OBJECTIVE Granulin A(GRN A),a cytokinesis protein,is derived from proteolysis of progranulin.The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly.This study aim...OBJECTIVE Granulin A(GRN A),a cytokinesis protein,is derived from proteolysis of progranulin.The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly.This study aimed to investigate the effect of combination of GRN A and cisplatin on in vitro and in vivo on the growth of hepatocellular carcinoma.METHODS The in vitro and in vivo antitumor effects of combination of GRN A and Cisplatin were evaluated with MTS assay and subcuta.neous transplantation tumor model.Chou-Talalay method was used to calculate the combination index(CI).Colony formation assay and flow cytometry were used to detect the effects of GRN A on apoptosis.The expression of apoptosis-related proteins were detected by Western blot.RESULTS MTS assay showed that GRN A significantly inhibit hepatocellular carcinoma cells growth with the IC50 of 5.6 μmol·L^(-1),and GRN A combined with cisplatin synergistically inhibit hepatocellular carcinoma proliferation,with the CI<1.The colony-formation assay showed that GRN A significantly enhanced the inhibitory effects of cisplatin on cellular anchorage-independent growth.Flow cytometry showed that GRN A combined with cisplatin synergistically induced apoptosis,with the apoptotic rates of 5.87%,32.74%,35.67% and 67.15% in control,GRN A,Cisplatin,and combination of GRN A and Cisplatin groups,respectively.Western blot confirmed that the two drugs synergistically changed the expressions of proteins related to apoptosis.In vivo experiment indicated that combination of GRN A and cisplatin significantly suppressed tumor growth compared with single drug treatment groups.CONCLUSION The combination of GRN A and cisplatin resulted in synergistic antitumor effects against hepatocellular carcinoma both in vitro and in vivo.展开更多
OBJECTIVE Zn-doped CuO nanocomposites(Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells...OBJECTIVE Zn-doped CuO nanocomposites(Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells,as well as the potential mechanisms.METHODS MTS assay was used to detect the effects of Zn-CuO NPS on proliferation pancreatic cancer cells(Panc-mia and Aspc-1).The in vivo antitumor effects of Zn-CuO NPs were detected by xenografts model in nude mice.The effects of Zn-CuO NPS on autophagy were detected bytransmission electron microscopy(TEM) andflow cytometry.Autophagy related proteins were detected by Western blotting.RESULTS Zn-CuO NPS significantly inhibited the proliferation of Panc-mia cells and Aspc-1 cells.In vivo experi.ments showed that Zn-CuO NPS significantly inhibited the tumor growth in nude mice without affecting the body weight of the mice.TEM and flow cytometry showed that Zn-CuO NPS induced autophagy,and significantly increased the number of autophagosome.Western Blot showed that Zn-CuO NPS alterd the expression of autophagy related proteins,such as AMPK,mTORand Beclin-1.Also,AMPK inhibitor could significantly reduce Zn-CuO NPS-induced autophagy pathwayas analyzed byWestern blotting.CONCLUSION The findings suggested that Zn-doped CuO nanocomposites inhibited the in vitro and in vivo growth of pancreatic cancer by inducing autophagy through AMPK/mTOR pathway.展开更多
OBJECTIVE Zn-doped CuO nanocomposites(nZn-CuO NPs) are novel nanoparticles synthesized by sonochemical method.This study aimed to further investigate the antitumor effects and mechanism of nZn-CuO NPs,as well as the e...OBJECTIVE Zn-doped CuO nanocomposites(nZn-CuO NPs) are novel nanoparticles synthesized by sonochemical method.This study aimed to further investigate the antitumor effects and mechanism of nZn-CuO NPs,as well as the exact mechanism of reactive oxygen species(ROS) on nZn-CuO NPs-induced death using N-acetylcysteine(NAC).METHODS The antitumor effects of nZn-CuO NPs were evaluated by MTS assay and orthotopic transplantation tumor model in nude mice.The effects of nZn-CuO NPs with or without NAC on ROS production,DNA damage,apoptosis,mitochondrial damage,autophagy,lysosome impairment,and ER and Golgi stress were determined.Also,western blot was used to detect apoptosis and autophagy related proteins,as well as NF-κB pathway related proteins.RESULTS nZn-CuO NPs significantly inhibit tumor growth both in vitro and in vivo.nZn-CuO NPs were able to cause cytotoxicity,ROS production,DAN damage mitochondrial damage,apoptosis,and autophagy,and NAC can attenuate them.Further studies showed that nZn-CuO NPs induced changes of apoptosis,autophagy and NF-κB pathway related proteins,and NAC can restore them.CONCLUSION Overall,our data demonstrated that nZn-CuO NPs could inhibit tumor growth both in vitro and in vivo by ROS-dependent regulation of apoptosis and autophagy,which might be cross-linked by NF-κB pathways.展开更多
OBJECTIVE Cloves(Syzygium aromaticumL.) have been used as both a spice and a traditional Chinese medicinal herb for thousands of years.However,relatively little is known about its potential anticancer activity and mec...OBJECTIVE Cloves(Syzygium aromaticumL.) have been used as both a spice and a traditional Chinese medicinal herb for thousands of years.However,relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti.tumor effects and mechanisms of water extract of cloves(WEC) against colorectal cancer.METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3 a/b) were determined by western blot.RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50 values of 150 μg·mL-1.The colonyformation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice.Flow cytometry result showed that WEC significantly induced autophagy,and the averaged relative values of fluorescence intensity were206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1 WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I.CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo,which might be related with autophagy induction,and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.展开更多
Discovery of drugs rapidly and effectively is an important aspect for Alzheimer's disease(AD).In this study,a novel high-throughput screening(HTS)method aims at screening the small-molecules with amyloid-β(Aβ)bi...Discovery of drugs rapidly and effectively is an important aspect for Alzheimer's disease(AD).In this study,a novel high-throughput screening(HTS)method aims at screening the small-molecules with amyloid-β(Aβ)binding affinity from natural medicines,based on the combinational use of biolayer interferometry(BLI)and ultra-high-performance liquid chromatography coupled with diode-array detector and quadrupole/time-of-flight tandem mass spectrometry(UHPLC−DAD-Q/TOF-MS/MS)has been firstly developed.Briefly,the components in natural medicines disassociated from biotinylated Aβwere collected to analyze their potential Aβbinding affinity by UHPLC−DAD-Q/TOF-MS/MS.Here,baicalein was confirmed to exhibit the highest binding affinity with Aβin Scutellaria baicalensis.Moreover,polyporenic acid C(PPAC),dehydrotumulosic acid(DTA),and tumulosic acid(TA)in Kai-Xin-San(KXS)were also identified as potent Aβinhibitors.Further bioactivity validations indicated that these compounds could inhibit Aβfibrillation,improve the viability in Aβ-induced PC-12 cells,and decrease the Aβcontent and improve the behavioral ability in Caenorhabditis elegans.The molecular docking results confirmed that PPAC,DTA,and TA possessed good binding properties with Aβ.Collectively,the present study has provided a novel and effective HTS method for the identification of natural inhibitors on Aβfibrillation,which may accelerate the process on anti-AD drugs discovery and development.展开更多
文摘OBJECTIVE Granulin A(GRN A),a cytokinesis protein,is derived from proteolysis of progranulin.The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly.This study aimed to investigate the effect of combination of GRN A and cisplatin on in vitro and in vivo on the growth of hepatocellular carcinoma.METHODS The in vitro and in vivo antitumor effects of combination of GRN A and Cisplatin were evaluated with MTS assay and subcuta.neous transplantation tumor model.Chou-Talalay method was used to calculate the combination index(CI).Colony formation assay and flow cytometry were used to detect the effects of GRN A on apoptosis.The expression of apoptosis-related proteins were detected by Western blot.RESULTS MTS assay showed that GRN A significantly inhibit hepatocellular carcinoma cells growth with the IC50 of 5.6 μmol·L^(-1),and GRN A combined with cisplatin synergistically inhibit hepatocellular carcinoma proliferation,with the CI<1.The colony-formation assay showed that GRN A significantly enhanced the inhibitory effects of cisplatin on cellular anchorage-independent growth.Flow cytometry showed that GRN A combined with cisplatin synergistically induced apoptosis,with the apoptotic rates of 5.87%,32.74%,35.67% and 67.15% in control,GRN A,Cisplatin,and combination of GRN A and Cisplatin groups,respectively.Western blot confirmed that the two drugs synergistically changed the expressions of proteins related to apoptosis.In vivo experiment indicated that combination of GRN A and cisplatin significantly suppressed tumor growth compared with single drug treatment groups.CONCLUSION The combination of GRN A and cisplatin resulted in synergistic antitumor effects against hepatocellular carcinoma both in vitro and in vivo.
文摘OBJECTIVE Zn-doped CuO nanocomposites(Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells,as well as the potential mechanisms.METHODS MTS assay was used to detect the effects of Zn-CuO NPS on proliferation pancreatic cancer cells(Panc-mia and Aspc-1).The in vivo antitumor effects of Zn-CuO NPs were detected by xenografts model in nude mice.The effects of Zn-CuO NPS on autophagy were detected bytransmission electron microscopy(TEM) andflow cytometry.Autophagy related proteins were detected by Western blotting.RESULTS Zn-CuO NPS significantly inhibited the proliferation of Panc-mia cells and Aspc-1 cells.In vivo experi.ments showed that Zn-CuO NPS significantly inhibited the tumor growth in nude mice without affecting the body weight of the mice.TEM and flow cytometry showed that Zn-CuO NPS induced autophagy,and significantly increased the number of autophagosome.Western Blot showed that Zn-CuO NPS alterd the expression of autophagy related proteins,such as AMPK,mTORand Beclin-1.Also,AMPK inhibitor could significantly reduce Zn-CuO NPS-induced autophagy pathwayas analyzed byWestern blotting.CONCLUSION The findings suggested that Zn-doped CuO nanocomposites inhibited the in vitro and in vivo growth of pancreatic cancer by inducing autophagy through AMPK/mTOR pathway.
基金supported by Natural Science Foundation of China(81774191) Beijing Natural Science Foundation(7172031) Project of High-level Teachers in Beijing Municipal Universities in the Period of 13th5-year Plan(CIT＆TCD201804086)
文摘OBJECTIVE Zn-doped CuO nanocomposites(nZn-CuO NPs) are novel nanoparticles synthesized by sonochemical method.This study aimed to further investigate the antitumor effects and mechanism of nZn-CuO NPs,as well as the exact mechanism of reactive oxygen species(ROS) on nZn-CuO NPs-induced death using N-acetylcysteine(NAC).METHODS The antitumor effects of nZn-CuO NPs were evaluated by MTS assay and orthotopic transplantation tumor model in nude mice.The effects of nZn-CuO NPs with or without NAC on ROS production,DNA damage,apoptosis,mitochondrial damage,autophagy,lysosome impairment,and ER and Golgi stress were determined.Also,western blot was used to detect apoptosis and autophagy related proteins,as well as NF-κB pathway related proteins.RESULTS nZn-CuO NPs significantly inhibit tumor growth both in vitro and in vivo.nZn-CuO NPs were able to cause cytotoxicity,ROS production,DAN damage mitochondrial damage,apoptosis,and autophagy,and NAC can attenuate them.Further studies showed that nZn-CuO NPs induced changes of apoptosis,autophagy and NF-κB pathway related proteins,and NAC can restore them.CONCLUSION Overall,our data demonstrated that nZn-CuO NPs could inhibit tumor growth both in vitro and in vivo by ROS-dependent regulation of apoptosis and autophagy,which might be cross-linked by NF-κB pathways.
文摘OBJECTIVE Cloves(Syzygium aromaticumL.) have been used as both a spice and a traditional Chinese medicinal herb for thousands of years.However,relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti.tumor effects and mechanisms of water extract of cloves(WEC) against colorectal cancer.METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3 a/b) were determined by western blot.RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50 values of 150 μg·mL-1.The colonyformation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice.Flow cytometry result showed that WEC significantly induced autophagy,and the averaged relative values of fluorescence intensity were206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1 WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I.CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo,which might be related with autophagy induction,and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.
基金This work was supported by grants from the National Natural Science Foundation of China(81903829,81801398)Department of Science and Technology of Sichuan Province,China(2018JY0474,2019JDPT0010,2021YJ0180,21RCYJ0021,202086,2019YFSY0014)+1 种基金The Joint project of Luzhou Municipal People’s Government and Southwest Medical University,China(2019LZXNYDJ02,2019LZXNYDJ05,2020LZXNYDJ37)The Science and Technology Planning Programs of Luzhou(2018-JYJ-34,China).
文摘Discovery of drugs rapidly and effectively is an important aspect for Alzheimer's disease(AD).In this study,a novel high-throughput screening(HTS)method aims at screening the small-molecules with amyloid-β(Aβ)binding affinity from natural medicines,based on the combinational use of biolayer interferometry(BLI)and ultra-high-performance liquid chromatography coupled with diode-array detector and quadrupole/time-of-flight tandem mass spectrometry(UHPLC−DAD-Q/TOF-MS/MS)has been firstly developed.Briefly,the components in natural medicines disassociated from biotinylated Aβwere collected to analyze their potential Aβbinding affinity by UHPLC−DAD-Q/TOF-MS/MS.Here,baicalein was confirmed to exhibit the highest binding affinity with Aβin Scutellaria baicalensis.Moreover,polyporenic acid C(PPAC),dehydrotumulosic acid(DTA),and tumulosic acid(TA)in Kai-Xin-San(KXS)were also identified as potent Aβinhibitors.Further bioactivity validations indicated that these compounds could inhibit Aβfibrillation,improve the viability in Aβ-induced PC-12 cells,and decrease the Aβcontent and improve the behavioral ability in Caenorhabditis elegans.The molecular docking results confirmed that PPAC,DTA,and TA possessed good binding properties with Aβ.Collectively,the present study has provided a novel and effective HTS method for the identification of natural inhibitors on Aβfibrillation,which may accelerate the process on anti-AD drugs discovery and development.
