_(TC)/N_(TE) [ratio of single cell number(N) after digestion by trypsin containing EDTA(TE) and CaCl_2(TC)](t=4.276,10.625 and 27.571 respectively,each P <0.05) decreased significantly and E-cadherin protein expression increased significantly(t=-3.757,-4.561and-6.922 respectively,each P <0.05) in 0.1,0.5 and 1.0 μmol/L Ponatinib groups;Scratch migration rate significantly decreased in 0.5 and 1.0 μmol/L Ponatinib groups(t=6.272～16.733 respectively,each P <0.01),while there was no significant difference in 0.1 μmol/L Ponatinib group(t=0.473 and 0.872 respectively,each P> 0.05) after 24 h and 48 h of scratch.Conclusion Ponatinib inhibited proliferation and migration of SK-Hep-1 cells and promoted cell adhesion.

Expression of protein tyrosine kinase 7 and receptor tyrosine kinase-like orphan receptor 2 in the brainstem of amyotrophic lateral sclerosis transgenic mice with hSODl-G93A mutation / 解剖学报

Objective To investigate the relationship between changes in protein tyrosine kinase 7 (PTK7) and receptor tyrosine kinase-like orphan receptor 2 ( ROR2) expression in the brainstem and the pathogenesis of amyotrophic lateral sclerosis (ALS). Methods Forty-four human superoxide dismutase 1( hSODl)-G93A mutant ALS transgenic mice were selected, and an equal number of wild-type littermates was used as control. The brainstems were isolated at da)' 70, day 95, day 108 and da)' 122 after birth, and the morphology of frypoglossal nucleus (12N) and nucleus of facial nerve(7N) neurons in the brainstem of the model mice were observed by Nissl staining. The mRNA and protein expression of PTK7 and ROR2 were detected by RT-PCR and Western blotting respectively, and the cellular localization and distribution of PTK7 and ROR2 in 12N and 7N were observed by immunofluorescence double-labeling technique. Results The result of Nissl staining showed that Nissl bodies in the neurons reduced distinctly with vacuolar degeneration of neurons, cell body atrophy and nuclear volume reduction in the 12N and 7N brainstems of ALS transgenic mice. RT-PCR result indicated that ROR2 and PTK7 mRNA level in the brainstem of ALS transgenic mice were up-regulated at da)' 70, da)' 95, day 108 and day 122 compared with wild-type littermates. Western blotting result showed that PTK7 protein was up-regulated at day 70, day 95, day 108 and day 122, ROR2 protein was up-regulated at day 70, day 95, day 108, and down-regulated at day 122 in the brainstem of ALS transgenic mice compared with wild-type littermates. Immunofluorescence result showed that ROR2/neuronal nuclei (NeuN)and PTK7/NeuN double positive cells, ROR2/glial fibrillary acidic protein (GFAP) and PTK7/GFAP double positive cells were observed in the 12N and 7N of the brainstem of ALS transgenic mice and wild-type mice, suggesting that ROR2 and PTK7 were expressed both in neurons and astrocytes. Conclusion PTK7 and ROR2 are abnormally expressed in the brainstem of ALS transgenic mice, which is closely related to the pathogenesis of ALS.

Expression and clinical significance of Janus protein tyrosine kinase/signal transducer and activator of transcription signaling pathway in primary gouty arthritis / 中华风湿病学杂志

Objective:To explore the expression and clinical significance of Janus protein tyrosine kinase/signal transducer and activator of transcription (JAK3/STAT5) signaling pathway in peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:Peripheral blood, clinical data and laboratory tests were collected from 50 patients with acute gout (AG), 50 patients with intermittent gout (IG) and 50 healthy controls (HC). Quantitative real-time-polymerase chain reaction (RT-qPCR) was used to detect mRNA expression level of JAK3/STAT5 related genes (JAK3, STAT5a, STAT5b). Enzyme linked immune sorbent assay (ELISA) was used to detect interleukin-2 (IL-2) concentration in subject′s plasma. Measurement data among the three groups that was in accordance with normal distribution was analyzed by one-way analysis of variance, pairwise comparisons using LSD, non-normal distribution data was analyzed by Mann-Whitney test or Kruskal-Wallis H test, and correlation analysis between variables was analyzed using Spearman correlation analysis. Results:① The mRNA expression levels of JAK3, STAT5a and STAT5b were significantly different among the three groups ( F=50.13, P<0.01; F=7.573, P=0.000 7; F=12.14, P<0.01), of which the JAK3 mRNA expression level in the HC group [(606±65)×10 -4] was significantly higher than that in the AG group [(103±13)×10 -4] and IG group [(114±24)×10 -4], and the difference was statistically significant (both P<0.01), while the STAT5a mRNA expression level in the AG group [(89±9)×10 -4] was significantly higher than that in the IG group [(59±4)×10 -4] and HC group [(61±4)×10 -4], and the difference was statistically significant ( P=0.002, P=0.003 9), and the expression level of STAT5b mRNA in HC group [(60±5)×10 -4] was significantly lower than that in AG group [(95±7)×10 -4] and IG group [(98±7)×10 -4], and the difference was statistically significant ( P=0.000 2, P<0.000 1). ② The difference of IL-2 concentration in plasma among the three groups was statistically significant ( F=22.87, P<0.01), and the serum IL-2 concentration in the AG group [(87.9±8.4) pg/ml] was significantly higher than that in the IG group [(32±4) pg/ml] and HC group [(44±4) pg/ml], and the difference was statistically significant (both P<0.01). ③ Spearman correlation analysis showed that the mRNA expression of STAT5a and STAT5b was positively correlated with the absolute value of neutrophils in patients with gout ( r=0.282, P<0.05; r=0.257, P<0.05). Conclusion:The IL-2/JAK3/STAT5 signaling pathway is involved in the occurrence and development of gout, suggesting that this pathway may play a key role in the pathogenesis of gout.

Are stem cell marker expression and cd133 analysis relevant to differentiate colorectal cancer? / A expressão de marcadores de células-tronco e a análise do cd133 são relevantes na diferenciação do câncer colorretal?

ABSTRACT Background: CD133 and AXL have been described as cancer stem cell markers, and c-MYC as a key regulatory cellular mechanism in colorectal cancer (CRC). Aim: Evaluate the prognostic role of the biomarkers CD133, AXL and c-MYC and their association with clinicopathologic characteristics in colorectal adenocarcinomas and adenomas. Methods: A total of 156 patients with UICC stage I-IV adenocarcinomas (n=122) and adenomas (n=34) were analyzed. Tissue microarrays (TMA) from primary tumors and polyps for CD133, c-MYC and AXL expression were performed and analyzed for their significance with clinicopathologic characteristics. Results: Poorly differentiated adenocarcinomas and disease progression were independent risk factors for poor overall survival. The median overall survival time was 30 months. Positive CD133 expression (35.9% of all cases), particularly of right-sided CRCs (44.8% of the CD133+ cases), was negatively correlated with death in the univariate analysis, which did not reach significance in the multivariate analysis. c-MYC (15.4% of all cases) was predominantly expressed in advanced-stage patients with distant (non-pulmonary/non-hepatic) metastasis. AXL expression was found only occasionally, and predominantly dominated in adenomas, with less penetrance in high-grade dysplasia. Conclusions: CD133 expression was not associated with inferior overall survival in CRC. While AXL showed inconclusive results, c-MYC expression in primary CRCs was associated with distant metastasis.

RESUMO Racional: CD133 e AXL são descritos na literatura como marcadores de células-tronco tumorais, e c-MYC cumpre papel chave como mecanismo de regulação celular no câncer colorretal (CCR). Objetivo: Avaliar o papel prognóstico dos biomarcadores CD133, AXL e c-MYC e sua associação com características clinicopatológicas de adenocarcinomas e adenomas colorretais. Métodos: Um total de 156 pacientes com adenocarcinomas de estádio UICC I-IV (n=122) e adenomas (n=34) colorretais foram avaliados. Microarranjos teciduais (TMA) dos tumores primários e adenomas foram realizados em busca de expressão de CD133, c-MYC e AXL, com posterior análise de relação significativa com características clinicopatológicas. Resultados: Adenocarcinomas pobremente diferenciados e progressão de doença foram fatores de risco independentes para má sobrevida global. A taxa mediana de sobrevida global foi de 30 meses. Expressão positiva de CD133 (35,9% dos casos), particularmente em cânceres de cólon direito (44,8% dos casos CD133+), correlacionou-se negativamente com óbito na análise univariada, sem significância estatística na análise multivariada. c-MYC (15,4% dos casos) teve predomínio de expressão em pacientes com estádio avançado com metástases distantes (não-pulmonares/não-hepáticas). Expressão de AXL foi pouco encontrada, com predomínio em adenomas, com menor penetrância em displasia de alto grau. Conclusão: Expressão de CD133 não se associou com sobrevida global inferior em CCR. Enquanto AXL demonstrou resultados inconclusivos, expressão de c-MYC em tumores primários se associou-se à metástases à distância.

Are stem cell marker expression and cd133 analysis relevant to differentiate colorectal cancer? / A expressão de marcadores de células-tronco e a análise do cd133 são relevantes na diferenciação do câncer colorretal?

ABSTRACT Background: CD133 and AXL have been described as cancer stem cell markers, and c-MYC as a key regulatory cellular mechanism in colorectal cancer (CRC). Aim: Evaluate the prognostic role of the biomarkers CD133, AXL and c-MYC and their association with clinicopathologic characteristics in colorectal adenocarcinomas and adenomas. Methods: A total of 156 patients with UICC stage I-IV adenocarcinomas (n=122) and adenomas (n=34) were analyzed. Tissue microarrays (TMA) from primary tumors and polyps for CD133, c-MYC and AXL expression were performed and analyzed for their significance with clinicopathologic characteristics. Results: Poorly differentiated adenocarcinomas and disease progression were independent risk factors for poor overall survival. The median overall survival time was 30 months. Positive CD133 expression (35.9% of all cases), particularly of right-sided CRCs (44.8% of the CD133+ cases), was negatively correlated with death in the univariate analysis, which did not reach significance in the multivariate analysis. c-MYC (15.4% of all cases) was predominantly expressed in advanced-stage patients with distant (non-pulmonary/non-hepatic) metastasis. AXL expression was found only occasionally, and predominantly dominated in adenomas, with less penetrance in high-grade dysplasia. Conclusions: CD133 expression was not associated with inferior overall survival in CRC. While AXL showed inconclusive results, c-MYC expression in primary CRCs was associated with distant metastasis.

RESUMO Racional: CD133 e AXL são descritos na literatura como marcadores de células-tronco tumorais, e c-MYC cumpre papel chave como mecanismo de regulação celular no câncer colorretal (CCR). Objetivo: Avaliar o papel prognóstico dos biomarcadores CD133, AXL e c-MYC e sua associação com características clinicopatológicas de adenocarcinomas e adenomas colorretais. Métodos: Um total de 156 pacientes com adenocarcinomas de estádio UICC I-IV (n=122) e adenomas (n=34) colorretais foram avaliados. Microarranjos teciduais (TMA) dos tumores primários e adenomas foram realizados em busca de expressão de CD133, c-MYC e AXL, com posterior análise de relação significativa com características clinicopatológicas. Resultados: Adenocarcinomas pobremente diferenciados e progressão de doença foram fatores de risco independentes para má sobrevida global. A taxa mediana de sobrevida global foi de 30 meses. Expressão positiva de CD133 (35,9% dos casos), particularmente em cânceres de cólon direito (44,8% dos casos CD133+), correlacionou-se negativamente com óbito na análise univariada, sem significância estatística na análise multivariada. c-MYC (15,4% dos casos) teve predomínio de expressão em pacientes com estádio avançado com metástases distantes (não-pulmonares/não-hepáticas). Expressão de AXL foi pouco encontrada, com predomínio em adenomas, com menor penetrância em displasia de alto grau. Conclusão: Expressão de CD133 não se associou com sobrevida global inferior em CCR. Enquanto AXL demonstrou resultados inconclusivos, expressão de c-MYC em tumores primários se associou-se à metástases à distância.

Effect of Qingzao Jiufei Tang on JAK2/STAT3 Signaling Pathway and Expression of Downstream Apoptosis-related Proteins in Lung Cancer / 中国实验方剂学杂志

Objective::To observe the effect of Qingzao Jiufei Tang on apoptosis of lung cancer, Janus protein tyrosine kinase 2/signal transducers and transcriptional activator protein 3 (JAK2/STAT3) signaling pathway, as well as the expressions of downstream apoptosis-related proteins Bcl-2-associated X (Bax) and Cyclin D1. Method::Totally 50 male C57BL/6J mice were randomly divided into five groups: chemotherapy group (CTX), model group, high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and low-dose Qingzao Jiufei Tang group, with 10 mice in each group. The model of lung cancer was established by injecting Lewis lung cancer cells into the right axillary of mice. High-dose, middle-dose and low-dose Qingzao Jiufei Tang groups were orally given drugs (11, 5.5, 2.75 g·kg-1·d-1) two weeks before the modeling. Chemotherapy group was administered intraperitoneally at a dose of 50 mg·kg-1·(2 d)-1, while model group was administered intragastrically with the equal volume of normal saline. After inoculation, the mice in each group were continued to be administered. Two weeks later, the mice in each group were killed, and the tumors were collected. Then the JAK2 protein phosphorylation level was detected by immunohistochemistry (IHC). STAT3, Bax and Cyclin D1 protein expression levels were detected by Western blot, and apoptosis of lung cancer cells was observed by transmission electron microscopy. Result::Compared with model group, the phosphorylation levels of JAK2 and STAT3 protein in lung cancer cells were significantly decreased, the expression of Bax protein was significantly increased, and the expression of Cyclin D1 protein was significantly decreased in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and chemotherapy group, with statistically significant differences (P<0.05, P<0.01). The results of transmission electron microscopy showed significant apoptotic phenomena in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group, low-dose Qingzao Jiufei Tang group and chemotherapy group compared with the model group. Conclusion::Qingzao Jiufei Tang had an obvious effect in promoting the apoptosis of lung cancer cells. Its mechanism may be related to the inhibition of the phosphorylation of JAK2 and STAT3 protein, the promotion of its downstream Bax protein expression and the inhibition of its downstream Cyclin D1 protein expression.

Preliminary screening of chemotherapy-resistant genes in esophageal cancer and its clinical significance / 解放军医学杂志

Objective To initially screen the genes associated with chemotherapeutic resistance in esophageal cancer cells, verify the correlation of the genes to the poor prognosis of patients with esophageal cancer, and predict the possible regulatory mechanism of esophageal cancer resistance. Methods The drug sensitivity data of esophageal cancer cell lines were analyzed from GDSC database to screen the cell lines that were relatively sensitive or resistant to both cisplatin and docetaxel. In order to obtain differentially expressed genes, the transcriptome data of the two groups of cell lines were analyzed by the edgeR package according to the following screening criteria: the log2 (fold change) more than –1 or less than1 and P value <0.05. The enrichment cluster analysis of GO biological process was performed in the relatively highly expressed genes of drug-resistant group to identify possible signaling pathways related with drug resistance, and the target genes related to chemotherapeutic resistance were identified based on previous studies. The associations between the expression level of target gene and the clinical pathological features and prognosis of patients were verified in the tissue transcriptome data of esophageal cancer patients. Finally, the proteins interacting with the target gene encoded protein were predicted online using the STRING database, and its possible mechanism of action was analyzed. Results Five cell lines with resistance to both cisplatin and docetaxel and 5 sensitive cell lines were obtained. According to the transcriptome data of the two groups of cell lines, 1097 differentially expressed genes were finally obtained, including 532 highly expressed and 565 low expressed genes in the drug resistant group. The results of GO enrichment analysis for the highly expressed genes indicated that the receptor protein tyrosine kinase pathway was obviously enriched. The expression level of FGR involved in this pathway was significantly correlated with tumor T stage (P=0.021), clinical stage (P=0.007) and prognosis (P=0.0021) of patients with esophageal cancer. In addition, protein interaction analysis indicated that FGR interacted directly or indirectly with multiple proteins, mainly in the form of kinase. Conclusions The receptor protein tyrosine kinase pathway is the most significant signaling pathway associated with chemotherapeutic resistance in esophageal cancer cells. The expression level of FGR in this signaling pathway is significantly correlated with the pathological stage and prognosis of patients with esophageal cancer. FGR may regulate the drug resistance of esophageal cancer cells by phosphorylating downstream target proteins.

Depigmentation in a Patient with Chronic Myeloid Leukemia during Chemotherapy with Dasatinib / 대한피부과학회지

No abstract available.

ZAP-70 expression is associated with increased CD4 central memory T cells in chronic lymphocytic leukemia: cross-sectional study

ABSTRACT Background: Although chronic lymphocytic leukemia is basically a B cell disease, its pathophysiology and evolution are thought to be significantly influenced by T cells, as these are probably the most important interaction partner of neoplastic B cells, participating in their expansion, differentiation and survival. Chronic lymphocytic leukemia B cells may also drive functional and phenotypic changes of non-malignant T cells. There are few data about the association between memory T cells and prognosis, especially related to ZAP-70, a common reliable surrogate of the gold standard chronic lymphocytic leukemia prognostic markers. Objective: The aim of this study was to investigate whether the expression of ZAP-70 in chronic lymphocytic leukemia patients is associated with abnormal patterns of the distribution of naïve and memory T cells related to crosstalk between these cells. Methods: In this cross-sectional, controlled study, patients with chronic lymphocytic leukemia were compared with healthy blood donors regarding the expression of ZAP-70 and the distribution of naïve and memory T cell subsets in peripheral blood as measured by flow cytometry. Results: ZAP-70 positive patients presented an increased frequency and absolute number of central memory CD4+ T cells, but not CD8+ T cells, compared to ZAP-70 negative patients and age-matched apparently healthy donors. Conclusions: Because central memory CD4+ T cells are located in lymph nodes and express CD40L, we consider that malignant ZAP-70-positive B cells may receive beneficial signals from central memory CD4+ T cells as they accumulate, which could contribute to more aggressive disease.

Synthesis and activity evaluation of novel tyrosine kinase inhibitors / 国际药学研究杂志

Objective To design and synthesize compounds with protein tyrosine kinase(PTK)inhibitory activity with L029 as the lead compound. Methods L029 derivatives were designed and synthesized from L029 by reduction and/or substitution with the 3-dimethylamino-1-propyl,methyl acetate,methyl propionate in its active H and other sites. PTK activity was measured by enzyme-linked immunosorbent assay(ELISA). The inhibitory rate was calculated to screen out the compounds with PTK inhibitory activity. Re-sults Five target compounds were synthesized and their structures were confirmed by 1H NMR and MS. Three compounds T2,T3 and T5 were screened out with strong PTK inhibitory activity. Conclusion The synthetic routes of the target compounds are simple with mild reaction condition,and 3 compounds show strong inhibitory activity by ELISA. These results can provide reference for the further design and synthesis of this kind of molecules.

Effect of protein tyrosine kinase 6 on human airway epithelial barrier dsyfunction induced by TNF-α / 中国免疫学杂志

Objective:To investigate the effect of protein tyrosine kinase 6(PTK6) on TNF-α induced human airway epithelial barrier dysfunction and mechamism.Methods: After cultivating 16HBE cells in vitro,the recombined PTK6 and PTK6 siRNA was respectively transfected into the cells,with empty vector and scramble siRNA as control.The cells were incubated with exogenous TNF-α.Cell viability was detected by MTT assay.Cells TER and permeability were detected.ZO-1 and Occludin mRNA were analyzed by RT-PCR.PTK6,ZO-1,Occludin,p-ERK1/2,p-JNK1/2 and p-p38MAPK protein were assayed by Western blot.Results: TNF-α remarkably decreased ZO-1 and Occludin mRNA and protein and TER;increased cells permeability,as well as p-ERK1/2,p-JNK1/2 and p-p38 protein(P<0.05).Upregulation of PTK6 further decreased ZO-1 and Occludin mRNA and protein and TER,enhanced cells permeability and p-JNK1/2 and p-p38 protein(P<0.05),and downregulated PTK6 ,the changes of these indices were opposite(P<0.05).Whereas upregulation or downregulation of PTK6 had no effect on p-ERK1/2.Conclusion: Downregulation of PTK6 inhibits the phosphorylation of JNK1/2 and p38MAPK,thus improving TNF-α-induced human airway epithelial barrier dysfunction.

Chronic Myeloid Leukemia Following Radioactive Iodine Treatment for Thyroid Cancer: Two Case Reports and a Literature Review / 대한내과학회지

The incidence of thyroid cancer has increased rapidly worldwide, although most patients can survive for a long time without developing symptoms. While most thyroid cancers are treated with thyroidectomy alone, some patients are given additional radioactive iodine (RAI) in the form of 131I to treat thyroid cancer metastasis. RAI is associated with acute and chronic complications. Secondary malignancies are the most important in long-term cancer survivors. While many studies have reported the occurrence of acute myeloid leukemia after high-dose RAI, there are few reports on chronic myeloid leukemia (CML) after low-dose RAI treatment. Moreover, previous cases of CML following thyroid cancer were reported before the tyrosine kinase inhibitor (TKI) era. Here, we describe two cases of CML following thyroid cancer that were successfully treated with second-generation TKIs.

Expression and clinical significance of Dyrk1b in the specimens and cells of cervical lesions / 中华妇产科杂志

Objective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinase1b (Dyrk1b) in the specimens and cells of cervical lesions. Methods (1)All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion(SIL)admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stageⅠand 15 cases of stageⅡ, 12 cases with low-grade squamous intraepithelial lesion(LSIL)and 40 cases with high-grade squamous intraepithelial lesion(HSIL). While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrk1b was detected by immunohistochemistry among the four groups.(2)The expression of Dyrk1b in HeLa and SiHa cells were detected by western blot method and the expression of Dyrk1b protein were also detected after treatment of AZ191 (5, 10 μmol/L) for 48 hours in HeLa and SiHa cells.(3)The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191(2.5, 5, 10, 25, 50, 100 μmol/L)for 48 hours were detected by methyl thiazolyl tetrazolium (MTT) assay.(4)The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZ191 (5, 10μmol/L) for 48 hours. Results (1)The positive rates of Dyrk1b protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%(3/28), 1/12, 42%(17/40)and 71%(53/75), respectively. The expression of Dyrk1b in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis (P0.05). Expression of Dyrk1b was correlated with stromal invasion depth of cervical cancer (P0.05). (2) Dyrk1b protein was expressed in different levels in HeLa and SiHa cells, and the expression of Dyrk1b was decreased gradually as the increased of the concentration of AZ191 in both HeLa and SiHa cells by treatment of AZ191 for 48 hours. (3) Different concentration of AZ191 treated on cervical cancer cells could inhibit the cellular proliferation and induce cell apoptosis in a concentration-dependent manner(P<0.01), concomitant to the decreased cell survival rate. The apoptosis rate of HeLa and SiHa were increased significantly after 10μmol/L AZ191-treatment for 48 hours, but no any difference induced by 5 μmol/L AZ191-treatment compared to control group. Also,there was no any difference between Hela and SiHa cells in either inhibitory effect or apoptosis rate induced by AZ191. Conclusions Dyrk1b is over-expressed in either specimens or cells of cervical cancer. The expression of Dyrk1b protein in cervical lesions is increased as the progression of disease. Dyrk1b inhibitor AZ191 could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner in cervical cancer cells.

Screening and analysis of Chinese herbs with liver channel tropism acting on human liver cancer cell differential expressed protein tyrosine kinase / 中国中西医结合急救杂志

Objective To screen the Chinese drugs with liver channel tropism acting on the characteristic differential expressed protein of human liver cancer cell protein tyrosine kinase (PTKs) system by protein chip technology, and to analyze the difference in different Chinese drugs with liver channel tropism in relation to PTKs regulation. Methods Forty BALB/C nude mice were chosen; a piece of subcutaneous tumor mass was implanted into the left lobe of liver parenchyma to reduplicate the orthotopically implanted tumor models. After modeling for 10 days, the tumorigenicity was confirmed, and all the nude mice models were divided into four groups; different Chinese medicine extracts were injected intra-peritoneally into corresponding treatment groups respectively, and the methods of treatment in the 4 groups were as follows: In liver channel tropism drug pair of Huayu Xiaozhen group, rhizoma sparganii and curcuma zedoary with dosage containing crude drug 4.5 g·kg-1·d-1 was given, in liver channel tropism drug pair of Gongdu Sanjie group, the extract of centipede and scorpion with dosage containing crude drug 0.3 g·kg-1·d-1 was intra-peritoneally injected, in spleen channel tropism drug pair control group, astragalus and rhizoma atractylodis macrocephalae with dosage containing crude drug 6.3 g·kg-1·d-1 was given, and in the model group, equal amount of 0.9% normal saline was intra-peritoneally injected. After the drug treatment for 3 weeks, the mice were sacrificed and the liver cancer tissue was obtained; after its total protein was extracted, protein chip technology was applied to screen the Chinese drug pair with liver channel tropism acting on differential expressed PTKs of human liver cancer cells, and the results were analyzed by cluster analysis.Results After the end of the experiment, there were 6 animals in Huayu Xiaozhen drug pair group, 5 in Gongdu Sanjie drug pair group, 5 in control drug pair group and 7 in model group. The protein chip screening results showed that the adjusting fold greater > 1.50 or < 0.67 was defined as the difference with statistical significance. Compared with model group, there were 42 up-regulated expressions of PTKs in Huayu Xiaozhen drug pair group, including 29 receptor tyrosine kinases (RTKs) and 13 non-receptor protein tyrosine kinases (nrPTKs) of which the erythropoietin having adjusting fold over 5.0 produced liver cell receptor B1 (EphB1), epidermal growth factor receptor (ErbB2, ErbB4) etc. 3 RTKs; there were 7 RTKs with adjusting fold 3.0 - 5.0 including EphA1, EphA3, EphA7, fibroblast growth factor receptor 2-α (FGFR2-α), hepatocyte growth factor receptor (HGFR), macrophage colony-stimulating factor receptor (M-CSFR) and vascular endothelial growth factor receptor 2 (VEGFR2), and 2 nrPTKs with adjusting fold 3.0 - 5.0 were non-receptor tyrosine kinase BMX (BMX) and Janus kinase 1 (JAK1). In the Gongdu sanjie drug pair group, there were 23 up-regulated expressions: 15 RTKs and 8 nrPTKs, and 6 down-regulated expressions; the RTKs with adjusting fold 3.0 - 5.0 were ErbB4, M-CSFR and 1 nrPTKs that was megakaryocyte-associated tyrosine kinase (MATK). In the control drug pair group, there were 28 up-regulated and 8 down-regulated expressions. The results of cluster analysis showed that in Huayu Xiaozhen drug pair group, there were 17 differential expressed PTKs in accord with the screen criteria of which 9 were RTKs [receptor tyrosine kinase-like orphan receptor 2 (ROR2), stem cell factor receptor (SCFR), anaplasia lymphoma kinase (ALK), platelet-derived growth factor receptor β (PDGFR-β), insulin-like growth factor-IR receptor (IGF-IR), ErbB2, ErbB3, EphB1 and EphA2], 1 nrPTKs [fps/fes related tyrosine kinase (FER)] and 7 PTKs 3 RTKs (M-CSFR, FGFR2-α, EphA3) and 4 nrPTKs [acetate kinase 1 (ACK1), bruton tyrosine kinase (Btk), non-receptor tyrosine kinase ABL1 (ABL1) and BMX]. In Gongdu Sanjie drug pair group, there were 7 differential expressed PTKs in accord with the screen criteria 5 RTKs (M-CSFR, FGFR1, ROR2, EphB1, ErbB2) and 2 nrPTKs [src-related kinase lacking C-terminal regulation and N-terminal myristylation sites (SRMS), FER]. Conclusions The drug pair of centipede and scorpion with Gongdu Sanjie action possesses a more effective anti-HCC role than the drug pair of rhizoma sparganii and curcuma zedoary with Huayu Xiaozhen action, the mechanism is possibly via the regulation of PTKs signal pathway. The liver channel tropism drug pair of rhizoma sparganii and curcuma with action of promoting blood circulation and removing blood stasis possibly has an independent anti-HCC effective pathway outside the PTKs signal system.

The tyrosine kinase inhibitors in combination with radiotherapy enhance the inhibition of breast cancer cells and breast cancer bearing nude mice / 重庆医学

Objective In order to research the effect of tyrosine kinase inhibitor in combination with radiotherapy on the inhibi‐tion of breast cancer cells and breast cancer bearing nude mice .Methods Methyl thiazolyl tetrazolium (MTT) assay was used to e‐valuate the inhibition of different concentrations of TKI on breast cancer cells ,the breast cancer cells was divided into three groups :the TKI treatment group ,the cells in the control group (no the TKI processing) and the control group (non‐TKI and X‐ray irradia‐tion group) .The sensitivity of the cells in each group to X‐ray was compared by colony formation assay .MCF7 cells were xenograf‐ted in athymic nude mice to establish the animal model ,which was used to evaluate the effect of anti‐cancer .Results Colony form‐ing test showed that the separated use of any concentrations of the TKI inhibitor could inhibit the breast cells ,and the cell viability was significantly reduced;TKI combined with X‐ray irradiation could significantly increase the sensitivity of breast cancer cells to radiation ,and the difference was statistically significant(P<0 .05) .Compared to TKI inhibitor or X‐ray irradiation alone ,the combi‐nation of TKI inhibitor with X‐ray irradiation could inhibit the growth of tumor effectively .Conclusion The TKI inhibitor in com‐bination with radiotherapy can effectively inhibit the growth of breast cancer cells ,which provides a new theoretical basis for the im‐plementation of the clinical breast cancer radio sensitization .

Frequency and Clinicohematologic Characteristics of MPL W515 Mutations in Patients with Myeloproliferative Neoplasms

BACKGROUND: Recently, myeloproliferative leukemia (MPL) W515 mutations have been reported to be molecular markers for myeloproliferative neoplasms (MPNs). We studied the association between MPL W515 mutations and the clinico-hematological features of patients with MPNs. METHODS: Our study included 154 consecutive patients diagnosed with MPNs (31 had polycythemia vera [PV]; 106, essential thrombocythemia [ET]; and 17, primary myelofibrosis [PMF]). MPL W515 mutations were detected by real-time PCR and direct sequencing methods. RESULTS: The MPL W515L mutation was found in 4 patients and the MPL W515A mutation was detected in 1 patient. These 5 patients were diagnosed with JAK2 V617F-negative ET, and they accounted for 12.5% of patients with JAK2 V617F-negative ET. The patients with MPL W515-positive ET showed significantly lower hemoglobin levels and WBC counts than did patients with MPL W515-negative ET or JAK2 V617F-positive ET. CONCLUSIONS: MPL W515 mutation is a useful diagnostic marker for JAK2 V617F-negative MPNs and it is associated with specific hematologic characteristics such as lower hemoglobin levels and WBC counts.

Association Analysis of TEC Polymorphisms with Aspirin-Exacerbated Respiratory Disease in a Korean Population

The tyrosine-protein kinase Tec (TEC) is a member of non-receptor tyrosine kinases and has critical roles in cell signaling transmission, calcium mobilization, gene expression, and transformation. TEC is also involved in various immune responses, such as mast cell activation. Therefore, we hypothesized that TEC polymorphisms might be involved in aspirin-exacerbated respiratory disease (AERD) pathogenesis. We genotyped 38 TEC single nucleotide polymorphisms in a total of 592 subjects, which comprised 163 AERD cases and 429 aspirin-tolerant asthma controls. Logistic regression analysis was performed to examine the associations between TEC polymorphisms and the risk of AERD in a Korean population. The results revealed that TEC polymorphisms and major haplotypes were not associated with the risk of AERD. In another regression analysis for the fall rate of forced expiratory volume in 1 second (FEV1) by aspirin provocation, two variations (rs7664091 and rs12500534) and one haplotype (TEC_BL2_ht4) showed nominal associations with FEV1 decline (p = 0.03-0.04). However, the association signals were not retained after performing corrections for multiple testing. Despite TEC playing an important role in immune responses, the results from the present study suggest that TEC polymorphisms do not affect AERD susceptibility. Findings from the present study might contribute to the genetic etiology of AERD pathogenesis.

Clinical trials of receptor protein-tyrosine kinase inhabitors in malignant glioma / 国际肿瘤学杂志

The occurrence and development of malignant glioma are closely related to abnormal overexpression and activation of receptor tyrosine kinase (RTK) signal transduction pathways.Targeted therapeutic drugs such as RTK inhibitors,RTK downstream signaling pathway inhibitors and multi-target inhibitors can targeting treat malignant glioma at molecular level,some of which have been investigated in clinical trials and achieved good therapeutic effects.

The expression of proline-rich tyrosine kinase and its role in systemic lupus erythematosus / 中华风湿病学杂志

Objective To explore the possible role of proline-rich tyrosine kinase (Pyk2) in the pathogenesis of systemic lupus erythematosus (SLE).Methods The expression of Pyk2 in the peripheral blood mononuclear cells (PBMCs) of 50 patients with SLE and 36 healthy controls were tested with RT-PCR assay.The activation of Pyk2 was inhibited using specific inhibitor Pyk2 (TyrA9).Semi-quantitative PCR methodwas used to detect the Blys expression of PBMCs.One-way ANOVA and Pearson's correlation analysis were used for statistical analysis.Results The Pyk2 expression level (28.31 ±0.91) of SLE patients was significantly higher than that in healthy controls (33.69±0.04),the difference was statistically significant (P＜0.05).The activation of Pyk2 was stimulated and the expression levels of Blys in the PBMCs of patients with SLE was elevated.By inhibiting the activation of Pyk2,the BLyS expression levels decreased significantly.Conclusion Pyk2 may be involved in the abnormal activation of lymphocytes which lead to the pathogenesis of SLE.Pyk2 expression is associated with SLE disease activity,disease aggravation,and the Pyk2 expression levels is also increased significantly.In addition,the expression level of Pyk2 is higher in patients with renal involvement than those patients with other organ involvement.

Detecção de mutações no gene KIT em leucemia mieloide aguda / The detection of KIT mutations in acute myeloid leukemia

OBJETIVO: Descrever a metodologia para detecção de mutações nos éxons 8 e 17 do gene KIT em pacientes portadores de leucemia mieloide aguda, para implementação desse teste no laboratório clínico do Hospital Israelita Albert Einstein. MÉTODOS: Extração do DNA genômico de 54 amostras de sangue periférico ou medula óssea de pacientes com leucemia mieloide aguda para amplificação, por reação em cadeia da polimerase, sequenciamento e análise de fragmentos. RESULTADOS: Dentre as amostras analisadas, quatro apresentaram mutação no éxon 8, duas no éxon 17 e uma amostra apresentou mutação nos dois éxons. CONCLUSÃO: A pesquisa de mutação nos éxons 8 e 17 do gene KIT foi padronizada com sucesso e o teste está em processo de inclusão no menu de exames do laboratório clínico do Hospital Israelita Albert Einstein.

OBJECTIVE: This study describes a new method used in the clinical laboratory at Hospital Israelita Albert Einstein to detect mutations in exons 8 and 17 of the KIT gene in patients with acute myeloid leukemia. METHODS: Genomic DNA extraction was performed on 54 samples of peripheral blood or bone marrow from patients with acute myeloid leukemia. The extracted DNA was amplified by polymerase chain reaction and sequenced, and the fragments were analyzed. RESULTS: Within the analyzed samples, we detected four mutations in exon 8, two mutations in exon 17, and mutations or a double mutation in one sample. CONCLUSION: The tests detecting mutations in exon 8 and 17 on the KIT gene were successfully standardized. The test is now included among the routine diagnostics employed for patients at Hospital Israelita Albert Einstein clinical laboratory.