AIM:To investigate the antiproliferative activity of cinobufacini on human hepatocellular carcinoma HepG2 cells and the possible mechanism of its action.METHODS:HepG2 cells were treated with different concentrations o...AIM:To investigate the antiproliferative activity of cinobufacini on human hepatocellular carcinoma HepG2 cells and the possible mechanism of its action.METHODS:HepG2 cells were treated with different concentrations of cinobufacini.Cell viability was measured by methylthiazolyl tetrazolium(MTT) assay.Cell cycledistribution was analyzed by flow cytometry(FCM).Cytoskeletal and nuclear alterations were observed by fluorescein isothiocyanate-phalloidin and DAPI staining under a laser scanning confocal microscope.Changes in morphology and ultrastructure of cells were detected by atomic force microscopy(AFM) at the nanoscale level.RESULTS:MTT assay indicated that cinobufacini significantly inhibited the viability of HepG2 cells in a dosedependent manner.With the concentration of cinobufacini increasing from 0 to 0.10 mg/m L,the cell viability decreased from 74.9% ± 2.7% to 49.41% ± 2.2% and 39.24% ± 2.1%(P < 0.05).FCM analysis demonstrated cell cycle arrest at S phase induced by cinobufacini.The immunofluorescence studies of cytoskeletal and nuclear morphology showed that after cinobufacini treatment,the regular reorganization of actin filaments in HepG2 cells become chaotic,while the nuclei were not damaged seriously.Additionally,high-resolution AFM imaging revealed that cell morphology and ultrastructure changed a lot after treatment with cinobufacini.It appeared as significant shrinkage and deep pores in the cell membrane,with larger particles and a rougher cell surface.CONCLUSION:Cinobufacini inhibits the viability of HepG2 cells via cytoskeletal destruction and cell membrane toxicity.展开更多
Chinese medicine cinobufacini is an extract from the dried skin of Bufo bufo gargarizans Cantor,with active ingredients of bufadienolides and indole alkaloids.With further research and clinical applications,it is foun...Chinese medicine cinobufacini is an extract from the dried skin of Bufo bufo gargarizans Cantor,with active ingredients of bufadienolides and indole alkaloids.With further research and clinical applications,it is found that cinobufacini alone or in combination with other therapeutic methods can play an anti-tumor role by controlling proliferation of tumor cells,promoting apoptosis,inhibiting formation of tumor neovascularization,reversing multidrug resistance,and regulating immune response;it also has the functions of relieving cancer pain and regulating immune function.In this paper,the chemical composition,pharmacological effects,clinical applications,and adverse reactions of cinobufacini are summarized.However,the extraction of monomer components of cinobufacini,the relationship between different mechanisms,and the causes of adverse reactions need to be further studied.Also,high-quality clinical studies should be conducted.展开更多
Objective： To investigate the therapeutic effects of Jiedu Granules （解毒颗粒）, a Chinese medicine （CM） compound, plus Cinobufacini Injection （华蟾素注射液）, which was extracted from skin of Bufo bufo gargariza...Objective： To investigate the therapeutic effects of Jiedu Granules （解毒颗粒）, a Chinese medicine （CM） compound, plus Cinobufacini Injection （华蟾素注射液）, which was extracted from skin of Bufo bufo gargarizans Cantor, to prevent the recurrence of hepatocellular carcinoma （HCC） after surgical resection. Methods： In this case-control trial, a total of 120 patients who stayed in Changhai Hospital were enrolled from December 2001 to December 2006. Sixty patients were treated with Jiedu Granules plus Cinobufacini Injection to prevent tumor recurrence after operation （CM group） and 60 patients were treated with transcatheter arterial chemoembolization （TACE） after operation （TACE group）. Progression-free survival （PFS） and overall survival （OS） rates were determined to evaluate the therapeutic effects of post-operative management of patients with HCC. Results： PFS in the CM group was 18.07 months [95% confidence interval （CI）： 12.49-23.65] and the 1-, 2-, 3-, 4- and 5-year PFS rates were 61%, 39%, 26%, 22% and 12%, respectively. PFS in the TACE group was 8.03 months （95% CI： 6.63-9.44） and the 1-, 2-, 3-, 4- and 5-year PFS rates were 34%, 11%, 7%, 2% and 0%, respectively. There was significant difference in survival rate between the two groups （P〈0.01）. The mean survival time （MST） of patients in the CM group was 49.53 months versus 39.90 months of the TACE group. The 1-, 2-, 3-, 4- and 5-year survival rates were 90%, 82%, 80%, 70% and 63%, respectively, in the CM group, and 79%, 70%, 60%, 60% and 36%, respectively, in the TACE group. There was significant difference in survival time between the two groups （P=0.045）. Conclusions： Jiedu Granules plus Cinobufacini Injection, a combination that is commonly used for post-operation management of HCC, can postpone tumor recurrence and metastasis, prolong the survival time and increase the survival rate of post-surgical patients with HCC. However, these findings need to be confirmed in a prospective, randomized controlled trial.展开更多
To investigate the mechanism of the anti-tumor activity of cinobufacini on the breast cancer cell line T-47D,the inhibitory effect of cinobufacini on the proliferation of T-47D was detected via MTT assay and the morph...To investigate the mechanism of the anti-tumor activity of cinobufacini on the breast cancer cell line T-47D,the inhibitory effect of cinobufacini on the proliferation of T-47D was detected via MTT assay and the morphological changes of T-47D and HBL-100 cells caused by cinobufacini were observed with an inverted microscope.Cell apoptosis and cell cycle stages were detected by flow cytometry analysis.The effects of cinobufacini on the expression of active-form and pro-form of caspase-3 were assessed by Western blot analysis.Cinobufacini dramatically inhibited T-47D proliferation in a dose-and time-dependent manner.We found that more than 20％ of T-47D cells were killed after treatment with 20 mg/mL cinobufacini for 24 h in vitro.After 6 d of treatment with 20 mg/mL cinobufacini,the cell survival rate decreased by more than 40％.Flow cytometric analysis demonstrated that cinobufacini induced significant apoptosis and changes of the cell cycle distribution of T-47D cells.We used breast cell line HBL-100 as the control,the above experiments except cell cycle analysis showed that cinobufacini more obviously induced the apoptosis of T-47D cells than that of HBL-100 cells.Western blot analysis confirmed the protein expression of active caspase-3 increased with increasing the dose of cinobufacini.These results indicate that cinobufacini induces the apoptosis of T-47D cells via the up-regulation of caspase-3.展开更多
文摘AIM:To investigate the antiproliferative activity of cinobufacini on human hepatocellular carcinoma HepG2 cells and the possible mechanism of its action.METHODS:HepG2 cells were treated with different concentrations of cinobufacini.Cell viability was measured by methylthiazolyl tetrazolium(MTT) assay.Cell cycledistribution was analyzed by flow cytometry(FCM).Cytoskeletal and nuclear alterations were observed by fluorescein isothiocyanate-phalloidin and DAPI staining under a laser scanning confocal microscope.Changes in morphology and ultrastructure of cells were detected by atomic force microscopy(AFM) at the nanoscale level.RESULTS:MTT assay indicated that cinobufacini significantly inhibited the viability of HepG2 cells in a dosedependent manner.With the concentration of cinobufacini increasing from 0 to 0.10 mg/m L,the cell viability decreased from 74.9% ± 2.7% to 49.41% ± 2.2% and 39.24% ± 2.1%(P < 0.05).FCM analysis demonstrated cell cycle arrest at S phase induced by cinobufacini.The immunofluorescence studies of cytoskeletal and nuclear morphology showed that after cinobufacini treatment,the regular reorganization of actin filaments in HepG2 cells become chaotic,while the nuclei were not damaged seriously.Additionally,high-resolution AFM imaging revealed that cell morphology and ultrastructure changed a lot after treatment with cinobufacini.It appeared as significant shrinkage and deep pores in the cell membrane,with larger particles and a rougher cell surface.CONCLUSION:Cinobufacini inhibits the viability of HepG2 cells via cytoskeletal destruction and cell membrane toxicity.
基金Supported by the National Natural Science Foundation of China(No.82274084)Inheritance and Innovation of Traditional Chinese Medicine and Key Scientific Research Project of"Qin Medicine"Development of Shaanxi Administration of Traditional Chinese Medicine(No.2021-02-22-007)Subject Innovation Team of Shaanxi University of Chinese Medicine(No.2019-YL10)。
文摘Chinese medicine cinobufacini is an extract from the dried skin of Bufo bufo gargarizans Cantor,with active ingredients of bufadienolides and indole alkaloids.With further research and clinical applications,it is found that cinobufacini alone or in combination with other therapeutic methods can play an anti-tumor role by controlling proliferation of tumor cells,promoting apoptosis,inhibiting formation of tumor neovascularization,reversing multidrug resistance,and regulating immune response;it also has the functions of relieving cancer pain and regulating immune function.In this paper,the chemical composition,pharmacological effects,clinical applications,and adverse reactions of cinobufacini are summarized.However,the extraction of monomer components of cinobufacini,the relationship between different mechanisms,and the causes of adverse reactions need to be further studied.Also,high-quality clinical studies should be conducted.
基金Supported by National Key Technology R&D Program for the 11th Five-Year Plan(No.2006 BAI04A06)
文摘Objective： To investigate the therapeutic effects of Jiedu Granules （解毒颗粒）, a Chinese medicine （CM） compound, plus Cinobufacini Injection （华蟾素注射液）, which was extracted from skin of Bufo bufo gargarizans Cantor, to prevent the recurrence of hepatocellular carcinoma （HCC） after surgical resection. Methods： In this case-control trial, a total of 120 patients who stayed in Changhai Hospital were enrolled from December 2001 to December 2006. Sixty patients were treated with Jiedu Granules plus Cinobufacini Injection to prevent tumor recurrence after operation （CM group） and 60 patients were treated with transcatheter arterial chemoembolization （TACE） after operation （TACE group）. Progression-free survival （PFS） and overall survival （OS） rates were determined to evaluate the therapeutic effects of post-operative management of patients with HCC. Results： PFS in the CM group was 18.07 months [95% confidence interval （CI）： 12.49-23.65] and the 1-, 2-, 3-, 4- and 5-year PFS rates were 61%, 39%, 26%, 22% and 12%, respectively. PFS in the TACE group was 8.03 months （95% CI： 6.63-9.44） and the 1-, 2-, 3-, 4- and 5-year PFS rates were 34%, 11%, 7%, 2% and 0%, respectively. There was significant difference in survival rate between the two groups （P〈0.01）. The mean survival time （MST） of patients in the CM group was 49.53 months versus 39.90 months of the TACE group. The 1-, 2-, 3-, 4- and 5-year survival rates were 90%, 82%, 80%, 70% and 63%, respectively, in the CM group, and 79%, 70%, 60%, 60% and 36%, respectively, in the TACE group. There was significant difference in survival time between the two groups （P=0.045）. Conclusions： Jiedu Granules plus Cinobufacini Injection, a combination that is commonly used for post-operation management of HCC, can postpone tumor recurrence and metastasis, prolong the survival time and increase the survival rate of post-surgical patients with HCC. However, these findings need to be confirmed in a prospective, randomized controlled trial.
基金Supported by the National Natural Science Foundation of China（No.30300336）.
文摘To investigate the mechanism of the anti-tumor activity of cinobufacini on the breast cancer cell line T-47D,the inhibitory effect of cinobufacini on the proliferation of T-47D was detected via MTT assay and the morphological changes of T-47D and HBL-100 cells caused by cinobufacini were observed with an inverted microscope.Cell apoptosis and cell cycle stages were detected by flow cytometry analysis.The effects of cinobufacini on the expression of active-form and pro-form of caspase-3 were assessed by Western blot analysis.Cinobufacini dramatically inhibited T-47D proliferation in a dose-and time-dependent manner.We found that more than 20％ of T-47D cells were killed after treatment with 20 mg/mL cinobufacini for 24 h in vitro.After 6 d of treatment with 20 mg/mL cinobufacini,the cell survival rate decreased by more than 40％.Flow cytometric analysis demonstrated that cinobufacini induced significant apoptosis and changes of the cell cycle distribution of T-47D cells.We used breast cell line HBL-100 as the control,the above experiments except cell cycle analysis showed that cinobufacini more obviously induced the apoptosis of T-47D cells than that of HBL-100 cells.Western blot analysis confirmed the protein expression of active caspase-3 increased with increasing the dose of cinobufacini.These results indicate that cinobufacini induces the apoptosis of T-47D cells via the up-regulation of caspase-3.
