Molecular mechanism of topoisomerase I inhibitor in protecting against experimental acute pancreatitis in mice / 中华胰腺病杂志

Objective:To investigate the protective mechanism of topoisomerase I inhibitor on pancreatic acinar cells and lung during acute pancreatitis (AP) in mice.Methods:Eighteen Balb/C male mice were randomly divided into three groups using random number method: control group, AP group and CPT+ AP group. AP model was established by intraperitoneal injection of caerulein and lipopolysaccharide. CPT+ AP group received intraperitoneal injection of camptothecin (CPT, 50 mg/kg) before AP induction. Mice in control group were intraperitoneal injected with an equal volume of normal saline. The pathological examinations of pancreas and lung tissue were analyzed. The serum levels of amylase and lipase were detected by enzyme linked immunosorbent assay (ELISA) and the mRNA expression of IL-1 and IL-6 were analyzed by reverse transcription-polymerase chain reaction (RT-PCR); the infiltration of CD 45+ cells in pancreas and lung tissue as well as the expression of phosphorylated mixed lineage kinase domain-like protein(MLKL) in pancreas were detected by immunohistochemistry; the apoptosis index of pancreatic cells was analyzed by TUNEL assay. Results:The pathological scores of pancreas and lung tissue, serum levels of amylase and lipase in CPT+ AP group were [(2.30±0.31), (2.29±0.34), (1742.33±183.51)U/L and (46.90±2.17)U/L], which were significantly lower than those in AP group [(5.06±0.88), (3.40±0.09), (2385.33±383.10)U/L and (69.13±9.76)U/L]; the mRNA expression of IL-1 and IL-6 in pancreatic tissue were 95.79±48.11, 255.50±213.32, which were also remarkably lower than those in AP group (212.35±80.61, 1006.80±509.06); the infiltration of CD 45+ inflammatory cells in pancreas and lung were (14.25±5.32, 29.20±4.44)/high power field, which were notably lower than those in AP group (59.83±13.67, 58.25±5.91)/high power field; the apoptosis index of pancreatic cells was significantly higher than that in AP group [(3.64±1.16)% vs (1.92±0.29)%]; the histochemistry score of phosphorylated MLKL protein in pancreatic tissue was significantly lower than that in AP group (1.75±0.20 vs 4.53±1.28), and the differences were statistically significant (all P value <0.05). Conclusions:Topoisomerase I inhibitor could induce the apoptosis of pancreatic acinar cells and inhibit the death mode of necrotic pancreatic acinar cells during AP remodeling, thus reducing pancreatic local injury and AP-associated lung injury.

Autoanticorpos em esclerose sistêmica e sua correlação com as manifestações clínicas da doença em pacientes do Centro-Oeste do Brasil / Autoantibodies in systemic sclerosis and their clinical correlation in patients from a Midwestern region of Brazil

Introdução: a esclerose sistêmica (ES) é uma enfermidade do tecido conjuntivo de caráter autoimune caracterizada pela tríade de injúria vascular, autoimunidade (celular e humoral) e fibrose tecidual. Os autoanticorpos não parecem ser simplesmente epifenômenos, mas sim estarem envolvidos na patogênese da doença. Acredita-se que os autoanticorpos específicos da ES são responsáveis tanto pela amplificação da resposta imune quanto por alvejar os tipos celulares que são relevantes na fisiopatologia da ES. Objetivos: correlacionar o perfil de autoanticorpos específicos (anti-SCL70, ACA, anti-POL3) com as manifestações clínicas e laboratoriais observadas em 46 pacientes com ES da região Centro-Oeste do Brasil. Métodos: pesquisou-se a ocorrência de autoanticorpos específicos em 46 pacientes com diagnóstico de ES e correlacionou-se o tipo de autoanticorpo com as manifestações clínicas e laboratoriais encontradas. Resultados: dentre todos os pacientes avaliados, encontrou-se predomínio feminino (97,8%), idade média de 50,21 anos, cor branca (50%), forma limitada da doença (47,8%), tempo de diagnóstico entre cinco e 10 anos (50%) e tempo de evolução da doença de 9,38 anos. De acordo com o autoanticorpo específico, 24 pacientes apresentavam ACA positivo (52,2%), 15 apresentavam positividade para anti-SCL70 (32,6%) e sete apresentavam anti-POL3 positivo (15,2%). O autoanticorpo anti-SCL70 se correlacionou com a forma difusa da doença, com maior gravidade e atividade da doença, com pior qualidade de vida medida pelo índice HAQ, com maior prevalência de fenômeno de Raynaud objetivo e microcicatrizes de polpas digitais. O ACA se correlacionou com a forma limitada da doença, com o início mais precoce da enfermidade, bem como com maior prevalência de telangiectasias nos pacientes. Já o anti-POL3 se correlacionou com a forma difusa da doença, com maior ocorrência de fenômeno de Raynaud subjetivo e de atrofia muscular. Para as demais variáveis relacionadas ...

Introduction: Systemic sclerosis (SSc) is a connective tissue disease of autoimmune nature characterized by the triad of vascular injury, autoimmunity (cellular and humoral) and tissue fibrosis. Autoantibodies do not seem to be simply epiphenomena, but are involved in disease pathogenesis. It is believed that the SSc-specific autoantibodies are responsible both for amplifying immune response and targeting cell types that are relevant in the pathophysiology of SSc. Objectives: To correlate the profile of the following specific autoantibodies: anti-centromere (ACA), anti-topoisomerase I (topo I) and anti-RNA polymerase III (RNAP III) with clinical and laboratory manifestations were observed in 46 patients with SSc in the Midwest region of Brazil. Methods: The occurrence of specific autoantibodies in 46 patients with SSc was investigated, correlating the type of autoantibody with clinical and laboratory manifestations found. Results: Among all patients evaluated, we found a predominance of females (97.8%), mean age 50.21 years old, Caucasian (50%), limited cutaneous SSc (47.8%), time of diagnosis between 5 and 10 years (50%), and disease duration of 9.38 years. According to the specific autoantibody profile, 24 patients were ACA-positive (52.2%), 15 were positive for anti-topo I (32.6%), and 7 showed positive anti-RNAP III (15.2%). The anti-topo I autoantibody correlated with diffuse scleroderma, with greater disease severity and activity, with worse quality of life measured by the SHAQ index, with a higher prevalence of objective Raynaud's phenomenon and digital pitting scars of fingertips. The ACA correlated with limited scleroderma, with earlier onset of disease, as well as higher prevalence of telangiectasias. The anti-RNAP III correlated with diffuse scleroderma, with a higher occurrence of subjective Raynaud's phenomenon and muscle atrophy. There was no association between the positivity for anti-topo I, ACA and anti-RNAP III antibodies ...

Avances en los mecanismos moleculares del cáncer / Advances in the molecular mechanisms of cancer

In this review, epidemiological, physiological, pathophysiological and pharmacological themes of cancer are dealt. So far, there are over 200types of cancers, which are linked to six key events that collectively lead to the formation of a malignance: self-sufficiency in growth signals, insensitivity to growth inhibitory signals, evasion of apoptosis, unlimited replication potential, sustained angiogenesis and invasion and metastasis. These six capabilities are possibly shared by most human tumors. In2000, there were 10 million new cancer cases and 6 million cancer deaths worldwide. According to estimates by the American Cancer Society, the disease produced approximately 556,000 deaths in 2003, corresponding to 1,500 deaths from cancer every day in America. Annually, in Chile, cancer is responsible for 23 percent of all deaths, constituting the second leading cause of death after cardiovascular diseases. They have identified several risk factors for cancer such as smoking, chronic infections, alcohol consumption, reproductive factors, hormone replacement therapy, dietary habits, sunlight, among others. These factors may cause multiple genetic alterations that involve activation of several oncogenes and the loss of two or more suppressor genes, but not a single change will lead to the formation of a neoplasm. The Knowledge of the molecular differences between normal and malignant cells could be used to target specific pathways and receptors of the latter, thus preventing normal cell death.

Cytotoxic effects of Belotecan in the cervical cancer cell lines / 대한산부인과학회지

OBJECTIVE: In our domestic market, Belotecan (Camptobel(R), chongkeundang, Korea) is newly introduced recently. Belotecan has many advantages of improved water solubility and fewer side effects like severe diarrhea or GI bleeding compare to other camptothecin derivatives. In this study, primary focus is aiming to evaluate the effectiveness of belotecan by providing the cytotoxicity and apoptotic pathway on cervical cancer cells. METHODS: Cervical cancer cell line, HeLa and Caski were used. Belotecan applied on both cell lines and checked whether it has anti tumor effect on cancer cell by using MTT assay. DNA fragmentation and western blot was performed to confirm cellular apoptosis pathway. Also cDNA microarray and RT-PCR were serially carried out in order to identify responsible genes for apoptosis. RESULT: Dose- and time- dependent inhibition of cell proliferation is noted on the Belotecan applied HeLa and CaSki cervical carcinoma cell line by MTT assay. DNA fragmentation assay showed the DNA ladder indicating apopoptosis. Also apoptotic pathway and genes that are related with Belotecan activities are identified. Apoptosis, cell cycle, and drug metabolism related gene, and DNA repair gene were found to be differently regulated by treatment of Belotecan in HeLa cells. Among the DNA repair gene, RT-PCR reconfirmed the increased expression of CIB1(Calcium and intergrin binding 1), APEX1 (APEX nuclease 1) and the decresed expression of EXO1 (Exonuclease 1), WDR33 (WD repeat domain 33), and GADD45A (Growth arrest and DNA-damage-inducible, alpha). CONCLUSION: The first domestically introduced 1st line anti- tumor agent, Belotecan shows its excellent inhibiting action on cervical cancer cell proliferation by apoptotic pathway in this study. Also genetic alterations in cDNA microarray leads to the new fact that Belotecan, as a topoisomerase I inhibitor, is not only involved with apoptotic, cell cycle-related pathway but also involved in DNA repair.

Anticancer effect of CKD-602(belotecan, camtobell(R)) on the oral cancer cell lines

PURPOSE: CKD-602, a newly developed water-soluble campthotecin analogue, is a anticancer agent which act as a DNA topoisomerase I inhibitor. CKD-602 is known as more potent and tolerable agent. The main purposes of this study were to measure the cytotoxic effect of CKD-602 on the oral cancer cell lines and to evaluate the apoptotic aspect of dead cells. MATERIALS AND METHODS: To determine the cytotoxic effect of CKD-602 on the oral cancer cell lines in comparison with various cell lines, such as lung cancer and colon cancer cell lines, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was performed. And apoptosis was analyzed using fluorescence-activated cell sorting(FACS) system. RESULTS: CKD-602 decreased the viability of malignant cells in a dose dependent manner and in a time dependent manner. CKD-602 showed excellent cytotoxicity to the oral cancer cell lines. Also, apoptotic portion was increased in a dose dependent manner. CONCLUSION: These findings indicated that CKD-602 induced apoptotic cell death in the various cell lines including oral cancer cell lines. From the results, it was suggested that CKD-602 would be a potential therapeutic agent for the oral cancer. More successive researches on the anticancer effect of CKD-602 should be performed.

Pneumothorax as the First Clinical Manifestation of Systemic Sclerosis : A Case Report of Multiple Cystic Lung Lesions in Systemic Sclerosis / 결핵

No abstract available.

Detection of Anti-Topoisomerase I Autoantibody in Patients with Silicosis

Objectives: The aim of this study was to detect anti-topoisomerase l (anti-topo I) autoantibodies, which are known to be limited in systemic sclerosis patients, in silicosis patients with no clinical symptoms of autoimmune disease. Methods: Serum anti-topo I autoantibodies were detected using ELISA. Differences in clinical parameters between patients with and without anti-topo I autoantibodies were analyzed. Results: Seven of 69 patients had anti-topo I autoantibodies. These 7 patients showed elevated PaCO2 values (P=0.0212), and inverse correlations between serum soluble Fas levels and PaCO2 values were found. Conclusion: Anti-topo I autoantibodies were detected in 10.1% of silicosis patients without any clinical symptoms of autoimmune disease. The findings here suggest that the genesis of anti-topo l autoantibodies might be related to pulmonary involvement or lung fibrosis associated with progression of silicosis.

Detection of anti-topoisomerase I autoantibody in patients with silicosis

<p><b>OBJECTIVES</b>The aim of this study was to detect anti-topoisomerase I (anti-topo I) autoantibodies, which are known to be limited in systemic sclerosis patients, in silicosis patients with no clinical symptoms of autoimmune disease.</p><p><b>METHODS</b>Serum anti-topo I autoantibodies were detected using ELISA. Differences in clinical parameters between patients with and without anti-topo I autoantibodies were analyzed.</p><p><b>RESULTS</b>Seven of 69 patients had anti-topo I autoantibodies. These 7 patients showed elevated PaCO(2) values (P=0.0212), and inverse correlations between serum soluble Fas levels and PaCO(2) values were found.</p><p><b>CONCLUSION</b>Anti-topo I autoantibodies were detected in 10.1% of silicosis patients without any clinical symptoms of autoimmune disease. The findings here suggest that the genesis of anti-topo I autoantibodies might be related to pulmonary involvement or lung fibrosis associated with progression of silicosis.</p>

Clinical Significance of Anti-topoisomerase I Antibodies Detected by Enzyme Linked Immunosorbent Assay / 대한임상병리학회지

BACKGROUND: Anti-topoisomerase I antibodies (anti-topo-I) have been known to be a specific serologic marker for systemic sclerosis (SSc). However, anti-topo-I have also been detected fre-quently in the sera of patients with diagnosis other than SSc since the enzyme linked immunosor-bent assay (ELISA) has been used widely. In order to clarify the clinical significance of anti-topo-I on ELISA, we analyzed the clinical features of the patients positive for anti-topo-I. METHODS: Anti-topo-I and other antinuclear antibodies (ANA) were investigated by conventional ELISA methods. The clinical characteristics were analyzed in 38 patients positive for anti-topo-I and 28 patients with SLE but negative for anti-topo-I. RESULTS: Of 38 patients positive for anti-topo-I, 15 were SLE and eight SSc. The mean level of anti-topo-I in the patients with SSc was higher than that in the patients with SLE (P=0.015). Of 15 anti-topo-I positive patients with SLE, 14 had one or more other ANAs in their sera whereas only one of eight anti-topo-I positive patients with SSc did (P=0.000). There was no significant difference in clinical characteristics between anti-topo-I positive and negative patients with SLE. The preva-lences of restrictive lung disease in both groups with SLE were significantly lower than that in the anti-topo-I positive patients with SSc (P=0.008). CONCLUSIONS: Anti-topo-I is not exclusively specific for SSc and present in a considerable subset of SLE. As well as the level of anti-topo-I, the coexistence of other ANAs is helpful to discriminate SLE from SSc. The Anti-topo-I detected by ELISA does not seem to be a risk factor for restrictive lung disease in the patients with SLE, unlike those with SSc.

Topotecan-based combination chemotherapy in patients with transformed chronic myelogenous leukemia and advanced myelodysplastic syndrome

BACKGROUND: Patients with transformed chronic myelogenous leukemia(CML) and advanced myelodysplastic syndrome(MDS) have poor prognosis. The aim of this study is to evaluate the feasibility of second chronic phase induction in accelerated phase(CML-AP) or blastic crisis of CML(CML-BC) and remission induction in advanced MDS by combining topoisomerase I inhibitor (topotecan) with topoisomerase II inhibitor(mitoxantrone). METHODS: Twenty-four evaluable patients were entered on this study with a median age of 34 years. Eighteen patients with transformed CML(7 CML-AP, 11 CML-BC) and 6 patients with advanced MDS were treated. Topotecan was administered as 1.5 mg/m2/day by continuous infusion over 24 hours daily for 5 days every 4 to 8 weeks until remission. To enhance the tumoricidal effects, mitoxantrone(12 mg/m2/day, Days 1-3) was added. RESULTS: Eight patients(33+ACU-) achieved a complete remission(CR). Four of 7 patients with CML-AP(57+ACU-), 2 of 4 patients with CML-lymphoid blastic crisis (-LBC)(50+ACU-) and 2 of 6 patients with advanced MDS(33+ACU-) had CR lasting more than 45 days(45 to 400 days). There was no CR in the patients with CML-myeloid blastic crisis(-MBC). The dose level of 1.5 mg/m2/day(7.5 mg/m2/course) of topotecan was well tolerated in all patients. Mucositis occurred in 69+ACU- of patients (severe in 5+ACU-) and diarrhea in 67+ACU-(severe in 8+ACU-). In addition, there were no new or unexpected toxicities in the patients who were treated at this dose(7.5 mg/m2/course). In patients who recovered their neutrophil count, the absolute neutrophil count(ANC) remained below 500/microL for a period of 13 to 58 days(median 21 days) and the time to ANC recovery was associated with pretreatment severity of bone marrow fibrosis(mainly CML patients). Likewise, in the patients who recovered unsupported platelets, the platelets remained below 20,000/microL for a period of 0 to 37 days (median 19 days). CONCLUSION: The combination of topotecan-mitoxantrone has shown modest activity in CML-AP, CML-LBC and advanced MDS with acceptable toxicities.

Expression of Multidrug Resistant Genes in Bone Marrow Mononuclear Cells of Patients with Myeloid Leukemia / 대한암학회지

PURPOSE: Multidrug resistance mediated by several drug resistant genes impedes the successful outcome of anti-cancer chemotherapy. In this study, we investigated the expressions of drug resistant genes encoding multidrug resistance (MDR1), multidrug resistance-associated protein (MRP), topoisomerase I (Topo I), topoisomerase II g (Topo II a) in narmal volunteers (n=12) in and patients with myeloid leukemia (n=34). Material and Method: We compared the levels of their transcripts in bone matrow mononuclear cells by semiquantitative RT-PCR. The amount of specific transcripts was represented as the optical density ratio of PCR product of target gene to that of B2- microglobulin (MG). Twenty patients of acute myelogenous leukemia (eight in remission state, twelve in refractory) and fourteen patients of chronic myelogenous leukemia (nine in chronic phase and five in blastic crisis) were examined. Twelve normal healthy persons were compared with leukemic patients. RESULTS: The expression levels of all resistant genes in normal volunteers were relatively high as those of AML patients. Regardless of the disease status including remission status of AML (complete remission versus refractory) and the phase of CML (chronic phase versus blastic phase), the expression levels of all resistant genes in patients with CML were significantly lower than in the patients with AML (p < 0.05). Of interest, the patients with refractary AML did not show any statistical difference in comparison with normal controls and even the patients with AML in complete remission. Among the four drug resistant genes, the optical density ratio of MDRl was significantly lower than that of any other genes (p<0.05). Using HL-60 cell line, we compared the changes of various resistant gene expressions before and after differentiation induced by dimethylsulfoxide. The expressions of resistant genes declined in paralle1 with granulocytic differentiation, suggesting that the induction of cell differentiation might make leukemic cells susceptible to chemotherapeutic agents. CONCLUSION: It is impossibble to explain the mechanism of drug resistance by comparing the level of drug resistant gene expression between nonnal subjects and patients with myeloid leukemias. Therefore, we suppose that longitudinal study of drug resistant gene expression is necessary to demonstrate the development of drug resistant during chemotherapy.

Research progress on topoisomerase Ⅰ inhibitors / 中国药理学通报

Following the realization that involved in all of the key process within the cell nucleus such as DNA replication,transcription,reorganization,repair,human DNA topoisomerase Ⅰ(Topo Ⅰ) is a useful therapeutic target against tumor growth.Topo Ⅰ inhibitors represent a class of effective agents that have been extensively exploited and used for carcinomachemotherapy.Current Topo Ⅰ inhibitors being clinically used or developed are classified as campotothecins and non-camptothecins.This presentation introduces current status and trends of Topo Ⅰ inhibitors as anti-tumor agents with an emphasis on bioactive and pharmacological properties of these agents.