Objective:Despite the combination of Scutellaria barbata D.Don and Scleromitrion diffusum(Willd.)R.J.Wang(SB-SD)being a recognized Chinese medicinal herbal pair that is commonly used in the treatment of ovarian cancer...Objective:Despite the combination of Scutellaria barbata D.Don and Scleromitrion diffusum(Willd.)R.J.Wang(SB-SD)being a recognized Chinese medicinal herbal pair that is commonly used in the treatment of ovarian cancer,there is a poor understanding of their pharmacological mechanisms.This study examines the antitumor properties and potential mechanisms of SB-SD on human ovarian cancer A2780 cells through a multi-omics approach,establishing a pharmacological basis for clinical utilization.Methods:A range of mass ratios and reagents were used in the hot reflux extraction of SB-SD.The inhibitory effect of the SB-SD extracts on A2780 cell proliferation was assessed using the cell-counting kit 8assay.A zebrafish tumor implantation model was used to evaluate the effects of SB-SD extracts on tumor growth and metastasis in vivo.Transcriptomics and proteomics were used to investigate alterations in biological pathways in A2780 cells after treatment with different concentrations of SB-SD extract.Cell cycle,cell apoptosis,intracellular free iron concentration,intracellular reactive oxygen species(ROS)concentration,malondialdehyde(MDA),and mitochondrial membrane potential were measured.Real-time quantitative reverse transcription polymerase chain reaction and Western blotting were utilized to investigate the effects of heme catabolism and ferritinophagy on ferroptosis induced by SB-SD extract in A2780 cells.Results:The 70%ethanol extract of SB-SD(a mass ratio of 4:1)inhibited A2780 cell proliferation significantly with a half maximal inhibitory concentration of 660μg/m L in a concentration-and timedependent manner.Moreover,it effectively suppressed tumor growth and metastasis in a zebrafish tumor implantation model.SB-SD extract induced the accumulation of free iron,ROS,MDA,and mitochondrial damage in A2780 cells.The mechanisms might involve the upregulated expression of ferritinophagyrelated genes microtubule-associated protein 1 light chain 3,autophagy-related gene 5,and nuclear receptor coactivator 4.Conclusion:SB-SD extract effectively inhibited the development of ovarian cancer both in vitro and in vivo.Its mechanism of action involved inducing ferroptosis by facilitating heme catabolism and ferritinophagy.This herbal pair holds promise as a potential therapeutic option for ovarian cancer treatment and may be utilized in combination with routine treatment to improve the treatment outcomes of ovarian cancer patients.展开更多
Objective： To investigate the effect of the ethanol extract of Scutellaria barbata D. Don（EESB） on colorectal cancer（CRC） growth and Wnt/β-catenin signaling pathway in vivo and in vitro. Methods： In vivoexperim...Objective： To investigate the effect of the ethanol extract of Scutellaria barbata D. Don（EESB） on colorectal cancer（CRC） growth and Wnt/β-catenin signaling pathway in vivo and in vitro. Methods： In vivoexperiment, CRC xenograft mouse model was constructed with injection of HT-29 cells. Following xenograft implantation, twenty mice were randomly divided into EESB-treated group（n=10） and control group（n=10） by a random number table, and were given with intra-gastric administration of 2 g/kg EESB or saline, 5 days a week for 16 days, respectively. At the end of experiment, tumors were removed and weighed by electronic scales. The proliferation biomarker Ki-67 of tumor was evaluated by immunohistochemistry（IHC） assay. In vitro study, HT-29 cells were treated with 0, 0.5, 1.5, 2.5 mg/m L EESB for 24 h. At the end of the treatment, the viability and survival of HT-29 cells were determined by methylthiazolyldiphenyl-tetrazolium bromide（MTT） assay and colony formation assay, respectively. The m RNA expression of c-Myc, Survivin and adenomatous polyposis coli（APC） was examined by reverse transcription-polymerase chain reaction（RT-PCR） both in tumor tissues of CRC xenograft mice and HT-29 cells. Protein expression of c-Myc, Survivin, APC, and β-catenin as well as β-catenin phosphorylation level were evaluated by IHC assay or Western blotting. Results： EESB significantly reduced tumor weight in CRC xenografts mice, compared with the control group（P〈0.05）. IHC assay showed that EESB significantly inhibited protein expression of Ki-67 in tumor tissues（P〈0.05）. MTT assay showed that EESB significantly reduced HT-29 cell viability in a dose-dependent manner（P〈0.05）. Colony formation assay showed that EESB dose-dependently decreased the survival of HT-29 cells（P〈0.05）. In addition, RT-PCR assay showed that EESB decreased the m RNA expression of c-Myc and Survivin and increased APC expression, both in tumor tissues of CRC xenograft mice and HT-29 cells（P〈0.05）. IHC assay or Western blotting showed that EESB decreased protein expression of β-catenin, c-Myc and Survivin, as well as increased APC expression and β-catenin phosphorylation in tumor tissues or HT-29 cells（P〈0.05）. Conclusions： EESB significantly reduced tumor growth in CRC xenografts mice, and inhibited the viability and survival of HT-29 cells. EESB could suppress the activation of the Wnt/β-catenin pathway, which might be one of the mechanisms whereby Scutellaria barbata D. Don exerts its anticancer activity.展开更多
基金supported by the National Natural Science Foundation of China(No.81873195)the Applied Basic Research Program of Liaoning Province(No.2023JH2/101300103)+1 种基金the Liaoning Revitalization Talents Program(No.XLYC1907113)the Dalian Medical University Foundation for Teaching Reform Project of Undergraduate Innovative Talents Training(No.111807010303)。
文摘Objective:Despite the combination of Scutellaria barbata D.Don and Scleromitrion diffusum(Willd.)R.J.Wang(SB-SD)being a recognized Chinese medicinal herbal pair that is commonly used in the treatment of ovarian cancer,there is a poor understanding of their pharmacological mechanisms.This study examines the antitumor properties and potential mechanisms of SB-SD on human ovarian cancer A2780 cells through a multi-omics approach,establishing a pharmacological basis for clinical utilization.Methods:A range of mass ratios and reagents were used in the hot reflux extraction of SB-SD.The inhibitory effect of the SB-SD extracts on A2780 cell proliferation was assessed using the cell-counting kit 8assay.A zebrafish tumor implantation model was used to evaluate the effects of SB-SD extracts on tumor growth and metastasis in vivo.Transcriptomics and proteomics were used to investigate alterations in biological pathways in A2780 cells after treatment with different concentrations of SB-SD extract.Cell cycle,cell apoptosis,intracellular free iron concentration,intracellular reactive oxygen species(ROS)concentration,malondialdehyde(MDA),and mitochondrial membrane potential were measured.Real-time quantitative reverse transcription polymerase chain reaction and Western blotting were utilized to investigate the effects of heme catabolism and ferritinophagy on ferroptosis induced by SB-SD extract in A2780 cells.Results:The 70%ethanol extract of SB-SD(a mass ratio of 4:1)inhibited A2780 cell proliferation significantly with a half maximal inhibitory concentration of 660μg/m L in a concentration-and timedependent manner.Moreover,it effectively suppressed tumor growth and metastasis in a zebrafish tumor implantation model.SB-SD extract induced the accumulation of free iron,ROS,MDA,and mitochondrial damage in A2780 cells.The mechanisms might involve the upregulated expression of ferritinophagyrelated genes microtubule-associated protein 1 light chain 3,autophagy-related gene 5,and nuclear receptor coactivator 4.Conclusion:SB-SD extract effectively inhibited the development of ovarian cancer both in vitro and in vivo.Its mechanism of action involved inducing ferroptosis by facilitating heme catabolism and ferritinophagy.This herbal pair holds promise as a potential therapeutic option for ovarian cancer treatment and may be utilized in combination with routine treatment to improve the treatment outcomes of ovarian cancer patients.
基金Supported by the Natural Science Foundation of Fujian Province of China(No.2013J01333)the Youth Science Foundation of the Health Department of Fujian Province(2012-2-60)the Developmental Fund of Chen Keji Integrative Medicine(No.CKJ2015008)
文摘Objective： To investigate the effect of the ethanol extract of Scutellaria barbata D. Don（EESB） on colorectal cancer（CRC） growth and Wnt/β-catenin signaling pathway in vivo and in vitro. Methods： In vivoexperiment, CRC xenograft mouse model was constructed with injection of HT-29 cells. Following xenograft implantation, twenty mice were randomly divided into EESB-treated group（n=10） and control group（n=10） by a random number table, and were given with intra-gastric administration of 2 g/kg EESB or saline, 5 days a week for 16 days, respectively. At the end of experiment, tumors were removed and weighed by electronic scales. The proliferation biomarker Ki-67 of tumor was evaluated by immunohistochemistry（IHC） assay. In vitro study, HT-29 cells were treated with 0, 0.5, 1.5, 2.5 mg/m L EESB for 24 h. At the end of the treatment, the viability and survival of HT-29 cells were determined by methylthiazolyldiphenyl-tetrazolium bromide（MTT） assay and colony formation assay, respectively. The m RNA expression of c-Myc, Survivin and adenomatous polyposis coli（APC） was examined by reverse transcription-polymerase chain reaction（RT-PCR） both in tumor tissues of CRC xenograft mice and HT-29 cells. Protein expression of c-Myc, Survivin, APC, and β-catenin as well as β-catenin phosphorylation level were evaluated by IHC assay or Western blotting. Results： EESB significantly reduced tumor weight in CRC xenografts mice, compared with the control group（P〈0.05）. IHC assay showed that EESB significantly inhibited protein expression of Ki-67 in tumor tissues（P〈0.05）. MTT assay showed that EESB significantly reduced HT-29 cell viability in a dose-dependent manner（P〈0.05）. Colony formation assay showed that EESB dose-dependently decreased the survival of HT-29 cells（P〈0.05）. In addition, RT-PCR assay showed that EESB decreased the m RNA expression of c-Myc and Survivin and increased APC expression, both in tumor tissues of CRC xenograft mice and HT-29 cells（P〈0.05）. IHC assay or Western blotting showed that EESB decreased protein expression of β-catenin, c-Myc and Survivin, as well as increased APC expression and β-catenin phosphorylation in tumor tissues or HT-29 cells（P〈0.05）. Conclusions： EESB significantly reduced tumor growth in CRC xenografts mice, and inhibited the viability and survival of HT-29 cells. EESB could suppress the activation of the Wnt/β-catenin pathway, which might be one of the mechanisms whereby Scutellaria barbata D. Don exerts its anticancer activity.
