ABSTRACT
Arsenic trioxide (As2O3) was introduced into the treatment of refractory or relapsed acute promyelocytic leukemia and showed a striking effectiveness in China and United States multicenter study. However, the mechanistic basis for the carcinogenic or therapeutic effects of arsenics is still poorly understood. So, this study is performed to determine whether As2O3 induces apoptosis through intrinsic caspase cascades in acute promyelocytic leukemia HL-60 cells. MATERIALS AND METHODS: HL-60 cells were treated with As2O3 to investigate apoptosis through signaling of caspase cascades and mitochondrial dysfunction. RESULTS: As2O3 (>0.5 uM) decreased the viability of HL-60 cells in a dose-dependent manner, which was revealed as apoptosis shown chromatin condensation and ladder pattern DNA fragmentation. As2O3 increased the catalytic activity of caspase family cysteine proteases including caspase-3 and -9 proteases. Consistently, PARP, an intracellular biosubstrate of caspase-3 protease, was cleaved from 116 kDa to 85 kDa fragments. It also induced the change of mitochondrial membrane potential. Morever, As2O3 resulted in the increase of Bak. CONCLUSION: These data suggest that As2O3 induces apoptosis of HL-60 cells through activation of intrinsic caspase protease with mitochondrial dysfunction.
Subject(s)
Humans , Apoptosis , Arsenic , Caspase 3 , China , Chromatin , Cysteine Proteases , DNA Fragmentation , HL-60 Cells , Leukemia, Promyelocytic, Acute , Membrane Potential, Mitochondrial , Peptide Hydrolases , Strikes, Employee , United StatesABSTRACT
Nitric oxide (NO) induces apoptotic cell death in murine RAW 264.7 macrophages. To elucidate the roles of SEK1/MKK4, a upstream kinase for both c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) and p38 kinase, on NO-induced apoptosis, we generated clones of RAW 264.7 cells which stably overexpressd kinase inactive SEK1 (RAW/SEK1-Kl) or wild type SEK1 (RAW/SEK1-WT). Treatment of kinase inactive SEK1 transfected RAW 264.7 cells (RAW/SEK1-Kl) with sodium nitroprusside (SNP), a NO generating agent, significantly decreased the cell viability up to 20% of RAW control cells which were treated with the same amount of SNP. However, RAW/SEK1-WT cells were less susceptible to NO induced apoptosis. For a while, caspase-3 like activity in NO treated RAW/SEK1-Kl cells was significantly increased with parallell to apoptotic death rate. However, caspase1 like activity was not affected by NO in any transfectants. The NO induced apoptosis in RAW/SEK1-Kl cells was significantly prevented by the addition of caspase-3 like inhibitor (N-Ac- DEVD-CHO). In addition, the phosphotransferase activity of JNK1 in NO-treated RAW/SEK1-WT is significantly increased, but not in RAW/SEK1-Kl cells. These results suggest that SEK1 may play anti-apoptotic role in RAW cells from NO-induced apoptosis.
Subject(s)
Apoptosis , Caspase 3 , Cell Death , Cell Survival , Clone Cells , Macrophages , Mortality , Nitric Oxide , Nitroprusside , Phosphotransferases , Protein KinasesABSTRACT
Gliotoxin, a fungal metabolite, is one of the epipolythiodioxopiperazine classes and has a variety of effects including imrnunomodulatory and apoptotic agents. This study is designed to evaluate the effect of zinc on gliotoxin-induced death of HL-60 cells. Here, we demonstrated that treatment of gliotoxin decreased cell viability in a dose and time-dependent manner. Gliotoxin-induced cell death was confirtned as apoptosis characterized by chromatin marginafion, fragmentation and ladder-pattern digestion of genomic DNA. Gliotoxin increased the proteolytic activities of caspase 3, 6, 8, and 9. Caspase-3 activation was further confirmed by the degradation of procaspase-3 and PARP in gliotoxin-treated HL-60 cells. Zinc compounds including ZnC12 and ZnSO4 markedly inhibited gliotoxin-induced apoptosis in HL-60 cells (from 30% to 90%). Consistent with anti- apoptotic effects, zinc also suppressed the enzymatic activities of caspase-3 and -9 proteases. In addition, cleavage of both PARP and procaspase 3 in gliotoxin-treated HL-60 cells was inhibited by the addition of zinc compounds. We further demonstrated that expression of Fas ligand by gliotoxin was suppressed by zinc compounds. These data suggest that zinc may prevent gliotoxin- induced apoptosis via inhibition of Fas ligand expression as well as suppression of caspase family cysteine proteases-3 and -9 in HL-60 cells.
Subject(s)
Humans , Apoptosis , Caspase 3 , Cell Death , Cell Survival , Chromatin , Cysteine , Digestion , DNA , Fas Ligand Protein , Gliotoxin , HL-60 Cells , Peptide Hydrolases , Zinc Compounds , ZincABSTRACT
Bacterial, fungal, and viral infections of the skin with extended skin involvement can occur during the early phase of human immunodeficiency virus infection. A significant reduction in circulating CD4+ lymphocytes in the late stage of the disease may cause tumors of the skin such as Kaposi's sarcoma. A 40-year male patient, a former sailor who had multiple sexual contact with native African women, presented with multiple tender follicular pustules and fibrotic brown patches on both his legs. these had been present for 6 months. The skin lesions were healed leaving brown pigmentation. Laboratory examinations revealed the presence of leukopenia, thromocytopenia and a reversed T4/T8 ratio. The ELISA and Western blot analysis to human immunodeficiency virus were positive. A skin biopsy from a brown patch showed early stages of scar tissue and perivascular hemosiderin deposition. We herein report a case of acquired immunodeficiency syndrome patient with atypical dark brown scarring atrophic patches on the lower legs following purulent bacterial folliculitis. This may have been an early manifestation of Kaposi's sarcoma from a preceding skin lesion.
Subject(s)
Female , Humans , Male , Acquired Immunodeficiency Syndrome , Biopsy , Blotting, Western , Cicatrix , Enzyme-Linked Immunosorbent Assay , Folliculitis , Hemosiderin , HIV , Leg , Leukopenia , Lymphocytes , Military Personnel , Pigmentation , Sarcoma, Kaposi , SkinABSTRACT
Alpha-storage pool deficiency of platelet is a very rare disorder associated with a variety of conditions, including autoimmune disease, disseminated intravascular coagulation, myeloproliferative disorders, and cardiopulmonary bypass. This bleeding disorder is characterized by a moderate thrombocytopenia and a selective abnormality in platelet alpha-granules. We experienced the first case of alpha-storage pool deficiency of platelet in a 28-year-old male with severe valvular heart disease in Republic of Korea. Two years ago, mitral valve replacement was performed in other university hospital. Since a few months ago, dyspnea was developed and progressively exacerbated. Transesophageal echocardiogram showed severe mitral valve regurgitation and aortic valve regurgitation. He had moderate thrombocytopenia. Bleeding time was prolonged. Peripheral blood smear showed hypogranular platelets with indistinct cell membrane. In bone marrow biopsy, there showed evidence of mild hyperplasia of megakaryocytes. Platelet aggregation test revealed abnormal response to collagen and epinephrine. Electron microscopy of periphral blood showed the vacuolization of alpha- granules of platelets. Following platelet transfusion, double valve replacement could be performed successfully. Six months later, platelet morphology was moderately normalized. But bleeding time and platelet aggregation test were still abnormal.
Subject(s)
Adult , Humans , Male , Aortic Valve , Autoimmune Diseases , Biopsy , Bleeding Time , Blood Platelets , Bone Marrow , Cardiopulmonary Bypass , Cell Membrane , Collagen , Disseminated Intravascular Coagulation , Dyspnea , Epinephrine , Heart Valve Diseases , Hemorrhage , Hyperplasia , Megakaryocytes , Microscopy, Electron , Mitral Valve , Mitral Valve Insufficiency , Myeloproliferative Disorders , Platelet Aggregation , Platelet Storage Pool Deficiency , Platelet Transfusion , Republic of Korea , ThrombocytopeniaABSTRACT
OBJECTIVES: The poor survival rates among patients receiving surgery alone for stages II and III non-small cell lung cancer prompted several trials of adjuvant therapy after resection. We performed a prospective phase II study in patients with stage II-IIIA non-small cell lung cancer after resection to evaluate the feasibility, activity and toxicity of the postoperative sequential MVP chemotherapy and radiotherapy. METHODS: Between February 1991 and May 1995, 60 patients with resected stage II, IIIA non-small cell lung cancer received 2 cycles of MVP combination chemotherapy (Mitomycin-C 6 mg/m2, Vinblastine 6 mg/m2, Cisplatin 60 mg/m2) within 3 weeks after surgery, followed by thoracic irradiation (5,040 cGy after complete resection and 900 cGy booster to microscopically positive resection margin at 1.8 Gy per fraction) within 3-4 weeks after chemotherapy. RESULTS: Forty nine men and 11 women with a median age of 60.5 years (range 33-81 years) were included. During the median follow-up period of 828 days (61-2,015 days), 25 patients had developed recurrence. Among the 25 failures, 3 were local relapse only and 20 were distant metastasis only and 2 had both local and distant sites of recurrence. Three-year overall survival and event-free survival were 43% and 37%, respectively. Neutropenia of grade I-II was observed only in 13 patients. Eleven patient showed grade I-II radiation pneumonitis and 32 had grade I-II radiation esophagitis. CONCLUSION: Postoperative sequential MVP chemotherapy and radiotherapy in resected stage II-IIIA non-small cell lung cancer is well-tolerated and shows interesting activity.
Subject(s)
Female , Humans , Male , Carcinoma, Non-Small-Cell Lung , Cisplatin , Disease-Free Survival , Drug Therapy , Drug Therapy, Combination , Esophagitis , Follow-Up Studies , Mitomycin , Neoplasm Metastasis , Neutropenia , Prospective Studies , Radiation Pneumonitis , Radiotherapy , Recurrence , Survival Rate , VinblastineABSTRACT
PURPOSE: Recent studies indicate that widely used chemotherapeutic agents induce apoptosis in susceptible cells. One of the effector arms in this cell death pathway is composed of cysteine proteases belonging to the caspase family. In cells, caspase-3 has been shown to play an important role as a downstream member of protease cascade, where various cell death pathways converge into the same effector pathway. JNK, a member of the mitogen-activated protein kinase pathway, is activated in response to many stressful stimuli including heat shock, UV irradiation, protein synthesis inhibitors, and inflammatory cytokines. In this study, we investigated whether JNK1 & caspase-3 play a role in the apoptosis induced by adriamycin (ADR). METHODS: U937 cells were cultured in RPMI 1640 and treated with different concentrations of ADR. Cellular DNA was extracted and analyzed by electrophoresis on a 1.5% agarose gel to detect DNA fragmentation. The activity of caspase-3 was measured by the proteolytic cleavage of the fluorogenic substrate DEVD-AMC. The activity of JNK1 was measured by in vitro immunocomplex kinase assay with 2 microgram of GST-c Jun as a substrate and quanititated using phosphoimager analyzer. RESULTS: ADR induced the apoptotic death of U937 myeloid cells in a dose-dependent manner, which was characterized by increasing ladder-pattern DNA fragmentation. Consistent with apoptotic death of U937 cells, ADR induced the catalytic activation of caspase-3 as well as JNK1 at 2.5 microgram/mL of concentrations. CONCLUSION: Adriamycin induces apoptosis of human myeloid leukemic U937 cells via activation of caspase-3 and cJun-N terminal kinase1 (JNK1)/Stress activated protein kinase (SAPK).
Subject(s)
Humans , Apoptosis , Arm , Caspase 3 , Cell Death , Cysteine Proteases , Cytokines , DNA , DNA Fragmentation , Doxorubicin , Electrophoresis , Fluorescent Dyes , Hot Temperature , Myeloid Cells , Phosphotransferases , Protein Kinases , Protein Synthesis Inhibitors , Sepharose , Shock , U937 CellsABSTRACT
BACKGROUND: Although small cell lung cancer is a chemosensitive disease, it grows rapidly and relapses frequently. Even with optimum treatment, only small portion of patients have experienced long-term survival. The objective of this study was to describe the clinical characteristics and therapeutic features, and to analyze the prognosis in small cell lung cancer. METHODS: We analyzed retrospectively 151 evaluable patients with histologically confirmed small cell lung cancer from August 1989 to June 1995 at our institution. Of 151 patients, 3 had surgery and chemotherapy, 59 had chemotherapy and chest irradiation, and 89 had chemotherapy only. RESULTS: Most patients(82.1%) were men, and the median age was 62 years. Of all patients, 49% had performance status of 0,1 and 59.6% had limited disease. The overall response rate was 67.8% : complete response 23.8%, partial response 44.4%. Complete responses were documented in all of three patients who had surgery and chemotherapy, 49.2% of those who had chemotherapy and radiotherapy, and 4.5% of those who had chemotherapy only. The median follow-up duration was 309 days. The median progression-free survival and overall survival were 256 days and 354 days, respectively: patients who had surgery and chemotherapy were 1631 days and 1631 days, those who had chemotherapy and radiotherapy were 344 days and 450 days, and those who had chemotherapy only were 186 days and 278 days, respectively; complete responders were 580 days and 710 days, partial responders were 231 days and 364 days, non-responders were 132 days and 151 days, respectively. Of 151 patients, 11.3% survived more than two years(long-term survival). Most long-term survivors had limited disease(82.4%) and good performance(76.5%). Long-term survival occurred in two patients of those who had surgery and chemotherapy, 16.9% of those who had chemotherapy and radiotherapy, and 5.6% of those who had chemotherapy only. Most long-term survivors(70.6%) had complete response. Twenty of 36 complete responders and 8 of 17 long-term survivors had disease relapses or progressions. Patients with limited disease, those with good performance, and those with normal alkaline phosphatase had a significantly higher complete response rate and longer progression-free survival and overall survival than their counterparts. Of pretreatment characteristics, stage and performance status were correlated complete response and survival, independently. complete response outcome was significant independent variable for survival. CONCLUSION: The disappointing results in this disease support the need for both new treatment strategies to improve complete response rate and to decrease relapse rate and large-scaled prospective studies to know natural history of long-term survivors.
Subject(s)
Humans , Male , Alkaline Phosphatase , Disease-Free Survival , Drug Therapy , Follow-Up Studies , Natural History , Prognosis , Radiotherapy , Recurrence , Retrospective Studies , Small Cell Lung Carcinoma , Survivors , ThoraxABSTRACT
We experienced a 61-year old man with Pneumocystis carinii pneumonia who had been diagnosed as having relapsed acute myelogenous leukemia(AML). He developed severe dyspnea in the nadir state after reinduction chemotherapy. His chest X-ray showed bilateral interstitial pneumonia in both lung fields. We started ventilator therapy and obtained sputum through the endotracheal tube. Typical P. carinii cysts were found in the sputum by Giemsa stain. No other organisms were found in thelavage sediments. From clinical observation and the presence of typical P. carinii cysts, the patient was diagnosed as having P. carinii pneumonia and was treated with sulfamethoxazole/trimethoprim and glucocorticoid. This was the first reported case of P. carinii pneumonia in an AML patient undergoing chemotherapy in Korea.
Subject(s)
Humans , Middle Aged , Azure Stains , Drug Therapy , Dyspnea , Korea , Leukemia , Leukemia, Myeloid, Acute , Lung , Lung Diseases, Interstitial , Pneumocystis carinii , Pneumocystis , Pneumonia , Pneumonia, Pneumocystis , Sputum , Thorax , Ventilators, MechanicalABSTRACT
OBJECTIVES: Recently high dose chemotherapy with autologous peripheral blood stem cell transplantation (APBSCT) has been investigated with the hope of maximizing tumor response and increasing survival. The purpose of this study is to evaluate the effect, feasibility, and toxicity of high-dose cyclophosphamide, thiotepa, and carboplatin (CTCb) with APBSCT in patients with metastatic or high risk primary breast cancer. METHODS: Four cases of high-risk primary breast cancer (with more than 10 involved axillary nodes) and three cases of metastatic disease in complete or partial response were enrolled. Peripheral blood stem cells were mobilized by G-CSF plus chemotherapy, and median number of collected mononuclear cells was 5.44 X 108/kg(range, 1.95-7.08 X 108/kg). High-dose chemotherapy of cyclophosphamide (1,500mg/m2/day), thiotepa (125mg/m2/day) and carboplatin (200mg/m2/day) was administered for 4 days and peripheral blood stem cells were reinfused to the patients 72 hours after the completion of chemotherapy. RESULTS: The median days of recovery for neutrophil (over 500/mm3) and for platelet (over 50,000/mm3) were 10 (range, 8 to 33) and 30 (range, 10 to 40). One patient suffered from seizure attack and grade 3 hepatotoxicity during high dose chemotherapy, There were no treatment-related death. Four patients with high-risk primary breast cancer remained disease-free at 2, 8, 12 and 19 months post-transplant. In one patient with bone metastasis, complete response was induced following APBSCT. All three patients with metastatic disease remained progression-free at 8, 18 and 19 months post-transplant. CONCLUSION: High-dose chemotherapy and autologous peripheral blood stem cell transplantation was feasible and would be a potentially effective treatment modality in high risk and metastatic breast cancer.
Subject(s)
Humans , Blood Platelets , Breast Neoplasms , Breast , Carboplatin , Cyclophosphamide , Drug Therapy , Granulocyte Colony-Stimulating Factor , Hope , Neoplasm Metastasis , Neutrophils , Peripheral Blood Stem Cell Transplantation , Seizures , Stem Cells , ThiotepaABSTRACT
BACKGROUND: To study the prognosis of patients with lung cancer, many investigators have reported the methods to detect cell proliferation in tissues including PCNA, thymidine autoradiography, flow cytometry and Ki-67. PCNA, also known as cyclin, is a cell related nuclear protein with 36KD intranuclear polypeptide that is maximally elevated in S phase of proliferating cells. In this study, PCNA was identified by paraffin-embedding tissue using immunohistochemistry which has an advantage of simplicity and maintenance of tissue architecture. The variation of PCNA expression is known to be related with proliferating fraction, histologic type, anatomic(TNM) stage, degree of cell differentiation, S-phase fraction and survival rate. We analyzed the correlation between PCNA expression and S-phase fraction, survival. METHODS: To investigate expression of PCNA in primary lung cancer, we used immunohistochemical stain to paraffin-embedded sections of 57 resected primary non-small cell lung cancer specimen and the results were analyzed according to the cell type, cell differentiation, TNM stage, S-phase fraction and survival. RESULTS: PCNA expression was dMded into five group according to degree of staging(-, +, ++, +++,++++). Squamous cell type showed high positivity than in adenocarcinoma. Nonsignificant difference related to TNM stage was noticed. Nonsignificant difference related to degree of cell differentiation was noticed. S-phase fraction was increased wit advance of PCNA positivity, but t could not reach the statistic significance. The 2 year survival rate and median survival time were -50% 13 months, +75% 41.3 months, ++73% 33.6 months, +++67% 29.0 months, ++++25% 9 months with statistic significance (P<0.05, Kaplan-Meier, generalized Wilcox). CONCLUSION: From this study. PCNA expression was high positive n squamous cell cancer. And, there was no relationship between PCNA positivity and TNM stage, cellular differentiation or S-phase fraction. But, the patients with high positive PCNA staining showed poor survival rate than the patients with lower positive PCNA. It was concluded that PCNA immunostaining is a simple and useful method for survival prediction in paraffin embedded tissue of non-small cell lung cancer.
Subject(s)
Humans , Adenocarcinoma , Autoradiography , Carcinoma, Non-Small-Cell Lung , Cell Differentiation , Cell Proliferation , Cyclins , Flow Cytometry , Immunohistochemistry , Lung Neoplasms , Neoplasms, Squamous Cell , Nuclear Proteins , Paraffin , Prognosis , Proliferating Cell Nuclear Antigen , Research Personnel , S Phase , Survival Rate , ThymidineABSTRACT
BACKGROUND: Flow cytometric study has been used to measure the DNA content of solid tumors for the last decade. DNA ploidy is an important property commonly measured by flow cytometry. The possibility to study archival paraffin-embedded tumors has hastened an appreciation of prognostic utility of this method. The aim of this study is to look for biologic prognostic indicator for survival time of patients with small cell carcinoma of lung in addition to the well known clinical prognostic factors. METHOD: DNA ploidy was measured by flow cytometric method using tumor cells isolated from paraffin embedded tissue. To evaluate the prognostic significance, DNA ploidy of small cell lung cancer was analysed in 42 patients who died after receiving anticancer chemotherapy. RESULTS: 1) Mean survival time of all patients was 190(+/-156) days. Survival time was shortened, when TNM stage and PS scale were advanced. 2) 62% of all patients was DNA aneuploidy. DNA ploidy had nothing to do with advance of TNM stage and PS scale. 3) Mean survival time of aneuploid tumor was significantly shorter(138+/-90 days) than that of diploid tumors(272 +/- 197 days).(p <0.001) 4) To exclude the influence of clinical prognostic factors such as TNM stage and PS scale, the analysis was restricted to subgroups of identical stage. We were able to find the same tendency. CONCLUSION: DNA ploidy is an independent prognostic factor in small cell lung cancer.
Subject(s)
Humans , Aneuploidy , Carcinoma, Small Cell , Diploidy , DNA , Drug Therapy , Flow Cytometry , Lung , Paraffin , Ploidies , Small Cell Lung Carcinoma , Survival RateABSTRACT
BACKGROUND: DNA content analysis of human solid tumor is now widely performed by flow cytometric study. One of the most interesting and potentially observation in this field is that proliferative activity(S-Phase fraction of cell cycle) may profoundly affect the prognosis. METHOD: S-Phase fraction(SPF) have been measured by flow cytometric method using tumor cells isolated from paraffin embedded tissue. To evaluate the prognostic significance, SPF of small lung cancer cell was assessed in 42 patients who died after receiving anticancer chemotherapy. RESULTS: 1) Mean survival time of patients with small cell lung cancer was 190(± 156) days, Survival time were shortened, when TNM stage and PS scale were advanced. 2) Mean value of SPF of patients with small cell lung cancer was 27.4(±8.5)%. SPF had nothing to do with advance of TNM stage and PS scale. 3) In each identical TNM stage, there were not statistic significance between SPF and survival times. 4) There was a tendency like that higher SPF, better chemotherapeutic CONCLUSION: We could not find statistic significance between SPF and survival times, but SPF was a good predictive factor for chemotherapeutic response.