Extended-stage small cell lung cancer (SCLC) responds to platinum/vepeside-based first-line chemotherapy but relapses rapidly as drug-resistant tumor. Topotecan (TPT) is the single chemotherapeutic agent approved for ...Extended-stage small cell lung cancer (SCLC) responds to platinum/vepeside-based first-line chemotherapy but relapses rapidly as drug-resistant tumor. Topotecan (TPT) is the single chemotherapeutic agent approved for second-line treatment of SCLC. However, the response to TPT is short-lived and novel treatment modalities need to be developed. Sequential treatment of cytotoxic drugs and inhibitors of cyclin-dependent kinases (CDKs) showed promising preclinical anticancer activity and, in the present work, combinations of TPT with CDK inhibitors olomoucine, roscovitine and CDK4I are shown to exhibit synergistic cytotoxic activity against SCLC cell lines. Highest activity was found against TPT-resistant NCI-H417 and DMS153 cell lines and moderate chemosensitizing effects against a primary SCLC cell line and sensitive GLC19 cells at levels of CDK inhibitors which exerted low toxicity. A combination of 0.6 μM TPT with 0.6 μM roscovitine, exhibiting no significant cytotoxicity as single agents, reduced viability of the TPT-resistant NCI-H417 line (IC50 > 10 μM) by 50%. In the TPT resistant cell lines olomoucine and roscovitine, targeting CDK1,2,5,7, were highly effective, whereas in the more sensitive cell lines CDK4I, inhibiting mainly CDK4/6, showed activity. In NCI-417 cells, preincubation with roscovitine for one day proved synergistic with TPT. Thus, in good accordance with previous findings, CDK inhibitors are able to convert SCLC cancer cells which are cell-cycle arrested by a blockade of topoisomerase I by TPT to apoptotic cells. Since nowadays several CDK inhibitors are at various phases of clinical testing their combination with TPT seems to constitute a promising approach to improve second-line chemotherapy in SCLC.展开更多
目的:探讨Topotecan在结直肠癌手术后辅助化疗的效果。方法:2003年6月￣2005年6月收治结直肠癌患者212例,纳入观察对象58例。男39例,女19例,年龄32￣75岁,平均56.6岁。癌胚抗原(CEA)阳性46例,其中结肠癌分期Dukes A 11例,Dukes B 17例,D...目的:探讨Topotecan在结直肠癌手术后辅助化疗的效果。方法:2003年6月￣2005年6月收治结直肠癌患者212例,纳入观察对象58例。男39例,女19例,年龄32￣75岁,平均56.6岁。癌胚抗原(CEA)阳性46例,其中结肠癌分期Dukes A 11例,Dukes B 17例,Dukes C 9例共37例;直肠癌分期Dukes A 4例,Dukes B 4例,Dukes C 1例共9例。伴有肝转移或者髂血管旁淋巴结转移侵犯而不能切除者12例,其中结肠癌5例,直肠癌7例。在肝转移或者髂血管旁淋巴结转移侵犯而不能切除的病例构成中结肠癌5例其中肝内转移灶直径≥2.0cm单灶3例,多灶2例。直肠癌7例中肝内转移3例直径≥2.0cm均为单灶,髂血管旁淋巴结转移侵犯而不能切除,术前B超检查直径>2.0cm4例。采用性别、年龄以及CEA阳性、远处转移配对分为治疗组与对照组,治疗组:Topotecan1.25mg/m2静脉滴入,d1、3、5,Q21￣28d加替加氟0.6g静脉滴入d1￣5,Q21￣28d;对照组丝裂霉素2.5mg/m2静脉滴入,d14,Q21￣28d加替加氟0.6g静脉滴入d1￣5,Q21￣28d。随访3个月后分别对CEA水平、转移灶大小进行评价。结果:治疗组与对照组疗效对比分别是完全缓解(CR)68.9%vs48.2%。总有效率分别为86.2%vs68.9%组间比较P>0.05。结论:结直肠癌手术后应用Topotecan联合方案可以较好地控制肿瘤发展,降低CEA水平,使转移灶缩小或部分消失。与丝裂霉素一样可用于结直肠癌手术后辅助性化疗。展开更多
Topotecan (TPT), a semisynthetic analogue of the natural product camptothecin is a cell cycle-specific drug with antitumor activity. To clarify the effect of TPT on SUD4 and DOHH2 cell line in this study, we examined ...Topotecan (TPT), a semisynthetic analogue of the natural product camptothecin is a cell cycle-specific drug with antitumor activity. To clarify the effect of TPT on SUD4 and DOHH2 cell line in this study, we examined the apoptosis and cell cycle changes of the two human cancer cell lines by exposing to TPT for 18 hours at various concentrations. The linear relationship between apoptosis cell number and the concentration of TPT was observed by means of Flow Cytometry and Annexin V assay. Then, DOHH2 cell is much more sensitive to TPT than SUD4 cell. In addition, Cell Question Software Assay showed positive relationship between the frequency of cells accumulated in S-phase and the concentration of TPT. The least concentration of TPT to change cell cycle is 5 nmol·L?1 in both cell lines. These results suggest that the inducing apoptosis of cancer cells is one of mechanism of TPT antitumor activity.展开更多
Objective: To observe the synergistic efficacy between Adenovirus-mediated bcl-Xs(Adv-bcl-Xs) gene transfer and chemotherapy on ovarian cancer cell growth. Methods: NuTu-19 cells were infected by different titers of A...Objective: To observe the synergistic efficacy between Adenovirus-mediated bcl-Xs(Adv-bcl-Xs) gene transfer and chemotherapy on ovarian cancer cell growth. Methods: NuTu-19 cells were infected by different titers of Adv-bcl-Xs and treated with topotecan in the meantime. Cell proliferation was measured 3 days later by MTT. Graphical representations and statistical analyses for their interaction in tumor cells were done. Results: The statistical result and Graphical representations of the statistical modeling showed synergy effect on cell growth inhibition (P<0.01). Conclusion: There were synergistic efficacies between Adv-bcl-Xs gene therapy and Topotecan in ovarian cancer cell growth.展开更多
Background: Prostate cancer is the most common cancer in men over the age of 60 in Western countries. An estimated 241,740 new cases of prostate cancer have been diagnosed in the United States in 2012 with a death tol...Background: Prostate cancer is the most common cancer in men over the age of 60 in Western countries. An estimated 241,740 new cases of prostate cancer have been diagnosed in the United States in 2012 with a death toll of 28,170. Varieties of natural phytochemicals such as genistein and topotecan have shown potential chemotherapeutic capacities and are being used to inhibit the growth and proliferation of cell in prostate cancer. Purpose of Study: In this study, we aim to determine the efficacy of Vitamin D3-Topotecan combination compared to Genistein-Topotecan in apoptosis induction in LNCaP prostate cancer cells. Methods: LNCaP cells were grown in complete RPMI medium and cultured at 37°C, 5% CO2 for 23 - 48 hrs to achieve 70% - 80% confluence. The cells were then treated with Genistein-Topotecan, Vitamin D3-Topotecan combination and TPT alone for 24 - 48 hours. In addition, post-treatment assayed using: Trypan Blue exclusion and MTT for cell viability, Ethidium bromide/Acridine orange to determine apoptosis induction, Rhodamine 123/Ethidium bromide to differentiate between viable, apoptotic, and necrotic cells, as well as to assess possible apoptotic mechanism, and DNA fragmentation to discriminate between apoptotic and necrotic cell death. Results: The overall data indicated the dose-and time-dependent cell death in the LNCaP cells and apoptosis as the major mechanism of treatment-induced cell growth arrest. Conclusion: The Genistein-Topotecan combination treatment was significantly more efficacious in growth inhibition of LNCaP cells compared to Vitamin D3-Topotecan or Topotecan alone.展开更多
Objective: Cervical cancer represents the third most commonly diagnosed cancer and is an important cause of death for women suffering with malignancies. Patients who are refractory or progressed after first-line palli...Objective: Cervical cancer represents the third most commonly diagnosed cancer and is an important cause of death for women suffering with malignancies. Patients who are refractory or progressed after first-line palliative treatment have a dismal prognosis and no second-line chemotherapy is considered standard so far. Several agents have been investigated in this setting and topotecan is one of the most characterized. The objective of this study was to evaluate response rate (RR), progression-free survival (PFS), overall survival (OS) and toxicity of topotecan in second palliative line for cervical cancer. Methods: An analysis was performed of all patients with recurrent or metastatic cervical cancer treated with topotecan in second palliative line at Brazilian National Cancer Institute, between 2008 and 2010. Results: A total of 73 courses of topotecan were given in the current study (median: 3.5 cycles;range 1 - 6). Anemia was the most frequent adverse event (grade 2:35%;grade 3:30%). Of the 20 patients evaluable, there were 2 partial responders to the treatment. The overall response rate (ORR) was 10%;3 patients (15%) had stable disease as maximum response. The median PFS for the entire group was 2.93 months (95% CI 2.41 - 3.45) and OS was 4.66 months (95% CI 1.21 - 8.11). Conclusion: The limited activity of topotecan schemas in second-line treatment of cervical cancer and the associated overall toxicity may not justify their use in this setting. Patients who progress after first-line treatment may be offered participation in clinical trials, other second-line agents or best supportive care measures.展开更多
Background: Artemisinin dimer oxime – dimer molecule synthesized from artemisinin possesses high bioavailability and marked in vitro anticancer activities against solid tumor?derived cell lines, endothelial cell prol...Background: Artemisinin dimer oxime – dimer molecule synthesized from artemisinin possesses high bioavailability and marked in vitro anticancer activities against solid tumor?derived cell lines, endothelial cell proliferation, migration, and angiogenic processes. Numerous murine models have been developed to study human cancer. The most widely used models are the human tumor xenograft mouse model. Materials and Methods: In this study, human tumor cells(NCI?H640, 1 × 107 in 100 μL) are implanted subcutaneously, or 1 × 107 in 50 μL in the thoracic cavity, in athymic nude mice(nu/nu). The implanted cells were allowed to grow for 10 days before initiation of drug treatment(dimer oxime and topotecan, ip). Tumor volume and thoracic/body weight ratio were recorded. Results: We successfully established subcutaneous and thoracic xenografts with human nonsmall cell lung cancer cell line xenografts in athymic nude mice in only 10 days. Using these models, we attempted treatment of xenografts with topotecan – a known anticancer drug and artemisinin dimer oxime or combination of these two drugs. Combination therapy showed a significant reduction in tumor volume and tumor/body weight. Treatments with combination of topotecan and dimer oxime resulted in the reduced mortality rates in comparison with untreated mice. Conclusions: Xenograft tumor models are useful for preclinical screening of new pharmacophores. From this preliminary study, it appears that combination of dimer oxime and topotecan may be used as chemotherapeutic agents against nonsmall cell lung cancer. Further studies are needed to evaluate other combination treatment regimens as well as the mechanism(s) of action.展开更多
Small cell lung cancer(SCLC)is frequently disseminated and has a dismal prognosis with survival times of approximately two years.This cancer responds well to initial chemotherapy but recurs within a short time as a gl...Small cell lung cancer(SCLC)is frequently disseminated and has a dismal prognosis with survival times of approximately two years.This cancer responds well to initial chemotherapy but recurs within a short time as a globally chemoresistant tumor.Circulating tumor cells(CTCs)are held responsible for metastasis,the extremely high numbers of these cells in advanced SCLC allowed us to establish several permanent CTC cell lines.These CTCs are distinguished by the spontaneous formation of large spheroids,termed tumorospheres,in regular tissue culture.These contain quiescent and hypoxic cells in their interior and are associated with high chemoresistance compared to single cell cultures.Nine CTC lines were compared for their expression of 84 proteins associated with cancer either as single cells or in the form of tumorospheres in Western blot arrays.With the exception of the UHGc5 line,all other CTC lines express EpCAM and lack a complete EpCAM-negative,vimentin-positive epithelial-mesenchymal transition(EMT)phenotype.Upon formation of tumorospheres the expression of EpCAM,that mediates cell-cell adhesion is markedly upregulated.Proteins such as E-Cadherin,p27 KIP1,Progranulin,BXclx,Galectin-3,and Survivin showed variable changes for the distinct CTC cell lines.In conclusion,EpCAM presents the most critical marker for individual SCLC CTCs and the assembly of highly chemoresistant tumorospheres.展开更多
文摘Extended-stage small cell lung cancer (SCLC) responds to platinum/vepeside-based first-line chemotherapy but relapses rapidly as drug-resistant tumor. Topotecan (TPT) is the single chemotherapeutic agent approved for second-line treatment of SCLC. However, the response to TPT is short-lived and novel treatment modalities need to be developed. Sequential treatment of cytotoxic drugs and inhibitors of cyclin-dependent kinases (CDKs) showed promising preclinical anticancer activity and, in the present work, combinations of TPT with CDK inhibitors olomoucine, roscovitine and CDK4I are shown to exhibit synergistic cytotoxic activity against SCLC cell lines. Highest activity was found against TPT-resistant NCI-H417 and DMS153 cell lines and moderate chemosensitizing effects against a primary SCLC cell line and sensitive GLC19 cells at levels of CDK inhibitors which exerted low toxicity. A combination of 0.6 μM TPT with 0.6 μM roscovitine, exhibiting no significant cytotoxicity as single agents, reduced viability of the TPT-resistant NCI-H417 line (IC50 > 10 μM) by 50%. In the TPT resistant cell lines olomoucine and roscovitine, targeting CDK1,2,5,7, were highly effective, whereas in the more sensitive cell lines CDK4I, inhibiting mainly CDK4/6, showed activity. In NCI-417 cells, preincubation with roscovitine for one day proved synergistic with TPT. Thus, in good accordance with previous findings, CDK inhibitors are able to convert SCLC cancer cells which are cell-cycle arrested by a blockade of topoisomerase I by TPT to apoptotic cells. Since nowadays several CDK inhibitors are at various phases of clinical testing their combination with TPT seems to constitute a promising approach to improve second-line chemotherapy in SCLC.
文摘目的:探讨Topotecan在结直肠癌手术后辅助化疗的效果。方法:2003年6月￣2005年6月收治结直肠癌患者212例,纳入观察对象58例。男39例,女19例,年龄32￣75岁,平均56.6岁。癌胚抗原(CEA)阳性46例,其中结肠癌分期Dukes A 11例,Dukes B 17例,Dukes C 9例共37例;直肠癌分期Dukes A 4例,Dukes B 4例,Dukes C 1例共9例。伴有肝转移或者髂血管旁淋巴结转移侵犯而不能切除者12例,其中结肠癌5例,直肠癌7例。在肝转移或者髂血管旁淋巴结转移侵犯而不能切除的病例构成中结肠癌5例其中肝内转移灶直径≥2.0cm单灶3例,多灶2例。直肠癌7例中肝内转移3例直径≥2.0cm均为单灶,髂血管旁淋巴结转移侵犯而不能切除,术前B超检查直径>2.0cm4例。采用性别、年龄以及CEA阳性、远处转移配对分为治疗组与对照组,治疗组:Topotecan1.25mg/m2静脉滴入,d1、3、5,Q21￣28d加替加氟0.6g静脉滴入d1￣5,Q21￣28d;对照组丝裂霉素2.5mg/m2静脉滴入,d14,Q21￣28d加替加氟0.6g静脉滴入d1￣5,Q21￣28d。随访3个月后分别对CEA水平、转移灶大小进行评价。结果:治疗组与对照组疗效对比分别是完全缓解(CR)68.9%vs48.2%。总有效率分别为86.2%vs68.9%组间比较P>0.05。结论:结直肠癌手术后应用Topotecan联合方案可以较好地控制肿瘤发展,降低CEA水平,使转移灶缩小或部分消失。与丝裂霉素一样可用于结直肠癌手术后辅助性化疗。
文摘Topotecan (TPT), a semisynthetic analogue of the natural product camptothecin is a cell cycle-specific drug with antitumor activity. To clarify the effect of TPT on SUD4 and DOHH2 cell line in this study, we examined the apoptosis and cell cycle changes of the two human cancer cell lines by exposing to TPT for 18 hours at various concentrations. The linear relationship between apoptosis cell number and the concentration of TPT was observed by means of Flow Cytometry and Annexin V assay. Then, DOHH2 cell is much more sensitive to TPT than SUD4 cell. In addition, Cell Question Software Assay showed positive relationship between the frequency of cells accumulated in S-phase and the concentration of TPT. The least concentration of TPT to change cell cycle is 5 nmol·L?1 in both cell lines. These results suggest that the inducing apoptosis of cancer cells is one of mechanism of TPT antitumor activity.
基金This work was supported by a grant from the Science Foundation of National Committee of Education of China (No. 96361017).
文摘Objective: To observe the synergistic efficacy between Adenovirus-mediated bcl-Xs(Adv-bcl-Xs) gene transfer and chemotherapy on ovarian cancer cell growth. Methods: NuTu-19 cells were infected by different titers of Adv-bcl-Xs and treated with topotecan in the meantime. Cell proliferation was measured 3 days later by MTT. Graphical representations and statistical analyses for their interaction in tumor cells were done. Results: The statistical result and Graphical representations of the statistical modeling showed synergy effect on cell growth inhibition (P<0.01). Conclusion: There were synergistic efficacies between Adv-bcl-Xs gene therapy and Topotecan in ovarian cancer cell growth.
文摘Background: Prostate cancer is the most common cancer in men over the age of 60 in Western countries. An estimated 241,740 new cases of prostate cancer have been diagnosed in the United States in 2012 with a death toll of 28,170. Varieties of natural phytochemicals such as genistein and topotecan have shown potential chemotherapeutic capacities and are being used to inhibit the growth and proliferation of cell in prostate cancer. Purpose of Study: In this study, we aim to determine the efficacy of Vitamin D3-Topotecan combination compared to Genistein-Topotecan in apoptosis induction in LNCaP prostate cancer cells. Methods: LNCaP cells were grown in complete RPMI medium and cultured at 37°C, 5% CO2 for 23 - 48 hrs to achieve 70% - 80% confluence. The cells were then treated with Genistein-Topotecan, Vitamin D3-Topotecan combination and TPT alone for 24 - 48 hours. In addition, post-treatment assayed using: Trypan Blue exclusion and MTT for cell viability, Ethidium bromide/Acridine orange to determine apoptosis induction, Rhodamine 123/Ethidium bromide to differentiate between viable, apoptotic, and necrotic cells, as well as to assess possible apoptotic mechanism, and DNA fragmentation to discriminate between apoptotic and necrotic cell death. Results: The overall data indicated the dose-and time-dependent cell death in the LNCaP cells and apoptosis as the major mechanism of treatment-induced cell growth arrest. Conclusion: The Genistein-Topotecan combination treatment was significantly more efficacious in growth inhibition of LNCaP cells compared to Vitamin D3-Topotecan or Topotecan alone.
文摘Objective: Cervical cancer represents the third most commonly diagnosed cancer and is an important cause of death for women suffering with malignancies. Patients who are refractory or progressed after first-line palliative treatment have a dismal prognosis and no second-line chemotherapy is considered standard so far. Several agents have been investigated in this setting and topotecan is one of the most characterized. The objective of this study was to evaluate response rate (RR), progression-free survival (PFS), overall survival (OS) and toxicity of topotecan in second palliative line for cervical cancer. Methods: An analysis was performed of all patients with recurrent or metastatic cervical cancer treated with topotecan in second palliative line at Brazilian National Cancer Institute, between 2008 and 2010. Results: A total of 73 courses of topotecan were given in the current study (median: 3.5 cycles;range 1 - 6). Anemia was the most frequent adverse event (grade 2:35%;grade 3:30%). Of the 20 patients evaluable, there were 2 partial responders to the treatment. The overall response rate (ORR) was 10%;3 patients (15%) had stable disease as maximum response. The median PFS for the entire group was 2.93 months (95% CI 2.41 - 3.45) and OS was 4.66 months (95% CI 1.21 - 8.11). Conclusion: The limited activity of topotecan schemas in second-line treatment of cervical cancer and the associated overall toxicity may not justify their use in this setting. Patients who progress after first-line treatment may be offered participation in clinical trials, other second-line agents or best supportive care measures.
文摘Background: Artemisinin dimer oxime – dimer molecule synthesized from artemisinin possesses high bioavailability and marked in vitro anticancer activities against solid tumor?derived cell lines, endothelial cell proliferation, migration, and angiogenic processes. Numerous murine models have been developed to study human cancer. The most widely used models are the human tumor xenograft mouse model. Materials and Methods: In this study, human tumor cells(NCI?H640, 1 × 107 in 100 μL) are implanted subcutaneously, or 1 × 107 in 50 μL in the thoracic cavity, in athymic nude mice(nu/nu). The implanted cells were allowed to grow for 10 days before initiation of drug treatment(dimer oxime and topotecan, ip). Tumor volume and thoracic/body weight ratio were recorded. Results: We successfully established subcutaneous and thoracic xenografts with human nonsmall cell lung cancer cell line xenografts in athymic nude mice in only 10 days. Using these models, we attempted treatment of xenografts with topotecan – a known anticancer drug and artemisinin dimer oxime or combination of these two drugs. Combination therapy showed a significant reduction in tumor volume and tumor/body weight. Treatments with combination of topotecan and dimer oxime resulted in the reduced mortality rates in comparison with untreated mice. Conclusions: Xenograft tumor models are useful for preclinical screening of new pharmacophores. From this preliminary study, it appears that combination of dimer oxime and topotecan may be used as chemotherapeutic agents against nonsmall cell lung cancer. Further studies are needed to evaluate other combination treatment regimens as well as the mechanism(s) of action.
基金partially funded by the“Medical Scientific Fund of the Mayor of the City of Vienna”,Grant Number 21040 to Dr.C.Lang.
文摘Small cell lung cancer(SCLC)is frequently disseminated and has a dismal prognosis with survival times of approximately two years.This cancer responds well to initial chemotherapy but recurs within a short time as a globally chemoresistant tumor.Circulating tumor cells(CTCs)are held responsible for metastasis,the extremely high numbers of these cells in advanced SCLC allowed us to establish several permanent CTC cell lines.These CTCs are distinguished by the spontaneous formation of large spheroids,termed tumorospheres,in regular tissue culture.These contain quiescent and hypoxic cells in their interior and are associated with high chemoresistance compared to single cell cultures.Nine CTC lines were compared for their expression of 84 proteins associated with cancer either as single cells or in the form of tumorospheres in Western blot arrays.With the exception of the UHGc5 line,all other CTC lines express EpCAM and lack a complete EpCAM-negative,vimentin-positive epithelial-mesenchymal transition(EMT)phenotype.Upon formation of tumorospheres the expression of EpCAM,that mediates cell-cell adhesion is markedly upregulated.Proteins such as E-Cadherin,p27 KIP1,Progranulin,BXclx,Galectin-3,and Survivin showed variable changes for the distinct CTC cell lines.In conclusion,EpCAM presents the most critical marker for individual SCLC CTCs and the assembly of highly chemoresistant tumorospheres.
