- Email: [email protected]
Increasing serum levels of basic fibroblast growth factor in non small cell lung cancer indicates progressive disease
Biology greater than in ELF from smokers and nonsmokers. Furthermore Giutathione has been shown to modulate the cytotoxicity of a variety of chemotherapeutic agents. We examined the relationship between Glutathione levels and chemotherapy. Material and Methods: Bronchoalveolar lavage (BAL) was perfomed on 15 patients with locally advanced NSCLC before and after treatment with 2 cycles neoadjuvant chemotherapy containing mitomycin, vindesin and ifosfamid. The volume of the ELF and the concentration of glutathione and oxidized glutathione were determined. Concentrations of total glutathione in BAL and plasma were quantified spectrophotometrically, the volume of ELF recovered by BAL was determined using the urea method. In addition we looked for iL6 and TNF in BAL and plasma. Patients underwent respiratory function tests before and after therapy. Chemotoxicity, response rate and median survival rate were evaluated. Results: The percentage return of instilled saline was pretherapy 44% ± 18, posttherapy 48% + 10. The mean ELF volume recovered was similar before and after chemotherapy (0.37 ml i 0.22 versus 0.39 ml + 0.16). Total glutathione concentration in ELF changed little after chemotherapy (540.4 nmol/ml ± 315.8 versus 694.0 nmol/ml ± 595.6). Individual variability was remarkable. Discussion: Previous studies have shown that GSH can function to reduce the cellular toxicity of various chemotherapy agents. So far our results don't show effects on glutathione levels after chemotherapy. We can not conclude that antioxidant therapy may influence pulmonary toxicity of chemotherapeutic agents. Smoking induced changes in antioxidant system seem to be greater then chemotherapeutic effects. However, very little is known about the respiratory tract antioxidant defense of patients with lung cancer.
l-6--~ MMP and TIMP expression in orthotopic lung cancer models for preclinical screening of novel MMP inhibitors J. Liu, M. Johnston, J. Fata, R. Khokha, N. Liu, M. Tsao. Ontario
Cancer Institute and the Departments of Surgery and Pathology, University of Toronto, Toronto, Canada We developed orthotopic lung cancer models in nude rats by endobronchial implantation of the human non-small cell lung cancer (NSCLC) cell lines NCI-H460, NCI-H125, A549. A discreet right caudal lobe tumor develops within 3 to 10 weeks, depending on cell type. Expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMPs) was studied. Representative samples of each cell line and its respective orthotopic lung tumor were analyzed with northern blot and zymography, eDNA for MMP-1, -2, -9, -11, and MT1-MMP (MMP-14), and inhibitors TIMP-1,-2,-3, -4 were used to detect mRNA expression. All orthotopic lung tumors consistently express MMP-2, MT1-MMP and TIMP-2. The parent cell lines demonstrate significantly lower (H125, A549) or no (H460) MMP-2 expression. When fragments from an H460 orthotopic lung tumor are endobronchially implanted in a nude rat a primary lung tumor develops, followed by regional metastasis to mediastinal lymph nodes and systemic metastases to bone, brain, left lung and kidney. This pattern closely simulates the clinical behavior of NSCLC. In this model both the primary lung tumor and metastatic tumor in bone and left lung express MMP-2, MT1MMP, and TIMP-2, with the metastatic lesions tending to have a higher level of expression. Zymographic analysis revealed high levels of proMMP-2 expression in beth primary and metastatic tumors. MMP-2, MT1-MMP, and TIMP-2 may play a critical role in tumor invasion and metastasis in these models. Prinomastat (AG3340), a novel synthetic MMP inhibitor mainly targeting gelatinases (MMP-2 and -9), and MT1-MMP, was tested in our H460 metastatic orthotopic lung cancer model. Prinomastat (AG3340) (100 mg/kg po bid) in combination with carboplatin (10 mg/kg ip weekly) significantly prolonged survival compared to either drug alone. Toxicity was minimal and tolerance much better than cytotoxic agents tested in this model. These orthotopic lung cancer models may be useful for studying complex interaction of MMPs and TIMPs in vivo. Particularly, the H460 orthotopic model that metastasizes widely may be an effective preclinical screening tool to assess the efficacy of novel
203
MMP inhibitors specific for the MMPs and TIMP expressed in these tumors.
~ - - ~ Increasing serum levels of basic fibroblast growth factor in non small cell lung cancer indicates progressive disease D. Brattstr6m 1, M. Bergqvist 1, P. Hesselius 1, A. Larsson 2, G. Wagenius 3, O. Brodin 3. 1Department of Oncology, Akademiska
Hospital, Uppsala Universi~ 751 85 Uppsala; 2Department of Clinical Chemistry, Akademiska Hospital, Uppsala University, 751 85 Uppsala; 3Department of Oncology, Uppsala Akademiska Hospital, Uppsala, Sweden Basic fibroblast growth factor (bFGF) is a mitogen with many properties; one of them is to stimulate angiogenesis in a very potent manner. In vitro and in vivo studies have implicated that bFGF and VEGF are closely correlated and act synergistically in inducing angiogenesis, yet maybe in different temporal fashion. Our earlier study on S-bFGF indicates that S-bFGF have prognostic information of great importance for operable non-small cell lung cancer (NSCLC). In the present study we focused on the impact of alterations of S-bFGF during treatment in patients with inoperable NSCLC. We analysed 486 serum samples from 72 patients with inoperable NSCLC for bFGF values. For the detection of S-bFGF we used a commercially available immunosorbent assay (R&D Systems Inc., Minneapolis, MN, USA). All patients gave informed consent prior collection and a minimum of three samples were collected during the course of treatment. Thirty-eight patients were staged as Ilia, 24 patients as IIIb and 10 patients as stage IV. Mean value of S-bFGF in all samples (no 486) was 9.6 pg/ml (confidence interval (CI) 8.6-10.6). Mean values of S-bFGF in samples stratified into stage were in Ilia 8.5 pg/ml (CI 5.8-11.1), stage IIIb 11.8 pg/ml (CI 6.3-17.4) and for stage IV 12.0 pg/ml (CI 5.3-18.6). Regression analysis using a time-index statistical model demonstrated a significant correlation between increasing values of S-bFGF and date of death (p < 0.005). Regression analysis also demonstrated a significant correlation between platelets and bFGF, e.g. with each rise of 1 x 109 in platelet count, S-bFGF rises 0.021 pg/ml (p < 0.0001). When analysing location of metastases and S-bFGF values, no differences could be demonstrated between brain and skeleton compared to no sign of metastases. But, when multiple metastases and/or liver metastases were present there was a difference between S-bFGF (mean 14.7 pg/ml CI 10.4-18.9) compared to no sign of metastases (mean 9.2 pg/ml CI 8.2-10.3).
[-6--9-~ HER-2 status and response to chemotherapy in non small lung cancer J.F. Morere, M. Kambouchner, M.C. Pailler, S. Piperno-Neumann, E. Goguel, A. Martin, J.L. Braau. Avicenne Hospital, Bobigny, France Background: HER-2/neu gene overexpression may be important in cancer progression. Numerous studies have investigated the relationship between HER2 status and prognosis or response to therapies mostly in breast cancer. Methods: In this retrospective study we have evaluated the correlation of HER-2 status and the response to chemotherapy in 50 patients with advanced adenocarcinoma or large cell lung cancer. Bronchial biopsies were obtained by fibroscopy or surgery. Paraffin embedded tumor tissue were stained with an antibody to a site of the internal domain of the HER-2 receptor (CB11). Results: Overall response rate to standard chemotherapy in the whole population was 26%. Fourty three biopsies could be stained. Weak positive staining was observed in 16 out of 43 (37%). No statistically significant difference was observed in response to chemotherapy according to HER-2 status. Conclusion: In our population of patients, HER-2 staining with CB11 does not correlate with response to chemotherapy. A prospective study combining immunohistochemistry, and fluorescence in situhybridisation is ongoing.
l-6--~ MMP and TIMP expression in orthotopic lung cancer models for preclinical screening of novel MMP inhibitors J. Liu, M. Johnston, J. Fata, R. Khokha, N. Liu, M. Tsao. Ontario
Cancer Institute and the Departments of Surgery and Pathology, University of Toronto, Toronto, Canada We developed orthotopic lung cancer models in nude rats by endobronchial implantation of the human non-small cell lung cancer (NSCLC) cell lines NCI-H460, NCI-H125, A549. A discreet right caudal lobe tumor develops within 3 to 10 weeks, depending on cell type. Expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMPs) was studied. Representative samples of each cell line and its respective orthotopic lung tumor were analyzed with northern blot and zymography, eDNA for MMP-1, -2, -9, -11, and MT1-MMP (MMP-14), and inhibitors TIMP-1,-2,-3, -4 were used to detect mRNA expression. All orthotopic lung tumors consistently express MMP-2, MT1-MMP and TIMP-2. The parent cell lines demonstrate significantly lower (H125, A549) or no (H460) MMP-2 expression. When fragments from an H460 orthotopic lung tumor are endobronchially implanted in a nude rat a primary lung tumor develops, followed by regional metastasis to mediastinal lymph nodes and systemic metastases to bone, brain, left lung and kidney. This pattern closely simulates the clinical behavior of NSCLC. In this model both the primary lung tumor and metastatic tumor in bone and left lung express MMP-2, MT1MMP, and TIMP-2, with the metastatic lesions tending to have a higher level of expression. Zymographic analysis revealed high levels of proMMP-2 expression in beth primary and metastatic tumors. MMP-2, MT1-MMP, and TIMP-2 may play a critical role in tumor invasion and metastasis in these models. Prinomastat (AG3340), a novel synthetic MMP inhibitor mainly targeting gelatinases (MMP-2 and -9), and MT1-MMP, was tested in our H460 metastatic orthotopic lung cancer model. Prinomastat (AG3340) (100 mg/kg po bid) in combination with carboplatin (10 mg/kg ip weekly) significantly prolonged survival compared to either drug alone. Toxicity was minimal and tolerance much better than cytotoxic agents tested in this model. These orthotopic lung cancer models may be useful for studying complex interaction of MMPs and TIMPs in vivo. Particularly, the H460 orthotopic model that metastasizes widely may be an effective preclinical screening tool to assess the efficacy of novel
203
MMP inhibitors specific for the MMPs and TIMP expressed in these tumors.
~ - - ~ Increasing serum levels of basic fibroblast growth factor in non small cell lung cancer indicates progressive disease D. Brattstr6m 1, M. Bergqvist 1, P. Hesselius 1, A. Larsson 2, G. Wagenius 3, O. Brodin 3. 1Department of Oncology, Akademiska
Hospital, Uppsala Universi~ 751 85 Uppsala; 2Department of Clinical Chemistry, Akademiska Hospital, Uppsala University, 751 85 Uppsala; 3Department of Oncology, Uppsala Akademiska Hospital, Uppsala, Sweden Basic fibroblast growth factor (bFGF) is a mitogen with many properties; one of them is to stimulate angiogenesis in a very potent manner. In vitro and in vivo studies have implicated that bFGF and VEGF are closely correlated and act synergistically in inducing angiogenesis, yet maybe in different temporal fashion. Our earlier study on S-bFGF indicates that S-bFGF have prognostic information of great importance for operable non-small cell lung cancer (NSCLC). In the present study we focused on the impact of alterations of S-bFGF during treatment in patients with inoperable NSCLC. We analysed 486 serum samples from 72 patients with inoperable NSCLC for bFGF values. For the detection of S-bFGF we used a commercially available immunosorbent assay (R&D Systems Inc., Minneapolis, MN, USA). All patients gave informed consent prior collection and a minimum of three samples were collected during the course of treatment. Thirty-eight patients were staged as Ilia, 24 patients as IIIb and 10 patients as stage IV. Mean value of S-bFGF in all samples (no 486) was 9.6 pg/ml (confidence interval (CI) 8.6-10.6). Mean values of S-bFGF in samples stratified into stage were in Ilia 8.5 pg/ml (CI 5.8-11.1), stage IIIb 11.8 pg/ml (CI 6.3-17.4) and for stage IV 12.0 pg/ml (CI 5.3-18.6). Regression analysis using a time-index statistical model demonstrated a significant correlation between increasing values of S-bFGF and date of death (p < 0.005). Regression analysis also demonstrated a significant correlation between platelets and bFGF, e.g. with each rise of 1 x 109 in platelet count, S-bFGF rises 0.021 pg/ml (p < 0.0001). When analysing location of metastases and S-bFGF values, no differences could be demonstrated between brain and skeleton compared to no sign of metastases. But, when multiple metastases and/or liver metastases were present there was a difference between S-bFGF (mean 14.7 pg/ml CI 10.4-18.9) compared to no sign of metastases (mean 9.2 pg/ml CI 8.2-10.3).
[-6--9-~ HER-2 status and response to chemotherapy in non small lung cancer J.F. Morere, M. Kambouchner, M.C. Pailler, S. Piperno-Neumann, E. Goguel, A. Martin, J.L. Braau. Avicenne Hospital, Bobigny, France Background: HER-2/neu gene overexpression may be important in cancer progression. Numerous studies have investigated the relationship between HER2 status and prognosis or response to therapies mostly in breast cancer. Methods: In this retrospective study we have evaluated the correlation of HER-2 status and the response to chemotherapy in 50 patients with advanced adenocarcinoma or large cell lung cancer. Bronchial biopsies were obtained by fibroscopy or surgery. Paraffin embedded tumor tissue were stained with an antibody to a site of the internal domain of the HER-2 receptor (CB11). Results: Overall response rate to standard chemotherapy in the whole population was 26%. Fourty three biopsies could be stained. Weak positive staining was observed in 16 out of 43 (37%). No statistically significant difference was observed in response to chemotherapy according to HER-2 status. Conclusion: In our population of patients, HER-2 staining with CB11 does not correlate with response to chemotherapy. A prospective study combining immunohistochemistry, and fluorescence in situhybridisation is ongoing.