ABSTRACT
<p><b>OBJECTIVE</b>To compare the effect of rh-endostatin on micrangium in tumor and myocardial tissue in nude mice.</p><p><b>METHODS</b>Nude mice were randomized into 4 groups (10 mice in each group), blank control group (without tumor burden, received NS 100 µl×d(-1) injection), drug control group (without tumor burden, received rh-endostatin 400 µg×d(-1) injection), model group (with tumor burden, received NS 100 µl×d(-1) injection) and treatment group (with tumor burden, received rh-endostatin 400 µg×d(-1) injection) for 28 days. The tumor volume and body weight of the mice were measured before and after administration. The expression of CD34, MMP-2, MMP-9, HIF-1α and VEGF in the myocardium and tumor were detected by immunohistochemistry. The vascular structure was observed by immunoenzymatic CD34 and Masson double staining.</p><p><b>RESULTS</b>The increase of tumor volume of the treatment group [(48.18 ± 37.31) mm(3)] was significantly lower than that in the model group [(113.80 ± 73.27) mm(3)). The changes of body weight was not significant different among the four groups. After treated with rh-endostatin, the expressions of MMP-9 and VEGF in tumors were significantly down-regulated, but the expressions of MMP-2 and HIF-1α in the tumor were not. The microvessel density (MVD) in the tumors of treatment group was significantly decreased compared with that of model group. The proportion of tumor vessels covered by collagen in the treatment group was increased compared with that of the model group. However, MVD and micrangium in myocardium were not changed significantly.</p><p><b>CONCLUSION</b>Rh-endostatin can decrease the expression of MMP-9, VEGF and MVD, inhibit the tumor growth and normalize tumor micrangium in tumor but not weaken the MMPs and MVD of mature micrangium in myocadium.</p>
Subject(s)
Animals , Female , Humans , Mice , Angiogenesis Inhibitors , Pharmacology , Antigens, CD34 , Metabolism , Antineoplastic Agents , Pharmacology , Cell Line, Tumor , Endostatins , Pharmacology , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Lung Neoplasms , Metabolism , Pathology , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Mice, Inbred BALB C , Mice, Nude , Microvessels , Pathology , Myocardium , Metabolism , Neoplasm Transplantation , Neovascularization, Pathologic , Pathology , Random Allocation , Recombinant Proteins , Pharmacology , Tumor Burden , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To observe and analyze the antitumor effect of endostar combined with docetaxel under different administration sequences.</p><p><b>METHODS</b>Nude mice with xenograft tumor (A549 cell line) were randomized into 3 groups, 8 mice/group: (1) Concurrent administration group (each mouse: endostar 400 µg/d, d1-d35, docetaxel 10 mg/kg, every 3 days, d1-d19); (2) Endo-first group (each mouse: endostar 400 µg/d, d1-d35, docetaxel 10 mg/kg, every 3 days, d16-d34); (3) Model group (positive control, tumor-bearing mice without treatment, each mouse: physiological saline, 100 µl/d, d1-d35, water for injection, 200 µl/d, d1-d35, every 3 days), and blank control group (negative control, normal mice without treatment, 8 mice), the administration method was the same to the model group. The volume of tumor and the weight of mouse were measured during treatment. Circulating endothelial cells (CECs) were detected by flowcytometry, and the expression of matrix metalloproteinase (MMP-2, MMP-9), the tissue inhibitor of MMP (TIMP-1, TIMP-2), the extracellular MMP inducer (EMMPRIN), CD34, α-smooth muscle actin (α-SMA) were determined by immunohistochemistry.</p><p><b>RESULTS</b>The tumor growth of concurrent administration group (39.94 mm(3)) was lower than that of the endo-first group [(99.57 ± 74.48) mm(3)] during treatment, both of them were smaller than that of the model group [(217.67 ± 95.44) mm(3), P < 0.05]. The amount of CECs in the endo-first group [(77.25 ± 24.02) cells/10(4) cells] was more than that of the concurrent administration group [(25.86 ± 11.77) cells/10(4) cells], the model group [(14.71 ± 11.07) cells/10(4) cells], and the blank control group [(12.90 ± 11.20) cells/10(4) cells, P < 0.01]. The expression of MMPs in the treatment groups was obviously downregulated. The expressions of TIMP-1 in the endo-first group and TIMP-2 in the concurrent administration group were upregulated (P < 0.05). The expression of EMMPRIN was significantly down-regulated in the concurrent administration group (P < 0.05). The MVD and α-SMA expressions of the treatment groups were less than that of the model group (P < 0.05).</p><p><b>CONCLUSION</b>In comparison with the endo-first group, the anti-tumor effect and survival quality of the concurrent administration group are better. Both of the administration groups may have "vascular normalization effect" by down-regulating MMPs expression through different points, and inhibit the cancer-induced stromal reaction, restraining the cancer progress to a certain extent. The changes of CECs should be a dynamic process with an initial rise in the early-stage suggesting the decrease of vascular bed and subsequent decline ascribed to apoptosis of CECs and the tumor-regression after combined therapy. Investigation of its dynamic changes may be helpful to know the change of tumor burden and vascular bed and predict the antitumor effect.</p>
Subject(s)
Animals , Female , Mice , Actins , Metabolism , Angiogenesis Inhibitors , Pharmacology , Antineoplastic Agents , Pharmacology , Antineoplastic Agents, Phytogenic , Pharmacology , Antineoplastic Combined Chemotherapy Protocols , Pharmacology , Basigin , Metabolism , Cell Line, Tumor , Drug Administration Schedule , Endostatins , Pharmacology , Endothelial Cells , Cell Biology , Lung Neoplasms , Metabolism , Pathology , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Mice, Inbred BALB C , Mice, Nude , Microvessels , Neoplasm Transplantation , Taxoids , Pharmacology , Tissue Inhibitor of Metalloproteinase-1 , Metabolism , Tissue Inhibitor of Metalloproteinase-2 , Metabolism , Tumor BurdenABSTRACT
<p><b>BACKGROUND</b>Although antiangiogenesis therapy plays an important role in anti-neoplastic treatment with its recognized efficacy and slight adverse effect, there is no prospective clinical trial to define ideal markers for predicting efficacy of antiangiogenic therapy. This study was undertaken to investigate the changes of activated circulating endothelial cells (aCECs) and survivin after anti-angiogenesis therapy and their significance in predicting the efficacy of the therapy.</p><p><b>METHODS</b>Patients of non-small cell lung cancer (NSCLC) treated with chemotherapy with or without Endostar were observed. The amount of activated CECs was detected by flow cytometry, and the expression of survivin mRNA was determined by real-time polymerase chain reaction (PCR).</p><p><b>RESULTS</b>After treatment, the amount of activated CECs decreased significantly in clinical benefit cases (P = 0.021 in chemotherapy alone, P = 0.001 in chemotherapy plus Endostar), increased in disease progressive cases (P = 0.015 in chemotherapy alone, but P = 0.293 in chemotherapy with Endotatar). After therapy, the expression of survivin mRNA decreased in clinical benefit cases (P = 0.001) and increased in disease progressive cases (P = 0.018). A positive correlation was found between activated CECs and survivin in the chemotherapy group pre- and post-therapy (P = 0.001 and 0.021, respectively), but only in the chemotherapy with Endostar group pre-therapy (P = 0.030) rather than post-therapy. A positive correlation was found between the decreased activated CECs after therapy and time to progression (TTP) (r = 0.322, P = 0.012); a negative correlation was found between the amount of survivin mRNA in serum post-therapy and TTP (r = -0.291, P = 0.048).</p><p><b>CONCLUSIONS</b>Activated CECs and survivin may be ideal markers forecasting efficacy and prognosis of NSCLC. The former can reflect more sensitively antiangiogenic efficacy and the latter is more sensitive to shrinkage or swelling of tumors. Their combination can evaluate more accurately the efficacy of antiangiogenic therapy of NSCLC.</p>
Subject(s)
Female , Humans , Male , Angiogenesis Inhibitors , Therapeutic Uses , Carcinoma, Non-Small-Cell Lung , Drug Therapy , Genetics , Metabolism , Endostatins , Therapeutic Uses , Endothelial Cells , Metabolism , Flow Cytometry , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Inhibitor of Apoptosis Proteins , Lung Neoplasms , Drug Therapy , Genetics , Metabolism , Microtubule-Associated Proteins , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To assess the effect of endostatin on growth and neoplastic angiogenesis in transplanted human lung adenocarcinoma Calu-6 tumor in nude mice.</p><p><b>METHODS</b>To treat Calu-6 tumor-bearing mice with endostatin at different doses, and to record the changes of the tumor size. The expressions of survivin, VEGF, COX-2 and MVD in tumor tissue were examined by immunohistochemistry staining, circulating endothelial cells (CECs) by flow cytometry and mRNA of CD146 and CD105 by RT-PCR and real-time PCR.</p><p><b>RESULTS</b>After endostatin treatment, the tumor size was conspicuously shrunk, and the expressions of survivin, COX-2 and VEGF protein and MVD in tumor tissue decreased concomitantly with the significant difference between each of trial groups and control group (all P < 0.05). Both CECs and mRNA of CD146 and CD105 diminished remarkably. A positive correlation between both exhibition and change of amount of activated CECs and survivin, VEGF expression and MVD count in tumor tissue was found.</p><p><b>CONCLUSION</b>Endostatin can decrease the expression of survivin, COX-2, VEGF and MVD, and to inhibit the growth of transplanted tumor. Activated CECs may probably serve as an ideal marker to predict the efficacy and prognosis of anti-angiogenesis therapy.</p>
Subject(s)
Animals , Female , Humans , Mice , Adenocarcinoma , Pathology , Angiogenesis Inhibitors , Pharmacology , Antigens, CD , Metabolism , Antineoplastic Agents , Pharmacology , CD146 Antigen , Metabolism , Cell Line, Tumor , Cyclooxygenase 2 , Metabolism , Dose-Response Relationship, Drug , Endoglin , Endostatins , Pharmacology , Endothelial Cells , Pathology , Inhibitor of Apoptosis Proteins , Lung Neoplasms , Metabolism , Pathology , Mice, Inbred BALB C , Mice, Nude , Microtubule-Associated Proteins , Metabolism , Microvessels , Pathology , Neoplasm Transplantation , RNA, Messenger , Metabolism , Random Allocation , Receptors, Cell Surface , Metabolism , Tumor Burden , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes and clinical value of circulating endothelial cells (CEC) in the peripheral blood of advanced NSCLC patient.</p><p><b>METHODS</b>Sixty-seven advanced NSCLC patients were randomly divided into either the treatment group with NP plus endostatin or control group with NP alone. Level of CEC and cytokeratin (CK) in the peripheral blood were measured by flow cytometry.</p><p><b>RESULTS</b>The response rate and benefit rate was 44.4%, 80.0% in the treatment group, and 27.3%, 50.0% in the control group, respectively (P = 0.176 and P = 0.012). Time to tumor progression (TTP) was 146.7 days in the treatment group and 91.1 days in the control group (P = 0.061). However, when the cut-off of TTP was defined as > 170 days, there was a significant difference between two groups (cut-off = 170, P = 0.034; cut-off = 180, P = 0.009). The number of CEC decreased by 0.29 +/- 0.47 in the treatment group and by 0.01 +/- 0.43 in the control group (P = 0.033). The correlation between CEC and CK was found to be positive either before (r = 0.381, P = 0.013) or after the treatment (r = 0.450, P = 0.004).</p><p><b>CONCLUSION</b>Chemotherapy combined with endostatin is superior to chemotherapy alone in the treatment of NSCLC. CEC, as a biomarker, may be useful in predicting the efficacy of the combined treatment.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Carcinoma, Non-Small-Cell Lung , Blood , Drug Therapy , Pathology , Cell Count , Cisplatin , Endostatins , Endothelial Cells , Pathology , Endothelium, Vascular , Pathology , Flow Cytometry , Follow-Up Studies , Keratins , Blood , Lung Neoplasms , Blood , Drug Therapy , Pathology , Neoplastic Cells, Circulating , Pathology , Remission Induction , Treatment Outcome , VinblastineABSTRACT
<p><b>OBJECTIVE</b>In order to explore the correlation between the centrosome aberration and oncogenesis of the breast carcinoma, the expression of alpha-tubulin and gamma-tubulin proteins in breast precancerous lesions, ductal carcinoma in situ (DCIS) and invasive ductal carcinomas (IDC) was investigated.</p><p><b>METHODS</b>Quantitative immunofluorescence analysis was performed for measuring centrosome proteins by FITC-labeled monoclonal anti-alpha and anti-gamma-tubulin antibodies in 90 cases with precancerous lesions, DCIS and IDC of the breast, respectively. Normal breast tissue from 30 cases were taken as control group.</p><p><b>RESULTS</b>The average of positive (FITC-labeled) cells were 3.2, 11.6, 14.8, 23.1 (alpha-tubulin) and 3.3, 10.7, 14.5, 24.5 (gamma-tubulin) in four groups, respectively. There were significant differences of alpha-tubulin or gamma-tubulin expression among those groups (P = 0.000), respectively. The highest expression quantity was in IDC group and the lowest was in normal breast tissue. Their expression was significantly associated with cellular proliferation and differentiation.</p><p><b>CONCLUSION</b>There is over-expression of the centrosome tubulin protein in the precancerous stage of the breast. The centrosome aberration may play an important role during the crucial early step of oncogenesis and it may promote the cellular cancerization or transformation into malignancy. Quantitative immuno-fluorescence analysis and immunohistochemistry can be complementary each other.</p>