ABSTRACT
BACKGROUND: Intensified immunochemotherapy with rituximab, doxorubicin, cyclophosphamide, vindesine, bleomycin, and prednisone (R-ACVBP) improves outcomes in younger adults with diffuse large B-cell lymphomas (DLBCL) compared with R-CHOP. Due to vindesine unavailability, we assessed the safety and efficacy of replacing vindesine with vincristine in a modified R-ACVBP protocol (mR-ACVBP). METHODS: This is a retrospective study including all consecutive adult patients with newly diagnosed DLBCL who received first-line mR-ACVBP. Vindesine was replaced with vincristine 1.5 mg on days 1 and 5 of each cycle. Responders continued with published R-ACVBP consolidation. Patients with inadequate response on interim imaging were offered consolidative autologous stem cell transplantation. RESULTS: We identified 56 patients with DLBCL, with a median age of 41 years (range, 21-67). Thirty-seven (66%) patients had an age-adjusted International Prognostic Index of ≥ 2. Complete response was achieved in 41 (80%) patients and partial response in 6 (12%). The most common adverse events during induction were anemia (91%), febrile neutropenia (64%; grade 4 in 46%), thrombocytopenia (39%), and mucositis (21%). Peripheral neuropathy was encountered in 7 (12%) patients (grade 3; n = 1). Two deaths from septic shock were reported in patients with initial poor performance status. After a median follow-up of 17 months, the 2-year progression-free survival and overall survival rates were 86% and 87%, respectively. CONCLUSION: The replacement of vindesine with vincristine in mR-ACVBP seems feasible, with manageable adverse events and excellent 2-year progression-free survival. These data need validation in larger prospective trials.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Lymphoma, Large B-Cell, Diffuse/drug therapy , Vincristine/therapeutic use , Vindesine/therapeutic use , Adult , Aged , Antineoplastic Combined Chemotherapy Protocols/pharmacology , Bleomycin/pharmacology , Bleomycin/therapeutic use , Cyclophosphamide/pharmacology , Cyclophosphamide/therapeutic use , Doxorubicin/pharmacology , Doxorubicin/therapeutic use , Female , Humans , Male , Middle Aged , Prednisone/pharmacology , Prednisone/therapeutic use , Retrospective Studies , Vincristine/pharmacology , Vindesine/pharmacology , Young AdultABSTRACT
In addition to their major targets, clinically effective drugs may have unknown off-targets. By identifying such off-targets it may be possible to repurpose approved drugs for new indications. We are interested in the Golgi apparatus as a novel target for cancer therapy, but there is a paucity of candidate Golgi-disrupting drugs. Here, we aimed to identify Golgi-disrupting compounds from a panel of 34 approved anticancer drugs by using HBC-4 human breast cancer cells and immunofluorescence microscopy to visualize the Golgi apparatus. The screen identified five drugs having Golgi-disrupting activity. Four of them were vinca alkaloids (vinorelbine, vindesine, vincristine and vinblastine), and the fifth drug was eribulin. This is the first study to demonstrate that vinorelbine, vindesine and eribulin possess Golgi-disrupting activity. The 5 drugs are known to inhibit tubulin polymerization and to induce microtubule depolymerization. Interestingly, a microtubule-stabilizer paclitaxel did not induce Golgi-disruption, suggesting that the three-dimensionally preserved microtubules are partly responsible for maintaining the Golgi complex. Concerning eribulin, a noteworthy drug because of its high clinical efficacy against advanced breast cancer, we further confirmed its Golgi-disrupting activity in 3 different human breast cancer cell lines, BSY-1, MDA-MB-231 and MCF-7. Golgi-disruption may contribute to anticancer efficacy of eribulin. In conclusion, the present study revealed that 4 vinca alkaloids and eribulin possessed potential Golgi-disrupting activity among a panel of 34 approved anticancer drugs. Other drugs covering various molecular-targeted drugs and classical DNA-damaging drugs showed no Golgi-disrupting effect. These results suggest that tubulin polymerization-inhibitors might be promising candidate drugs with Golgi-disrupting activity.
Subject(s)
Antineoplastic Agents/pharmacology , Golgi Apparatus/drug effects , Tubulin Modulators/pharmacology , Antineoplastic Agents/adverse effects , Breast Neoplasms/drug therapy , Breast Neoplasms/metabolism , Cell Line, Tumor , Furans/pharmacology , Golgi Apparatus/metabolism , High-Throughput Screening Assays , Humans , Ketones/pharmacology , MCF-7 Cells , Microtubules/drug effects , Paclitaxel/pharmacology , Vinblastine/pharmacology , Vincristine/pharmacology , Vindesine/pharmacology , Vinorelbine/pharmacologyABSTRACT
OBJECTIVE: This study aimed to investigate the feasibility of intravoxel incoherent motion (IVIM) DWI and R2* (transverse relaxation rate) mapping to monitor the hyperacute therapeutic efficacy of desacetylvinblastine monohydrazide (DAVLBH) on an experimental hepatocellular carcinoma mouse model within 24 hours. MATERIALS AND METHODS: Forty-four mice were implanted with hepatocellular carcinoma and divided into three random groups. A treatment group and a control group underwent IVIM-DWI and R2* mapping examinations before and after a single injection of DAVLBH or saline at 1, 2, 4, and 24 hours. The pathology group was set for pathologic analysis, including H and E staining and CD31 and hypoxia-inducible factor (HIF)-1α immunohistochemical staining. RESULTS: DAVLBH caused hyperacute disruptions on the tumor capillaries in the treatment group. Water molecule diffusion (D), microcirculation perfusion (D*), and perfusion fraction (f) decreased initially but then gradually recovered to the baseline level by 24 hours after the first injection of DAVLBH. In contrast, R2* increased dramatically at 1 hour and then gradually decreased from 1 hour to 24 hours after treatment. D*, f, and D showed similar trends and were positively correlated with CD31 expression (r = 0.868, 0.721, and 0.730, respectively), but were negatively correlated with HIF-1α expression (r = -0.784, -0.737, and -0.673, respectively). R2* showed a negative correlation with CD31 expression (r = -0.823) and a positive correlation with HIF-1α expression (r = 0.791). CONCLUSION: Both IVIM-DWI and R2* mapping can adequately detect the vascular-disrupting effect of DAVLBH as early as 1 hour after injection in a mouse xenograft model. Moreover, D* and R2* are the two most sensitive hemodynamic parameters and can monitor the hyperacute changes associated with DAVLBH treatment in vivo.
Subject(s)
Carcinoma, Hepatocellular , Diffusion Magnetic Resonance Imaging , Liver Neoplasms , Vindesine , Animals , Female , Humans , Mice , Capillary Permeability/drug effects , Carcinoma, Hepatocellular/diagnostic imaging , Carcinoma, Hepatocellular/drug therapy , Diffusion Magnetic Resonance Imaging/methods , Disease Models, Animal , Feasibility Studies , Heterografts , Image Processing, Computer-Assisted , Liver Neoplasms/diagnostic imaging , Liver Neoplasms/drug therapy , Mice, Inbred BALB C , Microcirculation , Tumor Cells, Cultured , Vindesine/pharmacologyABSTRACT
Vinca alkaloids, the well-known tubulin-binding agents, are widely used for the clinical treatment of malignant tumors. However, little attention has been paid to their vascular disrupting effects, and the underlying mechanisms remain largely unknown. This study aims to investigate the vascular disrupting effect and the underlying mechanisms of vinca alkaloids. Methods: The capillary disruption assay and aortic ring assay were performed to evaluate the in vitro vascular disrupting effect of desacetylvinblastine monohydrazide (DAVLBH), a derivate of vinblastine, and the in vivo vascular disrupting effect was assessed on HepG2 xenograft model using magnetic resonance imaging, hematoxylin and eosin staining and immunohistochemistry. Tubulin polymerization, endothelial cell monolayer permeability, western blotting and immunofluorescence assays were performed to explore the underlying mechanisms of DAVLBH-mediated tumor vascular disruption. Results: DAVLBH has potent vascular disrupting activity both in vitro and in vivo. DAVLBH disrupts tumor vessels in a different manner than classical tubulin-targeting VDAs; it inhibits microtubule polymerization, promotes the internalization of vascular endothelial cadherin (VE-cadherin) and inhibits the recycling of internalized VE-cadherin to the cell membrane, thus increasing endothelial cell permeability and ultimately resulting in vascular disruption. DAVLBH-mediated promotion of VE-cadherin internalization and inhibition of internalized VE-cadherin recycling back to the cell membrane are partly dependent on inhibition of microtubule polymerization, and Src activation is involved in DAVLBH-induced VE-cadherin internalization. Conclusions: This study sheds light on the tumor vascular disrupting effect and underlying mechanisms of vinca alkaloids and provides new insight into the molecular mechanism of tubulin-targeting VDAs.
Subject(s)
Antigens, CD/metabolism , Cadherins/metabolism , Capillary Permeability/drug effects , Endothelial Cells/drug effects , Endothelium, Vascular/drug effects , Neoplasms/drug therapy , Vindesine/pharmacology , Animals , Cell Line, Tumor , Cells, Cultured , Endothelial Cells/metabolism , Endothelium, Vascular/metabolism , Hep G2 Cells , Human Umbilical Vein Endothelial Cells , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasms/metabolism , Rats , Rats, Sprague-DawleyABSTRACT
Multidrug resistance (MDR) plays a pivotal role in human chronic myelogenous leukemia (CML) chemotherapy failure. MDR is mainly associated with the overexpression of drug efflux transporters of the ATP-binding cassette (ABC) proteins. Phosphoinositide 3-kinase (PI3K)/Akt signaling cascade is involved in the MDR phenotype and is correlated with multidrug resistance 1 (MDR1)/P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1) and breast cancer resistance protein (BCRP) expression in many human malignancies. Homeobox (HOX) B4, a member of the HOX gene family, has been reported to be correlated with occurrence, development, poor prognosis and drug resistance of human leukemia. In the present study, HOXB4 expression was analyzed in K562 cell line and its MDR subline K562/ADM. Compared with K562 cells, drug-resistant K562/ADM cells demonstrated evidently higher HOXB4 expression. In addition, we firstly investigated the reversal effect of HOXB4 deletion on K562/ADM cells and the underlying mechanism. The Cell Counting kit-8 (CCK-8) and flow cytometry assays showed that knockdown of HOXB4 enhanced chemosensitivity and decreased drug efflux in K562/ADM cells. Moreover, HOXB4 knockout led to downregulation of P-gp, MRP1 and BCRP expression and PI3K/Akt signaling activity, suggesting that repression of HOXB4 might be a key point to reverse MDR of K562/ADM cells.
Subject(s)
Antineoplastic Agents/pharmacology , Cell Proliferation/drug effects , Drug Resistance, Multiple/genetics , Drug Resistance, Neoplasm/genetics , Homeodomain Proteins/genetics , Leukemia, Myeloid/drug therapy , Transcription Factors/genetics , ATP Binding Cassette Transporter, Subfamily B, Member 1/biosynthesis , ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , ATP Binding Cassette Transporter, Subfamily G, Member 2/biosynthesis , ATP Binding Cassette Transporter, Subfamily G, Member 2/genetics , Biological Transport/drug effects , Cell Line, Tumor , Cytarabine/pharmacology , Doxorubicin/pharmacology , Etoposide/pharmacology , Humans , Leukemia, Myeloid/genetics , Multidrug Resistance-Associated Proteins/biosynthesis , Multidrug Resistance-Associated Proteins/genetics , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , RNA Interference , RNA, Small Interfering/genetics , Reverse Transcriptase Polymerase Chain Reaction , Vindesine/pharmacologyABSTRACT
On account of its extreme intrinsic resistance to apoptosis and of its strong ability to become chemoresistant after a primary response to drugs, malignant melanoma (MM) is still a therapeutic challenge. We previously showed that glutathione S-transferase mu 1 (GSTM1) acts in synergy with multidrug resistance protein 1 (MRP1) to protect GSTM1-transfected human CAL1 melanoma cells from toxic effects of vincristine (VCR). Herein, we investigated the role of these proteins in the acquired resistance of CAL1 cells to vinca alkaloids (VAs). Resistant lines were established by continuous exposure (>1 year) of parental CAL1-wt cells to VCR, vindesine (VDS), or vinorelbine (VRB): CAL1R-VCR, CAL1R-VDS, CAL1R-VRB, respectively. All resistant lines displayed more than 10-fold increase in resistance to their selection VA, and specifically expressed GSTM1. Suggesting a direct interaction between this protein and VAs, each VA specifically decreased the GSTM1-mediated glutathione conjugation activity in cell lysates. Curcumin (GSTM1 inhibitor), BSO (glutathione synthesis inhibitor), and MK571 (MRP1 inhibitor) considerably reversed the acquired resistance to VCR and VDS, but not to VRB. Microarray data analysis revealed similar gene expression patterns of CAL1R-VCR and CAL1R-VDS, and a distinct one for CAL1R-VRB. These data suggest a differential involvement of GSTM1 and MRP1 in acquired resistance to VAs. A coordinated expression and activity of GSTM1 and MRP1 is required to protect CAL1 cells from VCR and VDS, while the simple expression of GSTM1 is sufficient, possibly by a direct drug/protein interaction, to confer resistance against VRB.
Subject(s)
Drug Resistance, Neoplasm/physiology , Glutathione Transferase/metabolism , Melanoma/drug therapy , Multidrug Resistance-Associated Proteins/metabolism , Vinca Alkaloids/pharmacology , Humans , Melanoma/metabolism , Tumor Cells, Cultured , Vinblastine/analogs & derivatives , Vinblastine/pharmacology , Vindesine/pharmacology , VinorelbineABSTRACT
Our previous study established the human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM. In this study, we investigate further the ability of tamoxifen (TAM) to reverse drug-resistance to chemotherapeutics using QBC939/ADM cells. Cell growth inhibition was determined by the MTT assay, while cell cycle progression, apoptosis and the intra-cellular concentration of adriamycin (ADM) were all determined by flow cytometry. P-glycoprotein (P-gp) protein and mRNA expression was determined by Western blotting and real-time PCR. Growth inhibition and apoptosis induced by ADM, mitomycin (MMC), or vindesine (VDS) were enhanced after pre-treatment with 5 or 10 µM TAM, while only VDS increased cell numbers in the G(2)/M phase. The intra-cellular concentration of ADM rose after pre-treatment with 10 µM TAM, but not 5 µM TAM. Furthermore, real-time PCR and western blot analysis revealed down-regulation of P-gp expression in QBC939/ADM cells after TAM pre-treatment. The enhanced effects of TAM on growth inhibition, apoptosis, and intra-cellular concentration and the down-regulation of P-gp expression were blocked by an anti-P-gp antibody. TAM (10 µM) may reverse the multi-drug-resistance (MDR) of QBC939/ADM and enhance the chemotherapeutic effects on cholangiocarcinoma, by competitively inhibiting over-expressed P-gp.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/pharmacology , Cholangiocarcinoma/pathology , Drug Resistance, Multiple/drug effects , Drug Resistance, Neoplasm/drug effects , Tamoxifen/pharmacology , ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line, Tumor , Cholangiocarcinoma/genetics , Down-Regulation/drug effects , Doxorubicin/pharmacology , Gene Expression Regulation, Neoplastic/drug effects , Humans , Inhibitory Concentration 50 , Intracellular Space/drug effects , Intracellular Space/metabolism , Mitomycin/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Vindesine/pharmacologyABSTRACT
In this study, we aim to establish the human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM which can be grow and passaged steadily in 1 µg/ml concentration of adriamycin in appropriate medium. The human multi-drug-resistant cholangiocarcinoma cell line QBC939/ADM was established using the method of exposure to medium with adriamycin alternated between high and low concentration with gradually increasing concentration. Furthermore, QBC939 and QBC939/ADM were both treated with adriamycin, mitomycin and vindesine, and then detected by MTT assay, respectively. Growth cycle and intra-cellular concentrations of ADM within cells of each group were determined by flow cytometry. Expression levels of P-glycoprotein were detected by Western bolt and real-time PCR. Results showed that, compared with QBC939, the inhibitive rates of adriamycin, mitomycin and vindesine to QBC939/ADM were lower. Content of ADM in the QBC939/ADM was lower. Western bolt and real time PCR showed that P-glycoprotein in the QBC939/ADM group was over expressed. Therefore, QBC939/ADM was establish and identified as the multi-drug-resistant cell line, which can grow and be passaged steadily in 1 µg/ml concentration adriamycin in appropriate medium. And the multi-drug-resistant character of QBC939/ADM was indicated to be related to the over expression of P-glycoprotein induced by chemotherapeutic drugs.
Subject(s)
Bile Duct Neoplasms/pathology , Bile Duct Neoplasms/physiopathology , Bile Ducts, Intrahepatic/pathology , Cell Line, Tumor/physiology , Cholangiocarcinoma/pathology , Cholangiocarcinoma/physiopathology , Drug Resistance, Multiple/physiology , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Antibiotics, Antineoplastic/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Cell Cycle/drug effects , Cell Line, Tumor/drug effects , Doxorubicin/pharmacology , Humans , Mitomycin/pharmacology , Vindesine/pharmacologyABSTRACT
BACKGROUND: The relapse of cancer after radiotherapy is a clinical knotty problem. Previous studies have demonstrated that the elevation of several factors is likely in some way to lead to the development of treatment tolerance, so it is necessary to further explore the problem of re-proliferated radioresistant cells to chemotherapeutic agents. In the present study, we aimed to investigate the chemosensitivity of radioresistant cells originated from the multicellular spheroids of A549 lung adenocarcinoma. METHODS: After irradiated with 25 Gy of 6 MV X-ray to A549 multicellular spheroids, whose 10th re-proliferated generations were employed as radioresistant cells, and the control groups were A549 parental cells and MCF7/VCR resistant cells. The chemo-sensitivity test was made by six kinds of chemotherapeutic drugs which were DDP, VDS, 5-Fu, HCP, MMC and ADM respectively, while verapamil (VPL) was used as the reversal agent. Then the treatment effect was evaluated by MTT assay, and the multidrug resistant gene expressions of mdr1 and MRP were measured by RT-PCR. RESULTS: Both A549 parental cells and A549 derived radioresistant cells were resistant to DDP, but sensitive to VDS, 5-Fu, HCP, MMC and ADM. The inhibitory rates of VPL to these two types of cell were 98% and 25% respectively (P < 0.001). In addition, without drugs added, the absorbance value (A value) of A549 parental cells was 2-folds higher than that of their radioresistant cells (P < 0.001). As to the MCF7/VCR cells, they were resistant to DDP and VDS, but slight sensitive to MMC, ADM, 5-Fu, and HCP with 80% of inhibitory rate to VPL. The subsequent RT-PCR demonstrated that the Mdr1/beta2-MG and MRP/beta2-MG of all A549 cells were about 0 and 0.7 respectively, and those of MCF7/VCR cells were 35 and 4.36. CONCLUSION: The chemosensitivity of A549 radioresistant cells had not changed markedly, and the decreased sensitivity to VPL could not be explained by the gene expression of mdr1 and MRP. It is possible that the changes in the cell membrane and decreased proliferate ability might be attributed to the resistance. Unlike multidrug resistance induced by chemotherapy, VPL may be not an ideal reverser to radioresistant cells. Therefore, the new biological strategy needs to be developed to treat recurring radioresistant tumor in combination with chemotherapy.
Subject(s)
Adenocarcinoma/drug therapy , Antineoplastic Agents/pharmacology , Drug Resistance, Neoplasm , Lung Neoplasms/drug therapy , Radiation Tolerance/drug effects , Spheroids, Cellular/drug effects , Spheroids, Cellular/radiation effects , ATP Binding Cassette Transporter, Subfamily B , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Adenocarcinoma/pathology , Adenocarcinoma/radiotherapy , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Breast Neoplasms/radiotherapy , Calcium Channel Blockers/pharmacology , Cell Proliferation , Cisplatin/pharmacology , Dose Fractionation, Radiation , Doxorubicin/pharmacology , Fluorouracil/pharmacology , Humans , Lung Neoplasms/pathology , Lung Neoplasms/radiotherapy , Mitomycin/pharmacology , Tumor Cells, Cultured , Verapamil/pharmacology , Vindesine/pharmacology , X-RaysABSTRACT
Pironetin, isolated from Streptomyces sp., is a potent inhibitor of microtubule assembly and the first compound identified that covalently binds to alpha-tubulin at Lys352. We examined whether pironetin is an effective agent against human small cell lung cancer H69 cells, including two cell lines resistant to the microtubule-targeted drugs vindesine (H69/VDS) and paclitaxel (H69/Txl) that interact with beta-tubulin. Pironetin was found to be effective against these resistant cells as well as their parental cells. In addition, pironetin inhibited the growth of human leukemic K562 multidrug-resistant cells (K562/ADM), which have mdr1 gene expression, as well as the parental K562 cells. In these cell lines, including the parental and resistant cells, pironetin caused complete mitotic arrest; in addition, apoptosis inductions by 30 and 100 nM pironetin were observed. In this study, the new mitotic inhibitor, pironetin, was found to be effective not only against human tumor cell lines resistant to microtubule-targeted drugs, but also multidrug-resistant cells with mdr1 gene expression. These results suggest that pironetin is a useful agent for overcoming drug resistance in cancer chemotherapy.
Subject(s)
Carcinoma, Non-Small-Cell Lung/drug therapy , Drug Resistance, Multiple , Lung Neoplasms/drug therapy , Paclitaxel/pharmacology , Pyrones/pharmacology , Vindesine/pharmacology , Apoptosis/drug effects , Carcinoma, Non-Small-Cell Lung/pathology , Cell Growth Processes/drug effects , Dose-Response Relationship, Drug , Doxorubicin/pharmacology , Drug Resistance, Neoplasm , Drug Screening Assays, Antitumor , HL-60 Cells , Humans , K562 Cells , Lung Neoplasms/pathology , Mitosis/drug effectsABSTRACT
Neuroblastoma (NB) is a solid childhood tumour that exhibits heterogeneous biological and clinical phenotypes. Multiple drug resistance marks a major complication especially in high-risk patients with advanced tumour stages and specific genetic aberrations, such as MYCN amplification and lp deletion. As an approach to further address the mechanisms of chemotherapeutic responsiveness of NB, we used a MYCN-inducible in vitro system and tested the susceptibility of NB cells to anti-tumour drugs currently included in NB treatment protocols dependent on MYCN expression. We observed cytotoxic effects using drug concentrations corresponding to blood plasma levels achieved in NB patients. The most potent drugs were microtubule inhibitors vindesin, paclitaxel and vincristin. Less effective were doxorubicine, arsenic trioxide, cisplatin, etoposide and carboplatin. Exposed to anti-tumour agents, NB cells with induced MYCN expression exhibited higher specific apoptosis than NB cells lacking MYCN expression. Anti-tumour drugs in MYCN-on cells accelerated G1-S phase transition, led to enhanced accumulation of cell populations in G2/M phase, and increased levels of apoptosis. In contrast, MYCN-off cell populations arrested in G1 and, to a smaller extent, in G2/M and exhibited delayed onset of apoptosis. In summary, apoptosis profiles and anti-proliferative potential of chemotherapeutic drugs, used at in vivo tolerable doses, are affected by MYCN overexpression and deregulated cell cycle in SH-EP(MYCN) cells.
Subject(s)
Antineoplastic Agents/pharmacology , Neuroblastoma/drug therapy , Nuclear Proteins/genetics , Oncogene Proteins/genetics , Apoptosis , Cell Cycle , Cell Survival/drug effects , DNA, Neoplasm/metabolism , Drug Resistance, Multiple , Gene Expression , Gene Expression Regulation, Neoplastic , Genes, myc , Humans , N-Myc Proto-Oncogene Protein , Neuroblastoma/genetics , Paclitaxel/pharmacology , Tumor Cells, Cultured , Vincristine/pharmacology , Vindesine/pharmacologyABSTRACT
In a retrospective study, O(6)-methylguanine-DNA-methyltransferase (MGMT) expression was analysed by immunohistochemistry using monoclonal human anti-MGMT antibody in melanoma metastases in patients receiving dacarbazine (DTIC) as single-drug therapy or as part of combination chemotherapy with DTIC-vindesine or DTIC-vindesine-cisplatin. The correlation of MGMT expression levels with clinical response to chemotherapy was investigated in 79 patients with metastatic melanoma. There was an inverse relationship between MGMT expression and clinical response to DTIC-based chemotherapy (P=0.05). Polymorphisms in the coding region of the MGMT gene were also investigated in tumours from 52 melanoma patients by PCR/SSCP and nucleotide sequence analyses. Single-nucleotide polymorphisms (SNPs) in exon 3 (L53L and L84F) and in exon 5 (I143V/K178R) were identified. There were no differences in the frequencies of these polymorphisms between these melanoma patients and patients with familial melanoma or healthy Swedish individuals. Functional analysis of variants MGMT-I143V and -I143V/K178R was performed by in vitro mutagenesis in Escherichia coli. There was no evidence that these variants decreased the MGMT DNA repair activity compared to the wild-type protein. All melanoma patients with the MGMT 53/84 polymorphism except one had tumours with high MGMT expression. There was no significant correlation between any of the MGMT polymorphisms and clinical response to chemotherapy, although an indication of a lower response rate in patients with SNPs in exon 5 was obtained. Thus, MGMT expression appears to be more related to response to chemotherapy than MGMT polymorphisms in patients with metastatic melanoma.
Subject(s)
Antineoplastic Agents, Alkylating/pharmacology , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Dacarbazine/pharmacology , Gene Expression Regulation, Neoplastic , Melanoma/drug therapy , Melanoma/genetics , O(6)-Methylguanine-DNA Methyltransferase/biosynthesis , O(6)-Methylguanine-DNA Methyltransferase/genetics , Polymorphism, Genetic , Skin Neoplasms/drug therapy , Skin Neoplasms/genetics , Adult , Aged , Aged, 80 and over , Antineoplastic Agents, Alkylating/administration & dosage , Antineoplastic Agents, Phytogenic/administration & dosage , Antineoplastic Agents, Phytogenic/pharmacology , Cisplatin/administration & dosage , Cisplatin/pharmacology , Dacarbazine/administration & dosage , Female , Humans , Male , Melanoma/pathology , Middle Aged , Skin Neoplasms/pathology , Treatment Outcome , Vindesine/administration & dosage , Vindesine/pharmacologyABSTRACT
In this study, we have evaluated the "in vitro" modulatory activity of a series of pyrazolotriazolopyrimidine derivatives (PTP-d) in sensitizing malignant melanoma cells to the chemotherapic drugs: taxol and vindesine. To that end, we have described the impact of chemotherapeutic agents on the cell cycle and on the induction of apoptosis when used alone or in combination with PTP-d. We have demonstrated that four PTP-d reduced chemotherapic drugs EC(50) doses of the G(2)/M accumulation with an average of 1.7-fold for taxol and 9.5-fold for vindesine when challenged on A375 human melanoma cell line. This sensitization activity was also confirmed by analyzing the apoptosis degree induced by the chemotherapic drugs. Interestingly, PTP-d had no effects on the response to cytotoxic agents by skin-derived human keratinocyte cells, NCTC 2544. Therefore, we have investigated the signaling pathway sustaining the sensitizing effect of PTP-d, providing functional evidence that active compounds are able to inhibit multidrug resistance-associated ATP-binding cassette drug transporter. These results suggested that PTP-d hold great promise for the treatment of multidrug resistance in cancers, leading to potential new therapies for melanoma.
Subject(s)
Antineoplastic Agents/pharmacology , Melanoma/pathology , Paclitaxel/pharmacology , Pyrimidines/pharmacology , Triazoles/pharmacology , Vindesine/pharmacology , Cell Cycle/drug effects , Drug Screening Assays, Antitumor , Drug Synergism , Humans , Keratinocytes/drug effects , Keratinocytes/pathology , Purinergic P1 Receptor Antagonists , Receptor, Adenosine A3 , Tumor Cells, CulturedABSTRACT
PURPOSE: The in vivo pharmacokinetics (PK), biodistribution and antitumor activity of a new liposomal formulation of gemcitabine (GemLip) were compared to the conventional (clinical) formulation of gemcitabine (GemConv). METHODS: Gemcitabine was entrapped in a vesicular phospholipid gel (VPG) consisting of densely packed liposomes. Redispersed VPG containing GemLip consisted of 33% liposomally entrapped and 67% free gemcitabine. The in vivo efficacies of GemLip and GemConv were compared using the subcutaneously growing human soft tissue sarcoma SXF 1301 and the orthotopically growing human bladder cancer BXF 1299T. PK and biodistribution were evaluated using radiolabeled drug and lipid in SXF 1301 tumor-bearing nude mice. RESULTS: GemLip was highly active in SXF 1301 at a gemcitabine dose of 6-9 mg/kg (days 1, 8 and 15; dose near the MTD). In the 6-mg/kg groups, complete tumor remissions were observed in seven of eight mice. Equimolar doses of GemConv resulted in only moderate tumor growth inhibition. Even at equitoxic doses (360 mg/kg given on days 1, 8 and 15, or 120 mg/kg on days 1, 5 and 8) GemConv was less active than GemLip. Furthermore, GemLip was active in the orthotopically growing BXF 1299T bladder cancer model at 6 mg/kg and prevented distant organ metastasis. In the PK study, GemLip achieved a 35-fold higher plasma AUC (1680 mg x h/ml) than GemConv (47.6 mg x h/ml). The serum half-lives were 0.15 h for free gemcitabine and 13.3 h for liposomal gemcitabine (6 mg/kg each i.v.). Moreover, gemcitabine levels in tumors were fourfold higher following injection of GemLip than following injection of GemConv. CONCLUSIONS: GemLip is a highly effective gemcitabine delivery system which results in superior gemcitabine pharmacodynamics and PK than GemConv. The enhanced in vivo efficacy might be explained by sustained release and passive tumor targeting.
Subject(s)
Antimetabolites, Antineoplastic/pharmacology , Antimetabolites, Antineoplastic/pharmacokinetics , Deoxycytidine/analogs & derivatives , Deoxycytidine/pharmacology , Deoxycytidine/pharmacokinetics , Paclitaxel/analogs & derivatives , Sarcoma/drug therapy , Taxoids , Urinary Bladder Neoplasms/drug therapy , Animals , Antimetabolites, Antineoplastic/administration & dosage , Antineoplastic Agents, Phytogenic/administration & dosage , Antineoplastic Agents, Phytogenic/pharmacokinetics , Antineoplastic Agents, Phytogenic/pharmacology , Deoxycytidine/administration & dosage , Docetaxel , Gels , Half-Life , Humans , Liposomes , Mice , Mice, Nude , Neoplasm Transplantation , Paclitaxel/administration & dosage , Paclitaxel/pharmacokinetics , Paclitaxel/pharmacology , Phospholipids , Tissue Distribution , Transplantation, Heterologous , Vindesine/administration & dosage , Vindesine/pharmacokinetics , Vindesine/pharmacology , GemcitabineABSTRACT
The acquisition of drug-resistance is a major problem for cancer patients undergoing chemotherapy. To clarify genetic alterations in cancer cells that develop drug-resistance, comparative genomic hybridization (CGH) was applied to esophageal squamous cell carcinoma cell lines (SH-1V1, SH-1V2, SH-1V4 and SH-1V8) and chemoresistance-related genes in altered chromosomal regions were evaluated. These cell lines were derived from the parental SH-1 cell line, after multiple steps of selection by an increasing exposure to vindesine. SH-1V8 cells were strongly resistant to vindesine. DNA copy number at 16p which includes the MRP (multidrug resistance related protein) gene was markedly increased in all cell lines examined. Increased DNA copy numbers were found at the regions of 5q31-32, 10q11.1-23, and 14q32-qter in SH-1V8 cells that acquired resistance to other drugs as well. Both SH-1V4 and SH-1V8 showed increased DNA copy numbers at 7q11.1-22, 16q12.1-qter, 19p13.2-13.3, 19q11-13.2 and 20q13.1-qter. The chromosomal region of 7q11.1-22 including MDR-1 (multidrug resistance-1) gene was highly amplified in SH-1V4 and SH-1V8. Amplification of the MRP region suggests the prerequisite of developing resistance to vindesine, and further amplification of MDR-1 may play a critical role in acquiring drug-resistance. Several unknown genes related to the induction of chemoresistance might be concealed in other altered chromosomal regions.
Subject(s)
Drug Resistance, Neoplasm/genetics , Esophageal Neoplasms/genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Vindesine/pharmacology , Chromosome Aberrations , Chromosomes, Human/genetics , Esophageal Neoplasms/drug therapy , Gene Amplification/drug effects , Gene Dosage , Genes, Neoplasm/genetics , Humans , In Situ Hybridization, Fluorescence , Metaphase/genetics , Nucleic Acid Hybridization , Physical Chromosome Mapping , Tumor Cells, CulturedABSTRACT
OBJECTIVE: To determine the chemosensitivity in fresh biopsy specimen of human oral and maxillofacial cancer, and the differential chemosensitivity among those drugs used popularly in clinic. METHODS: Human biopsy cancer cells were obtained from 150 oral and maxillofacial malignant tumors. The antitumor drugs tested using modified MTT assay were cisplatin (CDDP), 5-fluorouracil (5-Fu), Pinyangmycin (PYM), Paclitaxel (Taxol), Teniposide (Vm-26), Epi-adriamycin (E-ADM), Vindesin (VDS) and Methortrexatum (MTX). RESULTS: The success rate of the MTT assay was 93.33% (140 of the 150 cases). At a drug concentration of Cmax x 5, the inhibition rates of oral tumor cells were 63.76% for Vm-26, 25.93% for CDDP, 25.86% for E-ADM, 23.52% for Taxol, 22.97% for PYM, 22.08% for 5-Fu, 18.42% for VDS and 18.93% for MTX. The inhibition rate of VM26 was significantly higher than any of other seven chemotherapeutic drugs (P < 0.05). Over forty percent patients with squamous cell carcinoma showed moderate chemosensitivity to VM-26, CDDP and E-ADM, and over forty percent cases with adenoid carcinoma showed moderate chemosensitivity to Vm-26, Taxol and E-ADM. CONCLUSIONS: Most oral and maxillofacial cancers showed chemosensitivity to Vm-26, CDDP, E-ADM and Taxol. Vm-26, E-ADM and Taxol were more potent drugs than VDS, 5-Fu and MTX against oral and maxillofacial cancer cells. Chemosensitivity testing using modified MTT assay was useful in selecting antitumor drugs for patients with oral and maxillofacial cancers.
Subject(s)
Antineoplastic Agents/pharmacology , Maxillary Neoplasms/drug therapy , Mouth Neoplasms/drug therapy , Biopsy , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/pathology , Cell Division/drug effects , Cisplatin/pharmacology , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Fluorouracil/pharmacology , Humans , Maxilla/pathology , Maxillary Neoplasms/pathology , Mouth/pathology , Mouth Neoplasms/pathology , Paclitaxel/pharmacology , Teniposide/pharmacology , Tumor Cells, Cultured/drug effects , Vindesine/pharmacologyABSTRACT
BACKGROUND: Deletions and point mutations of the p16 gene are detectable in more than 50% of ovarian cancer cells. In this study, we examined the effect of p16 gene transduction on the growth of ovarian cancer cells and on the effect of anti-cancer agents. MATERIALS AND METHODS: p16-null human ovarian cancer cell lines, SKOV-3 and OVCAR-5, were used in this study. We transduced the full-length human p16 gene using recombinant adenovirus (AxCA-hp16). RESULTS: The spontaneous growth of these cells was significantly inhibited by hp16 transduction. MTT assay revealed that AxCA-hp16 infection induced chemoresistance in both cell lines. Flow cytometric analysis revealed that only hp16 -transduced SKOV-3, were arrested at the G1-phase for 3 days whereas those infected with AxCA-mock and OVCAR-5 infected with both recombinant viruses did not. Western blot analysis showed increased microtubule-associated proteins 4 (MAP4) in both cell lines. CONCLUSION: These results suggest that in SKOV-3 cells, G1-arrest induced by p16-transduction prevents paclitaxel- and vindesine-induced cell death, and in OVCAR-5 cells, the other unknown mechanisms play a role of chemoresistance.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Genes, p16 , Ovarian Neoplasms/drug therapy , Paclitaxel/pharmacology , Transduction, Genetic , Vindesine/pharmacology , Adenoviridae/genetics , Cell Division/drug effects , Cell Division/genetics , Combined Modality Therapy , Cyclin-Dependent Kinase Inhibitor p16/biosynthesis , Cyclin-Dependent Kinase Inhibitor p16/genetics , Cyclin-Dependent Kinase Inhibitor p16/physiology , Dose-Response Relationship, Drug , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Female , Genetic Vectors/genetics , Humans , Microtubule-Associated Proteins/biosynthesis , Ovarian Neoplasms/genetics , Ovarian Neoplasms/metabolism , Ovarian Neoplasms/pathology , Tumor Cells, CulturedABSTRACT
BACKGROUND: Oncoprotein 18 (op18) was first isolated as a molecule overexpressed in several malignant cells, suggesting a function of op18 in malignant processes, such as differentiation in hematologic malignancies, op18 also was found to enhance microtubule deassembly in the cells. Antimitotic agents that bind to tubulin have been used for chemotherapy to treat solid tumors, such as lung carcinoma. Vinca alkaloids, such as vindesine and vincristine, have commonly been used for chemotherapy of nonsmall cell lung carcinoma. The authors examined the role of op18 in the sensitivity of human lung carcinoma cells to antimitotic agents. METHODS: Expression of op18 mRNA was detected in all 17 lung carcinoma cell lines tested by Northern blotting. Oncoprotein 18 cDNA was transfected to SBC-3 human lung carcinoma cells, and the stable transfectants, SBC-3/op1-3, were isolated. The sensitivity of these transfectants against antimitotic agents were examined by the MTT assay in vitro. Cell cycle distribution of the transfectants on DNA histogram was analyzed by flow cytometry. RESULTS: Oncoprotein 18-transfected cells showed higher sensitivity to vindesine and vincristine, but not to taxanes. Vindesine-exposure increased the G2/M population of the cell cycle in the Mock transfectants, but not in SBC-3/op1, suggesting that the cell cycle dynamics were altered by op18 expression in SBC-3/op1. CONCLUSION: Oncoprotein 18 expression is associated with lung carcinoma cell sensitivity to vindesine and may be able to serve as a surrogate marker for the chemosensitivity to Vinca alkaloids in human lung carcinomas.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Carcinoma/pathology , Lung Neoplasms/pathology , Microtubule Proteins , Phosphoproteins/biosynthesis , Vindesine/pharmacology , Blotting, Northern , DNA, Complementary , Drug Screening Assays, Antitumor , Humans , Paclitaxel/analogs & derivatives , Paclitaxel/pharmacology , Phosphoproteins/pharmacology , RNA, Messenger/analysis , Stathmin , Transfection , Tumor Cells, Cultured , Vincristine/pharmacologyABSTRACT
Antineoplastics are excreted into sewage, because patients often poorly metabolize them after administration or they are metabolized into more biologically reactive metabolites. There is little information on their biodegradation and toxicity in aquatic environments. Therefore, the biodegradability of the vinca alkaloids, and their toxicity towards wastewater bacteria were investigated in this study. The biodegradability of vindesine, vincristine, and vinblastine was examined in the closed bottle test (CBT). Additionally, the biodegradability of vinblastine as a model compound of the vinca alkaloids was tested in the Zahn-Wellens test (ZWT). The growth inhibition test with Pseudomonas putida was conducted, and a toxicity control in the CBT and the ZWT was used. The colony-forming units were monitored in the CBT; the test results for the biodegradability after 28 days were: 30% for vincristine, 20% for vindesine, and 10% for vinblastine. Therefore, none of the test compounds met the criteria for being readily biodegradable (> or = 60%). Vinblastine was biodegraded up to 18% in the ZWT after 40 days, and therefore, not inherently. Toxicity towards wastewater bacteria was not found.
Subject(s)
Antineoplastic Agents, Phytogenic/metabolism , Sewage/microbiology , Vinblastine/metabolism , Vincristine/metabolism , Vindesine/metabolism , Antineoplastic Agents, Phytogenic/pharmacology , Bacteria/drug effects , Biodegradation, Environmental , Growth Inhibitors , Pseudomonas putida/drug effects , Vinblastine/pharmacology , Vincristine/pharmacology , Vindesine/pharmacology , Water MicrobiologyABSTRACT
We examined the effects of tranilast on tumor angiogenesis, tumor growth and metastasis in the mouse Lewis lung carcinoma and C57BL mouse system. Tranilast significantly reduced the dense capillary network induced by Lewis lung cancer cells in a mouse dorsal air sac angiogenesis model. Intraperitoneal administration of tranilast at 200 mg/kg/day reduced the tumor size of mouse Lewis lung carcinoma to about 63% of that of the control and suppressed pulmonary metastasis in a spontaneous system. Immunohistochemistry revealed that tranilast reduced the tumor vascularity and increased apoptosis of the tumor cells in vivo. Tranilast potentiated the inhibition of the tumor growth induced by cyclophosphamide, cis-diamminedichloroplatinum(II), adriamycin and vindesine in vivo. These results suggest that tranilast has antiangiogenic and antitumor effects and might have possible therapeutic applications.