背景:鼻咽癌(Nasopharyngeal carcinoma, NPC)是一种源于鼻咽上皮的恶性肿瘤。Alisol A是一种来源于泽泻根茎的三萜类化合物,具有抑制癌细胞生长和诱导癌细胞凋亡的能力。Alisol A对鼻咽癌的影响尚不明确。方法:Western blot检测蛋白表...背景:鼻咽癌(Nasopharyngeal carcinoma, NPC)是一种源于鼻咽上皮的恶性肿瘤。Alisol A是一种来源于泽泻根茎的三萜类化合物,具有抑制癌细胞生长和诱导癌细胞凋亡的能力。Alisol A对鼻咽癌的影响尚不明确。方法:Western blot检测蛋白表达。采用AutoDock Vina和Discovery Studio软件进行分子对接。结果:Alisol A可抑制鼻咽癌细胞的活力、增殖、迁移和侵袭。分子对接模拟实验证实Alisol A与YAP蛋白结合。此外,在鼻咽癌细胞中,Alisol A促进YAP的磷酸化并抑制YAP的表达。结论:Alisol A通过抑制Hippo信号通路抑制鼻咽癌细胞。Alisol A可能是治疗鼻咽癌的候选药物。Background: Nasopharyngeal carcinoma (NPC) is a malignant tumor originating from the nasopharyngeal epithelium. Alisol A, a triterpenoid compound derived from rhizome of Alismatis alismatis, has been shown to inhibit cancer cell growth and induce apoptosis. The effect of Alisol A on nasopharyngeal carcinoma (NPC) is still unclear. Methods: Western blot was used to detect protein expression. AutoDock Vina and Discovery Studio software were used for molecular docking. Results: Alisol A inhibited the viability, proliferation, migration, and invasion of NPC cells. The molecular docking simulation assay confirmed that Alisol A is bound to YAP protein. In addition, Alisol A promoted the phosphorylation of YAP and suppressed the expression of YAP in NPC cells. Conclusion: Alisol A inhibits the growth of NPC cells by inhibiting Hippo signaling pathway. Alisol A may be a candidate drug for the treatment of NPC.展开更多
目的:Alisol A是从泽泻中分离得到的具有生物活性的三萜类化合物,具有抗癌潜能。本研究旨在探讨Alisol A对鼻咽癌细胞生长的影响。方法:MTT实验、集落形成实验、流式细胞术、transwell实验、伤口愈合实验分别检测细胞活力、增殖、细胞...目的:Alisol A是从泽泻中分离得到的具有生物活性的三萜类化合物,具有抗癌潜能。本研究旨在探讨Alisol A对鼻咽癌细胞生长的影响。方法:MTT实验、集落形成实验、流式细胞术、transwell实验、伤口愈合实验分别检测细胞活力、增殖、细胞周期、迁移、侵袭。结果:Alisol A可抑制鼻咽癌细胞的活力、增殖、迁移和侵袭。Alisol A对C666-1和HK1细胞的生长有明显的抑制作用,并呈时间和浓度依赖性。Alisol A处理显著降低了鼻咽癌细胞中细胞周期相关基因的蛋白表达。在Alisol A处理的细胞中,鼻咽癌细胞的迁移和侵袭能力降低。Alisol A处理显著降低了鼻咽癌细胞中MMP2和MMP9的蛋白表达。结论:Alisol A抑制鼻咽癌细胞的增殖、迁移和侵袭。Alisol A可能是治疗鼻咽癌的靶点。Purpose: Alisol A is a bioactive triterpenoid isolated from the Rhizoma alismatis, which has anticancer potential. In this study, we explored the effect of Alisol A on the growth of nasopharyngeal carcinoma (NPC) cells. Methods: MTT assay, colony formation assay, flow cytometry, transwell assay, wound healing assay were used to assess cell viability, proliferation, cell cycle, migration, invasion, respectively, in vitro. Results: Alisol A inhibited the viability, proliferation, migration, and invasion of NPC cells. Alisol A significantly inhibited the growth of C666-1 and HK1 cells in a time- and concentration-dependent manner. Alisol A treatment significantly reduced the protein expression of cell cycle-related genes in NPC cells. The migration and invasion abilities of NPC cells were reduced in Alisol A-treated cells. Alisol A treatment significantly reduced the protein expression of MMP2 and MMP9 in NPC cells. Conclusion: Alisol A inhibited the proliferation, migration, and invasion of NPC cells. Alisol A may be a potential therapeutic target for nasopharyngeal.展开更多
A sensitive and reliable ultra fast liquid chromatography tandem mass spectrometry(UFLC-MS/MS)method has been developed and validated for simultaneous quantitation of alisol A and alisol B 23-acetate from Alisma orien...A sensitive and reliable ultra fast liquid chromatography tandem mass spectrometry(UFLC-MS/MS)method has been developed and validated for simultaneous quantitation of alisol A and alisol B 23-acetate from Alisma orientale(Sam.)Juz.in rat plasma using diazepam as an internal standard(IS).The plasma samples were extracted by liquideliquid extraction with methyl tert-butyl ether and separated on a Venusil MP C18 column(100 mm×2.1 mm,3.0 mm)(Venusil,China)using gradient elution with the mobile phase consisting of methanol and 0.1%acetic acid in water at a flow rate of 0.4 ml/min.The two analytes were monitored with positive electrospray ionization by multiple reaction monitoring mode(MRM).The lower limit of quantitation was 5.00 ng/ml for alisol A and 5.00 ng/ml for alisol B 23-acetate.The calibration curves were linear in the range of 5.00 e2500 ng/ml for alisol A and 5e2500 ng/ml for alisol B 23-acetate.The mean extraction recoveries were above 63.8%for alisol A and 68.0%for alisol B 23-acetate from biological matrixes.Both intra-day and inter-day precision and accuracy of analytes were well within acceptance criteria(15%).The validated method was successfully applied to the pharmacokinetic study of alisol A and alisol B 23-acetate in rat plasma after oral administration of alcohol extract of Alismatis Rhizoma.展开更多
Background:Alisol A is a natural compound isolated from Alismatis Rhizoma,known for its diverse pharmacological activities,including anticancer and neuroprotective effects.This study aimed to explore the anticancer ef...Background:Alisol A is a natural compound isolated from Alismatis Rhizoma,known for its diverse pharmacological activities,including anticancer and neuroprotective effects.This study aimed to explore the anticancer effects of Alisol A on oral cancer cells and elucidate its underlying mechanisms.Methods:Cell viability was measured by MTT assay,cell cycle by flow cytometry,and apoptosis by Annexin V/PI staining and caspase activation.Regulation of signaling pathways was analyzed using an apoptosis-related protein array,immunoblotting,and specific kinase inhibitors.Results:Alisol A reduced the viability of oral cancer cell lines,induced sub-G1 phase accumulation,and augmented the number of apoptotic cells.Protein array results indicated that Alisol A enhanced the expression of heme oxygenase-1(HO-1),while suppressing cellular inhibitor of apoptosis protein 1(cIAP1)and X-linked inhibitor of apoptosis protein(XIAP)levels in SCC-9 cells.These changes were further confirmed in both SCC-9 and HSC-3 cells by immunoblotting.In addition,Alisol A triggered the activation of caspase-8,-9,and-3,as well as poly(ADPribose)polymerase(PARP)cleavage in both cell lines.Analysis of signaling pathways showed that mitogen-activated protein kinases(MAPKs)were significantly activated by Alisol A.Notably,inhibition of JNK and p38markedly reduced Alisol A-induced activation of caspase-8,-9,and-3.Conclusions:Our findings demonstrate that Alisol A exerts potent anticancer effects on oral cancer cells by inducing caspase-dependent apoptosis via activation of the JNK and p38 signaling pathways.These results suggest that Alisol A may have therapeutic potential for the treatment of oral cancer.展开更多
Objective Obesity is a global health concern with management strategies encompassing bariatric surgery and anti-obesity drugs;however,concerns regarding complexities and side effects persist,driving research for more ...Objective Obesity is a global health concern with management strategies encompassing bariatric surgery and anti-obesity drugs;however,concerns regarding complexities and side effects persist,driving research for more effective,low-risk strategies.The promotion of white adipose tissue(WAT)browning has emerged as a promising approach.Moreover,alisol B 23-acetate(AB23A)has demonstrated efficacy in addressing metabolic disorders,suggesting its potential as a therapeutic agent in obesity management.Therefore,in this study,we aimed to investigate the therapeutic potential of AB23A for mitigating obesity by regulating metabolic phenotypes and lipid distribution in mice fed a high-fat diet(HFD).Methods An obesity mouse model was established by administration of an HFD.Glucose and insulin metabolism were assessed via glucose and insulin tolerance tests.Adipocyte size was determined using hematoxylin and eosin staining.The expression of browning markers in WAT was evaluated using Western blotting and quantitative real-time polymerase chain reaction.Metabolic cage monitoring involved the assessment of various parameters,including food and water intake,energy metabolism,respiratory exchange rates,and physical activity.Moreover,oil red O staining was used to evaluate intracellular lipid accumulation.A bioinformatic analysis tool for identifying the molecular mechanisms of traditional Chinese medicine was used to examine AB23A targets and associated signaling pathways.Results AB23A administration significantly reduced the weight of obese mice,decreased the mass of inguinal WAT,epididymal WAT,and perirenal adipose tissue,improved glucose and insulin metabolism,and reduced adipocyte size.Moreover,treatment with AB23A promoted the expression of browning markers in WAT,enhanced overall energy metabolism in mice,and had no discernible effect on food intake,water consumption,or physical activity.In 3T3-L1 cells,AB23A inhibited lipid accumulation,and both AB23A and rapamycin inhibited the mammalian target of rapamycin-sterol regulatory element-binding protein-1(mTOR-SREBP1)signaling pathway.Furthermore,3-isobutyl-1-methylxanthine,dexamethasone and insulin,at concentrations of 0.25 mmol/L,0.25μmol/L and 1μg/mL,respectively,induced activation of the mTOR-SREBP1 signaling pathway,which was further strengthened by an mTOR activator MHY1485.Notably,MHY1485 reversed the beneficial effects of AB23A in 3T3-L1 cells.Conclusion AB23A promoted WAT browning by inhibiting the mTOR-SREBP1 signaling pathway,offering a potential strategy to prevent obesity.展开更多
AIM： To examine the effect of alisol B acetate on the growth of human gastric cancer cell line SGC7901 and its possible mechanism of action.METHODS： The cytotoxic effect of alisol B acetate on SGC7901 cells was meas...AIM： To examine the effect of alisol B acetate on the growth of human gastric cancer cell line SGC7901 and its possible mechanism of action.METHODS： The cytotoxic effect of alisol B acetate on SGC7901 cells was measured by 3-（4,5-dimethylthiazol- 2-yl）-2,5-diphenyltetrazolium bromide （MI-I-） assay. Phase-contrast and electron microscopy were used to observe the morphological changes. Cell cycle and mitochondrial transmembrane potential （A^Pm） were determined by flow cytometry. Western blotting was used to detect the expression of apoptosis-regulated gene Bcl-2, Bax, Apaf-1, caspase-3, caspase-9, Akt, P-Akt and phosphatidylinositol 3-kinases （PI3K）.RESULTS： Alisol B acetate inhibited the proliferation of SGC7901 cell line in a time- and dose-dependent manner. PI staining showed that alisol B acetate can change the cell cycle distribution of SGC7901, increase the proportion of cells in G0-G1 phase and decrease the proportion of S phase cells and G2-M phase cells. Alisol B acetate at a concentration of 30 pmol/L induced apoptosis after 24, 48 and 72 h incubation, with occurrence rates of apoptotic cells of 4.36%, 14.42% and 21.16%, respectively. Phase-contrast and electron microscopy revealed that the nuclear fragmentation and chromosomal condensed, cells shrank and attachment loss appeared in the SGC7901 treated with alisol B acetate. Apoptosis of SGC7901 cells was associated with cell cycle arrest, caspase-3 and caspase-9 activation, loss of mitochondrial membrane potential and up-regulation of the ratio of Bax/Bcl-2 and inhibition of the PI3K/Akt.CONCLUSION： Alisol B acetate exhibits an antiproliferative effect in SGC7901 cells by inducing apoptosis. Apoptosis of SGC7901 cells involves mitochondria-caspase and PI3K/Akt dependent pathways.展开更多
Objective: The present study aimed to investigate the molecular events in alisol B 23-acetate(ABA) cytotoxic activity against a liver cancer cell line.Methods: First, we employed a quantitative proteomics approach bas...Objective: The present study aimed to investigate the molecular events in alisol B 23-acetate(ABA) cytotoxic activity against a liver cancer cell line.Methods: First, we employed a quantitative proteomics approach based on stable isotope labeling by amino acids in cell culture(SILAC) to identify the different proteins expressed in HepG2 liver cancer cells upon exposure to ABA. Next, bioinformatics analyses through DAVID and STRING on-line tools were used to predict the pathways involved. Finally, we applied functional validation including cell cycle analysis and Western blotting for apoptosis and mTOR pathway-related proteins to confirm the bioinformatics predictions.Results: We identified 330 different proteins with the SILAC-based quantitative proteomics approach. The bioinformatics analysis and the functional validation revealed that the mTOR pathway, ribosome biogenesis, cell cycle, and apoptosis pathways were differentially regulated by ABA. G1 cell cycle arrest, apoptosis and mTOR inhibition were confirmed.Conclusions: ABA, a potential mTOR inhibitor, induces the disruption of ribosomal biogenesis. It also affects the mTOR-MRP axis to cause G1 cell cycle arrest and finally leads to cancer cell apoptosis.展开更多
文摘背景:鼻咽癌(Nasopharyngeal carcinoma, NPC)是一种源于鼻咽上皮的恶性肿瘤。Alisol A是一种来源于泽泻根茎的三萜类化合物,具有抑制癌细胞生长和诱导癌细胞凋亡的能力。Alisol A对鼻咽癌的影响尚不明确。方法:Western blot检测蛋白表达。采用AutoDock Vina和Discovery Studio软件进行分子对接。结果:Alisol A可抑制鼻咽癌细胞的活力、增殖、迁移和侵袭。分子对接模拟实验证实Alisol A与YAP蛋白结合。此外,在鼻咽癌细胞中,Alisol A促进YAP的磷酸化并抑制YAP的表达。结论:Alisol A通过抑制Hippo信号通路抑制鼻咽癌细胞。Alisol A可能是治疗鼻咽癌的候选药物。Background: Nasopharyngeal carcinoma (NPC) is a malignant tumor originating from the nasopharyngeal epithelium. Alisol A, a triterpenoid compound derived from rhizome of Alismatis alismatis, has been shown to inhibit cancer cell growth and induce apoptosis. The effect of Alisol A on nasopharyngeal carcinoma (NPC) is still unclear. Methods: Western blot was used to detect protein expression. AutoDock Vina and Discovery Studio software were used for molecular docking. Results: Alisol A inhibited the viability, proliferation, migration, and invasion of NPC cells. The molecular docking simulation assay confirmed that Alisol A is bound to YAP protein. In addition, Alisol A promoted the phosphorylation of YAP and suppressed the expression of YAP in NPC cells. Conclusion: Alisol A inhibits the growth of NPC cells by inhibiting Hippo signaling pathway. Alisol A may be a candidate drug for the treatment of NPC.
文摘目的:Alisol A是从泽泻中分离得到的具有生物活性的三萜类化合物,具有抗癌潜能。本研究旨在探讨Alisol A对鼻咽癌细胞生长的影响。方法:MTT实验、集落形成实验、流式细胞术、transwell实验、伤口愈合实验分别检测细胞活力、增殖、细胞周期、迁移、侵袭。结果:Alisol A可抑制鼻咽癌细胞的活力、增殖、迁移和侵袭。Alisol A对C666-1和HK1细胞的生长有明显的抑制作用,并呈时间和浓度依赖性。Alisol A处理显著降低了鼻咽癌细胞中细胞周期相关基因的蛋白表达。在Alisol A处理的细胞中,鼻咽癌细胞的迁移和侵袭能力降低。Alisol A处理显著降低了鼻咽癌细胞中MMP2和MMP9的蛋白表达。结论:Alisol A抑制鼻咽癌细胞的增殖、迁移和侵袭。Alisol A可能是治疗鼻咽癌的靶点。Purpose: Alisol A is a bioactive triterpenoid isolated from the Rhizoma alismatis, which has anticancer potential. In this study, we explored the effect of Alisol A on the growth of nasopharyngeal carcinoma (NPC) cells. Methods: MTT assay, colony formation assay, flow cytometry, transwell assay, wound healing assay were used to assess cell viability, proliferation, cell cycle, migration, invasion, respectively, in vitro. Results: Alisol A inhibited the viability, proliferation, migration, and invasion of NPC cells. Alisol A significantly inhibited the growth of C666-1 and HK1 cells in a time- and concentration-dependent manner. Alisol A treatment significantly reduced the protein expression of cell cycle-related genes in NPC cells. The migration and invasion abilities of NPC cells were reduced in Alisol A-treated cells. Alisol A treatment significantly reduced the protein expression of MMP2 and MMP9 in NPC cells. Conclusion: Alisol A inhibited the proliferation, migration, and invasion of NPC cells. Alisol A may be a potential therapeutic target for nasopharyngeal.
基金This study was financially supported by Liaoning Innovative Research Team in University(LNIRT,Grant No.LT2013022).
文摘A sensitive and reliable ultra fast liquid chromatography tandem mass spectrometry(UFLC-MS/MS)method has been developed and validated for simultaneous quantitation of alisol A and alisol B 23-acetate from Alisma orientale(Sam.)Juz.in rat plasma using diazepam as an internal standard(IS).The plasma samples were extracted by liquideliquid extraction with methyl tert-butyl ether and separated on a Venusil MP C18 column(100 mm×2.1 mm,3.0 mm)(Venusil,China)using gradient elution with the mobile phase consisting of methanol and 0.1%acetic acid in water at a flow rate of 0.4 ml/min.The two analytes were monitored with positive electrospray ionization by multiple reaction monitoring mode(MRM).The lower limit of quantitation was 5.00 ng/ml for alisol A and 5.00 ng/ml for alisol B 23-acetate.The calibration curves were linear in the range of 5.00 e2500 ng/ml for alisol A and 5e2500 ng/ml for alisol B 23-acetate.The mean extraction recoveries were above 63.8%for alisol A and 68.0%for alisol B 23-acetate from biological matrixes.Both intra-day and inter-day precision and accuracy of analytes were well within acceptance criteria(15%).The validated method was successfully applied to the pharmacokinetic study of alisol A and alisol B 23-acetate in rat plasma after oral administration of alcohol extract of Alismatis Rhizoma.
基金supported by Chung Shan Medical University Hospital,Taiwan(CSH-2024-C-060).
文摘Background:Alisol A is a natural compound isolated from Alismatis Rhizoma,known for its diverse pharmacological activities,including anticancer and neuroprotective effects.This study aimed to explore the anticancer effects of Alisol A on oral cancer cells and elucidate its underlying mechanisms.Methods:Cell viability was measured by MTT assay,cell cycle by flow cytometry,and apoptosis by Annexin V/PI staining and caspase activation.Regulation of signaling pathways was analyzed using an apoptosis-related protein array,immunoblotting,and specific kinase inhibitors.Results:Alisol A reduced the viability of oral cancer cell lines,induced sub-G1 phase accumulation,and augmented the number of apoptotic cells.Protein array results indicated that Alisol A enhanced the expression of heme oxygenase-1(HO-1),while suppressing cellular inhibitor of apoptosis protein 1(cIAP1)and X-linked inhibitor of apoptosis protein(XIAP)levels in SCC-9 cells.These changes were further confirmed in both SCC-9 and HSC-3 cells by immunoblotting.In addition,Alisol A triggered the activation of caspase-8,-9,and-3,as well as poly(ADPribose)polymerase(PARP)cleavage in both cell lines.Analysis of signaling pathways showed that mitogen-activated protein kinases(MAPKs)were significantly activated by Alisol A.Notably,inhibition of JNK and p38markedly reduced Alisol A-induced activation of caspase-8,-9,and-3.Conclusions:Our findings demonstrate that Alisol A exerts potent anticancer effects on oral cancer cells by inducing caspase-dependent apoptosis via activation of the JNK and p38 signaling pathways.These results suggest that Alisol A may have therapeutic potential for the treatment of oral cancer.
基金supported by Shandong Provincial Natural Science Foundation General Program(No.ZR2022MH213)Shandong Provincial Traditional Chinese Medicine Science and Technology Project General Program(No.M2023241)+1 种基金Jinan Clinical Medical Science and Technology Innovation Program(No.202328013)Qinghai Province High-end Innovative Talents Thousand Talents Program.
文摘Objective Obesity is a global health concern with management strategies encompassing bariatric surgery and anti-obesity drugs;however,concerns regarding complexities and side effects persist,driving research for more effective,low-risk strategies.The promotion of white adipose tissue(WAT)browning has emerged as a promising approach.Moreover,alisol B 23-acetate(AB23A)has demonstrated efficacy in addressing metabolic disorders,suggesting its potential as a therapeutic agent in obesity management.Therefore,in this study,we aimed to investigate the therapeutic potential of AB23A for mitigating obesity by regulating metabolic phenotypes and lipid distribution in mice fed a high-fat diet(HFD).Methods An obesity mouse model was established by administration of an HFD.Glucose and insulin metabolism were assessed via glucose and insulin tolerance tests.Adipocyte size was determined using hematoxylin and eosin staining.The expression of browning markers in WAT was evaluated using Western blotting and quantitative real-time polymerase chain reaction.Metabolic cage monitoring involved the assessment of various parameters,including food and water intake,energy metabolism,respiratory exchange rates,and physical activity.Moreover,oil red O staining was used to evaluate intracellular lipid accumulation.A bioinformatic analysis tool for identifying the molecular mechanisms of traditional Chinese medicine was used to examine AB23A targets and associated signaling pathways.Results AB23A administration significantly reduced the weight of obese mice,decreased the mass of inguinal WAT,epididymal WAT,and perirenal adipose tissue,improved glucose and insulin metabolism,and reduced adipocyte size.Moreover,treatment with AB23A promoted the expression of browning markers in WAT,enhanced overall energy metabolism in mice,and had no discernible effect on food intake,water consumption,or physical activity.In 3T3-L1 cells,AB23A inhibited lipid accumulation,and both AB23A and rapamycin inhibited the mammalian target of rapamycin-sterol regulatory element-binding protein-1(mTOR-SREBP1)signaling pathway.Furthermore,3-isobutyl-1-methylxanthine,dexamethasone and insulin,at concentrations of 0.25 mmol/L,0.25μmol/L and 1μg/mL,respectively,induced activation of the mTOR-SREBP1 signaling pathway,which was further strengthened by an mTOR activator MHY1485.Notably,MHY1485 reversed the beneficial effects of AB23A in 3T3-L1 cells.Conclusion AB23A promoted WAT browning by inhibiting the mTOR-SREBP1 signaling pathway,offering a potential strategy to prevent obesity.
文摘AIM： To examine the effect of alisol B acetate on the growth of human gastric cancer cell line SGC7901 and its possible mechanism of action.METHODS： The cytotoxic effect of alisol B acetate on SGC7901 cells was measured by 3-（4,5-dimethylthiazol- 2-yl）-2,5-diphenyltetrazolium bromide （MI-I-） assay. Phase-contrast and electron microscopy were used to observe the morphological changes. Cell cycle and mitochondrial transmembrane potential （A^Pm） were determined by flow cytometry. Western blotting was used to detect the expression of apoptosis-regulated gene Bcl-2, Bax, Apaf-1, caspase-3, caspase-9, Akt, P-Akt and phosphatidylinositol 3-kinases （PI3K）.RESULTS： Alisol B acetate inhibited the proliferation of SGC7901 cell line in a time- and dose-dependent manner. PI staining showed that alisol B acetate can change the cell cycle distribution of SGC7901, increase the proportion of cells in G0-G1 phase and decrease the proportion of S phase cells and G2-M phase cells. Alisol B acetate at a concentration of 30 pmol/L induced apoptosis after 24, 48 and 72 h incubation, with occurrence rates of apoptotic cells of 4.36%, 14.42% and 21.16%, respectively. Phase-contrast and electron microscopy revealed that the nuclear fragmentation and chromosomal condensed, cells shrank and attachment loss appeared in the SGC7901 treated with alisol B acetate. Apoptosis of SGC7901 cells was associated with cell cycle arrest, caspase-3 and caspase-9 activation, loss of mitochondrial membrane potential and up-regulation of the ratio of Bax/Bcl-2 and inhibition of the PI3K/Akt.CONCLUSION： Alisol B acetate exhibits an antiproliferative effect in SGC7901 cells by inducing apoptosis. Apoptosis of SGC7901 cells involves mitochondria-caspase and PI3K/Akt dependent pathways.
基金supported by the National Natural Science Foundation of China (No. NSFC-81502406, 81673320)Education Scientific Research Project for young teachers, 2018 (No. JT180013)
文摘Objective: The present study aimed to investigate the molecular events in alisol B 23-acetate(ABA) cytotoxic activity against a liver cancer cell line.Methods: First, we employed a quantitative proteomics approach based on stable isotope labeling by amino acids in cell culture(SILAC) to identify the different proteins expressed in HepG2 liver cancer cells upon exposure to ABA. Next, bioinformatics analyses through DAVID and STRING on-line tools were used to predict the pathways involved. Finally, we applied functional validation including cell cycle analysis and Western blotting for apoptosis and mTOR pathway-related proteins to confirm the bioinformatics predictions.Results: We identified 330 different proteins with the SILAC-based quantitative proteomics approach. The bioinformatics analysis and the functional validation revealed that the mTOR pathway, ribosome biogenesis, cell cycle, and apoptosis pathways were differentially regulated by ABA. G1 cell cycle arrest, apoptosis and mTOR inhibition were confirmed.Conclusions: ABA, a potential mTOR inhibitor, induces the disruption of ribosomal biogenesis. It also affects the mTOR-MRP axis to cause G1 cell cycle arrest and finally leads to cancer cell apoptosis.
