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Langerhans cell histiocytosis (LCH), a disorder of antigen-presenting cells, is the commonest disorder of the mononuclear phagocytic system. Diagnosis is always challenging due to heterogeneous clinical presentation. However, with the evolution and better understanding of its biology, many of these children are being diagnosed early and offered appropriate therapy. Despite these advances, in developing countries, an early diagnosis is still challenging due to resource constraints for specialized tests. As a result, many patients succumb to their disease. Autopsy data on LCH is notably lacking in the literature. We sought to analyze the clinical (including mutational) and morphologic features at autopsy in six proven cases of LCH. This study includes a detailed clinico-pathological and mutational analysis of 6 proven cases of LCH. Presence of BRAF V600E mutation was assessed by both Real Time PCR and Sanger sequencing. A varied spectrum of organ involvement was noted with some rare and novel morphological findings, like nodular bronchiolocentric infiltration of LCH cells, lymphovascular emboli of LCH cells, and paucity of eosinophils within the infiltrate; these features have not been described earlier. Surprisingly, all cases were negative for BRAF V600E mutation on both RQ-PCR and Sanger sequencing. The present study is perhaps the first autopsy series on LCH. This extensive autopsy analysis represents a correlation of pathological features with clinical symptoms which provides clues for a timely diagnosis and appropriate therapeutic intervention. Also, our findings hint at the low frequency of BRAF V600E mutation in our LCH patients.
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Humans , Male , Infant , Child, Preschool , Histiocytosis, Langerhans-Cell/pathology , Autopsy , Proto-Oncogene Proteins c-abl , Mitogen-Activated Protein Kinase Kinases , Early DiagnosisABSTRACT
Objective To explore the effect of proliferation and apoptosis,and research its mechanism associated with RAS/extracellular signal regulated kinase/mitogen-activated protein kinase (RAS/ ERK/MAPK) in Fms-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD) mutation acute lymphocytic leukemia cell line MV4-11 cells treated by triptolide (TP).Methods MV4-11 cells were respectively treated by triptolide with diverse concentrations and different times.The proliferation inhibition rate was measured by methyl thiazolyl tetrazolium,the apoptosis rate was detected by flow cytometry,the mRNA expressions of FLT3,RAS,ERK,forkhead box protein M1 (FOXM1),and c-Myc were analyzed by realtime fluorescent quantitative polymerase chain reaction (PCR).Results The proliferation inhibition rate markedly increased in a dose-time dependent manner after MV4-11 cells were treated by TP.After cells were treated with 10,and 20 nmol/L TP,respectively,the apoptosis rates at 48 h were (17.30 ± 0.56) ％,(35.77 ±0.55)％,and those at 72 h were (49.83 ±0.45)％,(68.90 ±0.75)％ correspondingly.PCR data showed that the mRNA level of FLT3 mRNA decreased obviously,and that of RAS,ERK,FOXM1,and c-Myc also decreased.Conclusions Triptolide could significantly inhibit the MV4-11 cells proliferation and induce cell apoptosis,and its mechanism might be through inhibiting the expression of related genes on RAS/ERK/MAPK signaling pathway.
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Tumor progression locus 2 (Tpl2) is a crucial three-stage kinase of the mitogen-activited protein kinase (MAPK) family,which plays an important role in MAPK pathway and other signaling pathways.In recent years,a large number of studies have found that aberrant expressing Tpl2 is involved in tumorigenesis and development of various cancers,and is expected to serve as a new biomarker and therapeutic target.Therefore,to reveal the mechanism of Tpl2 will provide a feasible theoretical basis and potential interventional target for the diagnosis and treatment of cancers.
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Objective To study the effect of mitogen-activated protein kinas/extracellular signalregulated kinase (MAPK/ERK) signaling pathway on cell proliferation modulated by Sonic Hedgehog (Shh) signaling in fibroblast-like synoviocytes (FLS) isolated from patients with active rheumatoid arthritis (RA).Methods The synovial tissue were collected by the synovial arthroscopic debridement or arthroscopic synovectomy of RA patients with active disease activity [disease activity score(DAS)28 ≥3.2].The RA-FLS were primarily cultured by the explanted culture,and then were treated with Shh agonist purmorphamine,inhibitor cyclopamine or MAPK/ERK signaling pathway inhibitor U0126,respectively.Western blotting was used to examine the phosphorylation level of ERK 1/2 (p-ERK1/2),which was the critical protein of MAPK/ERK signaling.The cell proliferation activity was detected using cell proliferation and cytotoxicity kit-8 (CCK8),and the cell proliferation rate was detected using a flow cytometry.Analysis of variance and Kruskal-Wallis H(K) test were used for statistical analysis.Results Compared with the control group,purmorphamine transiently increased p-ERK1/2 protein at the concentration of 1 μmol/L,and the peak activations of p-ERK1/2 took place at 15 min (P＜0.01).Cyclopamine and U0126 decreased the expression ofp-ERK1/2 protein (P＜0.01).After the RA-FLS treated with purmorphmine(1 μmol/L)for 48 hours,the cell proliferation activity was (114±4)％ and the percentage of S phase cells was (8.39±0.60)％,which was significantly higher than those of the control group (100±0)％ (P＜0.01) and (3.29±0.69)％ (P＜0.01).After treated with cyclopamine (10 μmol/L) for 48 hours,the cell proliferation activity of RA-FLS was (89±1)％ (P＜0.05) and the percentage of S phase cells was (1.53±0.22)％ (P＜0.05).When co-treated with purmorphamine (1 μmol/L) and U0126 (10 μmol/L),the cell proliferative activity was (89±2)％ (P＜0.05) and the percentage of S phase cells was(1.07±0.25)％(P＜ 0.05).Conclusion Shh may promote proliferation of RA-FLS via modulating MAPK/ERK signaling,which in turn contributes to hyperplasia of synovium and ultimately leading to RA.
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Melanoma is one of the most aggressive cancers in the world and is responsible for the majority of skin cancer deaths. Recent advances in the field of immunotherapy using active, adoptive, and antigen-specific therapeutic approaches, have generated the expectation that these technologies have the potential to improve the treatment of advanced malignancies, including melanoma. Treatment options for metastatic melanoma patients have been dramatically improved by the FDA approval of new therapeutic agents including vemurafenib, dabrafenib, and sorafenib. These kinase inhibitors have the potential to work in tandem with MEK, PI3K/AKT, and mTOR to inhibit the activity of melanoma inducing BRAF mutations. This review summarizes the effects of the new therapeutic agents against melanoma and the underlying biology of these BRAF inhibitors.
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Humans , Biology , Immunotherapy , Melanoma , Mitogen-Activated Protein Kinase Kinases , Phosphotransferases , Skin NeoplasmsABSTRACT
Melanoma is one of the most aggressive cancers in the world and is responsible for the majority of skin cancer deaths. Recent advances in the field of immunotherapy using active, adoptive, and antigen-specific therapeutic approaches, have generated the expectation that these technologies have the potential to improve the treatment of advanced malignancies, including melanoma. Treatment options for metastatic melanoma patients have been dramatically improved by the FDA approval of new therapeutic agents including vemurafenib, dabrafenib, and sorafenib. These kinase inhibitors have the potential to work in tandem with MEK, PI3K/AKT, and mTOR to inhibit the activity of melanoma inducing BRAF mutations. This review summarizes the effects of the new therapeutic agents against melanoma and the underlying biology of these BRAF inhibitors.
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Humans , Biology , Immunotherapy , Melanoma , Mitogen-Activated Protein Kinase Kinases , Phosphotransferases , Skin NeoplasmsABSTRACT
Objective To investigate the activation of β-sheet breaker peptide H102 on ERK signal transduction pathway in brain of PAP double transgenic mice. Methods PAP double transgenic mice were randomly divided into model group and H102 treatment group (n=10 for each group). A group of C57BL/6J mice with the same genetic background was served as controls. H102 (5.8 mg/kg) 5 μL was infused by intranasal administration to mice in H102 treatment group, and equal volume of blank solution of H102 (chitosan, BSA) was given to mice in control group and model group. The ability of spatial reference memory was tested by Morris water maze after 30 days of treatment. Then immunohistochemistry tests and Western blot technique were used to detect the content of RAS, P-MEK and P-ERK proteins in mouse brain. Results (1) The ability of learning and memory was significantly lower in model group than that of control group. The ability of learning and memory was significantly improved in treatment group than that in model group (P<0.05). (2) The contents of RAS, P-MEK and P-ERK in mouse brain were significantly lower in model group than those of control group, and these protein ex-pressions were significantly increased in treatment group than those in model group (P<0.01). Conclusion β-sheet break-er peptide H102 can activate ERK signal transduction pathway in brain of PAP double transgenic mice, increase PAS, P-MEK and P-ERK levels in nerve cells, and improve the ability of learning and memory in PAP mice.
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Objective To investigate the expression of Hedgehog signaling pathway members,Shh,Gli1,Sufu and TAK1 as well as phosphorylation-TAK1 (p-TAK1) protein in human pancreatic carcinoma tissues and to explore its relationship between the expression of proteins and the clinicopathologic parameters.Methods The expressions of Shh,Glil,Sufu,TAK1,p-TAK1 proteins in 38 samples of pancreatic cancer tissues and pairing adjacent normal pancreatic tissues were detected by immunohistochemical method.The relationship among the proteins and the relationship between the expression of the proteins and the clinicopathologic parameters were examined.Results The positive expression rates of Shh,Glil,Sufu,TAK1,p-TAK1 protein in human pancreatic carcinoma were 86.8％ (33/38),52.6％ (22/38),68.4％(26/38),55.3％ (21/38),52.6％ (20/38),but they were not expressed in adjacent normal pancreatic tissues.Gli1 expression was positively related to distant metastasis and clinical staging (r =0.524,0.361,P ＜0.05),Sufu expression was positively related to patient's gender (r =-0.378,P ＜0.05),TAK1 expression was positively related to clinical staging (r =0.468,P ＜ 0.05),p-TAK1 expression was positively related to clinical staging and distant metastasis (r =0.418,0.361,P ＜ 0.05).Gli1 expression was positively related to TAK1 and p-TAK1 expression in pancreatic cancer tissues (P ＜ 0.05).Conclusions Hedgehog signaling pathway and p-TAK1 may play a role in the pathogenesis of pancreatic cancer,and the two pathways may interact with each other.
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Objective To evaluate the effect of resveratrol on the growth of an established A431 xenogratt tumor in nude mice.Methods The model of human skin squamous cell carcinoma was established by inoculating A431 cells in log-phase growth into the left axillary fossa of Balb/c (nu/nu) nude mice.After 7-8 days,60 mice bearing human A431 skin squamous cell carcinoma xenografts were randomly and equally divided into 6 groups:blank control group receiving no treatment,negative control group treated with intraperitoneal sodium chloride physiological solution,positive control group treated with intraperitoneal cyclophosphamide,high-,medium-and low-dose resveratrol groups treated with intraperitoneal resveratrol of 40,20 and 10 μg per gram body weight per day,respectively.Tumor size was measured at a 4-day interval during the treatment course.After 14-day treatment,the mice were sacrificed.Xenograft tumors were removed from these mice and subjected to weight measurement,pathological examination by hematoxylin and eosin (HE) staining and apoptosis detection by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Western blot was conducted to quantify the protein expression of apoptosis-related factors,including phosphorylated extracellular signal-regulated protein kinase (p-ERK),p53 and caspase 3.Data were processed by SPSS 13.0 software,and statistical analysis was carried out by analysis of variance and Pearson correlation analysis.Results By the end of treatment,the xenograft tumor volume was (1153.56 ± 255.41) mm3,(1001.69 ± 115.08) mm3,(1206.80 ± 175.88) mm3,(1342.28 ± 211.12) mm3,(1642.34 ± 225.85) mm3 and (1564.32 ± 156.49) mm3,and the weight was (1.84 ±0.30) g,(1.72 ± 0.39) g,(1.96 ± 0.40) g,(2.67 ± 0.73) g,(3.16 ± 0.52) g,and (3.33 ± 0.59) g,respectively in the positive control group,high-,medium-and low-dose resveratrol group,negative control group and blank control group.Significant differences were observed in the xenograft tumor volume (F =16.00,P ＜ 0.05) and weight (F =19.15,P ＜ 0.05) among the 6 groups.According to the tumor weight,the growth of tumor was inhibited by 45.57％,37.97％ and 15.51％ respectively in the high-,medium-and low-dose resveratrol groups.Increased apoptotic index was observed in the positive control group,high-,medium-and low-dose resveratrol groups compared with the negative control group and blank control group (36.79 ± 8.86,33.15 ± 6.00,18.09 ±3.92 and 10.53 ± 4.20 vs.3.87 ± 1.63 and 2.73 ± 1.61,F =93.26,P ＜ 0.05).Analysis of variance showed that the protein expressions of p-ERK,p53 and caspase 3 were all higher in the three resveratrol groups than in the negative control group and blank control group (F =6.65,6.78,11.56,respectively,all P ＜ 0.05).The protein expression of p53 was statistically correlated with p-ERK (r =0.68,P ＜ 0.05) and caspase 3 (r =0.56,P ＜0.05).Conclusions Resveratrol shows an inhibitory effect on the growth of human A431 skin squamous cell carcinoma xenografts in nude mice,likely by increasing p53 expression and inducing tumor cell apoptosis via the activation of MAPK/ERK pathway.
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Objective To investigate the changes in the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) in the distal cerebrospinal fluid contacting neurons (CSF-CNs) in rats with neuropathic pain.Methods Forty-eight male adult SD rats aged 2-3 months weighing 230-270 g were randomly divided into 2 groups (n =24 each): sham operation group (group S) and chronic constrictive injury group (group CCI).Neuropathic pain was induced by CCI.Sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 silk thread.Thermal withdrawal latency (TWL) was measured with radiation heat stimulus before (baseline) and 1,3,5,7,14 d after CCI in 4 animals at eachtime point in each group.The animals were then sacrificed.30％ cholera toxin subunit B with horse radish peroxidase 3 μl was injected into the lateral ventricle at 48 h before the animals were sacrificed at each time point.The brain tissue in the region of the midbrain aqueduct was taken.The number of distal CSF-CN expressing p-p38MAPK was calculated.p-p38MAPK was detected by histochemical staining.Results The TWL was significantly shortened at 1-14 d after CCI in group CCI as compared with group S.The number of distal CSF-CNs expressing p-p38MAPK was significantly increased at 5-14 d after CCI in group CCI as compared with group S.Conclusion The distal CSF-CNs may be involved in the neuropathic pain induced by CCI through p38MAPK signaling pathway.p38MAPK may contribute only to the maintenance but not development of neuropathic pain.
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Objective: To investigate the association between the single nucleotide polymorphism (SNP) of mitogen-activated protein kinase kinase 4 (MKK 4) promoter and the risk of lung cancer development in Southern Chinese population. Methods: A hospital-based case-control study including 800 cases of lung cancer and 900 healthy controls was conducted. TaqMan assay was used to test the SNP of rs3826392 (-1304T>G) in MKK 4 promoter, and the software SAS 9.13 was used to analyze the association of MKK4 polymorphism and the susceptibility of lung cancer. Results: The observed genotype frequency of -1304T>G in MKK4 promoter was appropriate for Hardy-Weinberg equilibrium in the healthy controls (P = 0.149). The distribution of genotypes was significantly different between the cases of lung cancer and the healthy controls (P=0.001). Compared with homozygous genotype (TT), the risk for lung cancer was decreased by 25% in the carriers of heterozygous genotype (TG) [adjusted odd ratio (OR ) = 0.75, 95% confidence interval (CI) = 0.58-0.97)], and the risk for lung cancer in carriers of GG homozygote was decreased by 45% (adjusted OR = 0.55, 95% CI = 0.33-0.94). There was a significantly decreased trend in the risk for lung cancer along with the increased number of mutation-typic G allele (PtrendG genotype in promoter region of MKK4 gene may contribute to the decreased risk of lung cancer. Copyright© 2011 by TUMOR.
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AIM: To study the expression and the role of ERK1/2 and JNK1/2 of MAPKs pathways in the development of neural tube defects induced by hyperthermia. METHODS: The animal models of golden hamster were produced by hyperthermia. The expression of ERK1/2 and JNK1/2, and levels of their phosphorylation were measured by Western blotting in control group and hyperthermia group. RESULTS: p-ERK1/2 steadily expressed in each control group, and the expression of p-ERK1/2 significantly decreased, which was different from that in the corresponding control group (P<0.05). The activity of p-JNK1/2 increased in hyperthermia group and the amount of p-JNK1/2 increased as compared to control group. The peak appeared at 16 h after exposed to hyperthermia (P<0.05). CONCLUSION: Hyperthermia, which induces a decrease in p-ERK1/2 expression and increases the expression of p-JNK1/2 of MAPKs pathway, results in the unbalance of cell proliferation and apoptosis, and induces neural tube defects.
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Objective To evaluate the role of MLK3-MKK3/6-p38MAPK signal transduction pathway in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. Methods Seventy-eight healthy adult male SD rats weighing 200-250 g were randomly divided into 4 groups: control group (group C, n = 6), ALI group ( n =24), MLK3 inhibitor K252a group (group MK, n = 24) and p38MAPK specific inhibitor SB203580 group (group MS, n = 24). ALI was induced by iv injection of LPS 5 mg/kg via tail vein, while the equal volume of normal saline was given instead in group C. K252a 75 μg/kg and SB203580 10 mg/kg were injected intravenously via tail vein 30 min before LPS administration in group MK and MS respectively. The rats were killed at 1, 3, 6 and 12 h (T1-4) after LPS administration in group ALI, MK and MS (6 rats at each time point) and immediately after normal saline administration in group C. The lungs were removed for microscopic examination. The left lung was lavaged.The broncho-alveolar lavage fluid (BALF) was collected. The concentration of TNF-α in the BALF was determined by ELISA. W/D lung weight ratio was calculated. The expression of p-MLK3, p-MKK3/6 and p-p38MAPK were determined by Western blot. Results The concentration of TNF-α in the BALF, W/D lung weight ratio, and expression of p-MLK3, p-MKK3/6 and p-p38MAPK were significantly higher in the other three groups than in group C (P ＜ 0.01). The parameters mentioned above were significantly lower in group MK, and the concentration of TNF-α in the BALF, W/D lung weight ratio, and p-p38MAPK expression were significantly lower in group MS than in group ALI (P ＜ 0.05). The microscopic examination showed that LPS-induced ALI was less severe in group MK and MS than in group ALI. Conclusion MLK3-MKK3/6-p38MAPK signal transduction pathway plays an important role in LPS-induced ALI in rats.
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Objective: To determine the role of p38 mitogen-activated protein kinase (MAP) kinase signal transduction pathways in epigallocatechin-3-gallate (EGCG)-induced apoptosis in human gastric cancer MGC803 cells. Methods: The viability of MGC803 cells was measured by MTT assay. Apoptosis of MGC803 cells was observed by AO/EB fluorescence microscopy and detected by flow cytometry with PI staining. Expression of p38MAPK and phosphorylated p38 (pp38) MAPK were determined by Western blot analysis. Results: EGCG induced apoptosis of MGC803 cells and apparently increased the activity of pp38MAPK in MGC803 cells. However, after interference with pp38MAPK inhibitor, the inhibitory effect of EGCG on MGC803 cells was significantly weakened. The apoptotic rate of the cells and the activity of pp38MAPK also decreased dramatically. Conclusions: EGCG can induce apoptosis of MGC803 cells. The effects could be markedly suppressed by pp38MAPK inhibitor, SB203580. EGCG induces apoptosis of MGC803 cells partly by activating p38 MAPK.
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Objective To investigate the expression of activated ERK (p-ERK) and activated p38 (p-p38) in the keratinocytes of condyloma acuminata (CA) lesions. Methods Fifty cases of HPV 6/11 CA were diagnosed by in situ hybridization. The expression and distribution of p-ERK and p-p38 in CA lesions and 25 normal human skins (foreskins) were detected by immunohistochemistry technique (En Vision). Results ①The results showed that the expression of p-ERK and p-p38 in keratinocytes of CA lesions were significantly higher than those in normal epidermis (P
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Objective To study the effect of TGF-?1 on the activation of ERK MAPK in human bronchial epithelial BEP2D cells. Methods Western blot was employed to examine the time-dependent activation of ERK MAPK by TGF-?1. BEP2D cells were harvested after treatment of human bronchial epithelial cells with 2 ng/ml TGF-?1 for 0, 10, 30, 60, 120, 240 and 480 min, respectively. Fluorescent dye staining and flow cytometry were employed to assess the apoptosis of BEP2D cells treated with vehicle, or with 2ng/ml TGF-?1, or co-treated with 2ng/ml TGF-?1 and 5?M U0126. Proliferation of BEP2D cells treated with vehicle, or with 2ng/ml TGF-?1 or 5?M U0126, or co-treated with 2ng/ml TGF-?1 and 5?M U0126 was assayed with colony-forming test, respectively. Morphological observation was performed to observe the morphological changes in BEP2D cells treated with vehicle, or with 5ng/ml TGF-?1 or 5?M U0126, or co-treated with 5ng/ml TGF-?1 and 5?M U0126, respectively. Results TGF-?1 activated ERK MAPK in BEP2D cell. The maximal activation of ERK MAPK took place at 60min after stimulation with 2ng/ml TGF-?1. TGF-?1 treatment effectively inhibited cell proliferation, and induced their apoptosis and epithelial-mesenchymal transition. Pretreatment with U0126, an inhibitor of ERK MAPK, significantly enhanced the TGF-?1-mediated anti-proliferation and apoptosis effects, and inhibited the effect of epithelial-mesenchymal transition of TGF-?1 in BEP2D cells. Conclusion TGF-?1-induced phosphorylation of ERK MAPK may participate in BEP2D cell proliferation and apoptosis regulation.
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Objective To investigate p38 MAPK signal transduction pathway of cytokine release in co-culture of BEAS-2B cells and eosinophils.Methods Eosinophils in human peripheral blood were isolated using anti-CD16 magnetic micro beads.Co-culture of BEAS-2B cells and eosinophils was taken as experimental model to investigate the inhibitive effect of SB 203580,a selective inhibitor of p38 MAPK,on release of IL-6,IL-8,IP-10 and MIG in culture supernatant.IL-6 was determined by ELISA kit and IL-8,IP-10 and MIG were measured using cytometric bead array(CBA)kit.Results SB 203580 could inhibit IL-6 and IL-8 release from BEAS-2B cells(P