The aim of this study was to investigate the effect of Paris saponin I （PSI ） on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms. The proliferation of SGC7901 cells was moni...The aim of this study was to investigate the effect of Paris saponin I （PSI ） on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms. The proliferation of SGC7901 cells was monitored by the MTT cell viability assay, while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining. Flow cytometry was performed to analyze the cell cycle progression of propidium iodide （PI）-stained SGC7901 cells and the apoptotic rate of annexin V/PI-stained ceils. Western blotting was used to examine the expression of several cell cycle proteins, including cyclin 131 and Cdkl, and the apoptosis-regulated proteins Bcl-2, Bax, cytochrome c, procaspase-9, and procaspase-3. The MTT assay demonstrated that PSI could induce significant dose- and time-dependent inhibition of SGC7901 cell proliferation. Marked morphological changes, including condensation of chromatin, nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining. PS I treatment also resulted in the disruption of the cell cycle at Gz/M and the induction of apoptosis. Following PSI treatment, the cell cycle-related proteins cyclin B 1 and Cdkl were down- regulated. Expression of the pro-apoptotic protein Bax was increased, while anti-apoptotic protein Bcl-2 decreased. PSI treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3. These data indicate that PS I acts as an inhibitor of proliferation in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis. PSI is a potential therapeutic agent against human gastric carcinoma.展开更多
Paris saponinⅦ(PSⅦ),a bioactive constituent extracted from Trillium tschonoskii Maxim.,is cytotoxic to several cancer types.This study was designed to explore whether PSⅦprevents non-small-cell lung cancer(NSCLC)pr...Paris saponinⅦ(PSⅦ),a bioactive constituent extracted from Trillium tschonoskii Maxim.,is cytotoxic to several cancer types.This study was designed to explore whether PSⅦprevents non-small-cell lung cancer(NSCLC)proliferation and to investigate its molecular target.AMP-activated protein kinase(AMPK)has been implicated in the activation of autophagy in distinct tissues.In cultured human NSCLC cell lines,PSⅦinduces autophagy by activating AMPK and inhibiting m TOR signaling.Furthermore,PSⅦ-induced autophagy activation was reversed by the AMPK inhibitor compound C.Computational docking analysis showed that PSⅦdirectly interacted with the allosteric drug and metabolite site of AMPK to stabilize its activation.Microscale thermophoresis assay and drug affinity responsive target stability assay further confirmed the high affinity between PSⅦand AMPK.In summary,PSⅦacts as a direct AMPK activator to induce cell autophagy,which inhibits the growth of NSCLC cells.In the future,PSⅦtherapy should be applied to treat patients with NSCLC.展开更多
To study the biopharmaceutics characteristics of paris saponin Ⅶ(PSⅦ). The solubility of PSⅦ was evaluated by measurement of the equilibrium solubility in different solvents and media. The permeability of PSⅦ was ...To study the biopharmaceutics characteristics of paris saponin Ⅶ(PSⅦ). The solubility of PSⅦ was evaluated by measurement of the equilibrium solubility in different solvents and media. The permeability of PSⅦ was evaluated by measuring the oil/water partition coefficient(lgP_(app)) and determining the apparent permeability coefficient(PC_(app)) on a mono-layer Caco-2 cell model. The effects of p-glycoprotein and multidrug resistance related protein 2 on PSⅦ transport in mono-layer Caco-2 cell model were further investigated. Finally, the small intestinal absorption of PSⅦ was investigated in rat. In solvents of different pH, the equilibrium solubility of PSⅦ was quite low, and the dose number of PSⅦ was larger than 1. The lgP_(app) of PSⅦ was less than 0. The apparent permeability coefficient [PC_(app)(AP-BL)] of PSⅦ in mono-layer Caco-2 cell model was less than 14.96 × 10^(-6) cm·s^(-1), and the efflux ratio of PSⅦ in mono-layer Caco-2 cell model was less than 1. The transport rate of PSⅦ in mono-layer Caco-2 cell model was not affected by the inhibitors of p-glycoprotein and multidrug resistance related protein 2. After oral administration, PSⅦ could be detected in rat intestinal contents, but could not be detected in the small intestinal mucosa. PSⅦ showed low solubility and permeability,which would result in low oral bioavailability in clinic. PSⅦ belonged to Class IV compound in biopharmaceutics classification system.展开更多
文摘The aim of this study was to investigate the effect of Paris saponin I （PSI ） on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms. The proliferation of SGC7901 cells was monitored by the MTT cell viability assay, while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining. Flow cytometry was performed to analyze the cell cycle progression of propidium iodide （PI）-stained SGC7901 cells and the apoptotic rate of annexin V/PI-stained ceils. Western blotting was used to examine the expression of several cell cycle proteins, including cyclin 131 and Cdkl, and the apoptosis-regulated proteins Bcl-2, Bax, cytochrome c, procaspase-9, and procaspase-3. The MTT assay demonstrated that PSI could induce significant dose- and time-dependent inhibition of SGC7901 cell proliferation. Marked morphological changes, including condensation of chromatin, nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining. PS I treatment also resulted in the disruption of the cell cycle at Gz/M and the induction of apoptosis. Following PSI treatment, the cell cycle-related proteins cyclin B 1 and Cdkl were down- regulated. Expression of the pro-apoptotic protein Bax was increased, while anti-apoptotic protein Bcl-2 decreased. PSI treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3. These data indicate that PS I acts as an inhibitor of proliferation in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis. PSI is a potential therapeutic agent against human gastric carcinoma.
基金supported by the National Natural Science Foundation of China(Nos.82072928 and 81802387)the Foundation for Innovative Research Team of Hubei Provincial Department of Education(No.T201915)+4 种基金Hubei Provincial Technology Innovation Project(No.2017ACA176)the Innovative Research Program for Graduates(No.YC2019001)the Principal Investigator Grant of Hubei University of Medicine(No.HBMUPI201806)the Grants of Open Ended Design Project from Hubei Key Laboratory of Wudang Local Chinese Medicine Research(No.WDCM2019008)the Faculty Development Grants from Hubei University of Medicine(Nos.2018QDJZR03 and 2019QDJZR16)。
文摘Paris saponinⅦ(PSⅦ),a bioactive constituent extracted from Trillium tschonoskii Maxim.,is cytotoxic to several cancer types.This study was designed to explore whether PSⅦprevents non-small-cell lung cancer(NSCLC)proliferation and to investigate its molecular target.AMP-activated protein kinase(AMPK)has been implicated in the activation of autophagy in distinct tissues.In cultured human NSCLC cell lines,PSⅦinduces autophagy by activating AMPK and inhibiting m TOR signaling.Furthermore,PSⅦ-induced autophagy activation was reversed by the AMPK inhibitor compound C.Computational docking analysis showed that PSⅦdirectly interacted with the allosteric drug and metabolite site of AMPK to stabilize its activation.Microscale thermophoresis assay and drug affinity responsive target stability assay further confirmed the high affinity between PSⅦand AMPK.In summary,PSⅦacts as a direct AMPK activator to induce cell autophagy,which inhibits the growth of NSCLC cells.In the future,PSⅦtherapy should be applied to treat patients with NSCLC.
文摘To study the biopharmaceutics characteristics of paris saponin Ⅶ(PSⅦ). The solubility of PSⅦ was evaluated by measurement of the equilibrium solubility in different solvents and media. The permeability of PSⅦ was evaluated by measuring the oil/water partition coefficient(lgP_(app)) and determining the apparent permeability coefficient(PC_(app)) on a mono-layer Caco-2 cell model. The effects of p-glycoprotein and multidrug resistance related protein 2 on PSⅦ transport in mono-layer Caco-2 cell model were further investigated. Finally, the small intestinal absorption of PSⅦ was investigated in rat. In solvents of different pH, the equilibrium solubility of PSⅦ was quite low, and the dose number of PSⅦ was larger than 1. The lgP_(app) of PSⅦ was less than 0. The apparent permeability coefficient [PC_(app)(AP-BL)] of PSⅦ in mono-layer Caco-2 cell model was less than 14.96 × 10^(-6) cm·s^(-1), and the efflux ratio of PSⅦ in mono-layer Caco-2 cell model was less than 1. The transport rate of PSⅦ in mono-layer Caco-2 cell model was not affected by the inhibitors of p-glycoprotein and multidrug resistance related protein 2. After oral administration, PSⅦ could be detected in rat intestinal contents, but could not be detected in the small intestinal mucosa. PSⅦ showed low solubility and permeability,which would result in low oral bioavailability in clinic. PSⅦ belonged to Class IV compound in biopharmaceutics classification system.
