OBJECTIVE Identification of novel autophagy inhibitors for the combinational treatment of non-small cell lung cancer(NSCLC).METHODS MTT assay and annexin V/PI staining assay were used to evaluate the cell proliferatio...OBJECTIVE Identification of novel autophagy inhibitors for the combinational treatment of non-small cell lung cancer(NSCLC).METHODS MTT assay and annexin V/PI staining assay were used to evaluate the cell proliferation and apoptosis,respectively.Immunofluorescence staining and cathepsin activity assay were used to detect autophagy.Small interfering RNA was performed to silence the genes and Western blot assay was used to evaluate the protein express levels.Xenograft experiments were applied for in vivo evaluation.RESULTS Cepharanthine,a natural compound,increased LC3-II expression and GFP-LC3 puncta formation in NSCLC NCI-H1975 cells.Numerous yellow puncta were observed in cepharanthine-treated cells with m RFP-EGFP-LC3 transfection.Co-staining of GFP-LC3 with LysoT racker red or LAMP1 antibody suggested that cepharanthine inhibits autophagosomes-lysosomes fusion.Moreover,cepharanthine attenuated the lysosomal cathepsins maturation.We also confirmed that dacomitinib induced cytoprotective autophagy.Combined treatment with cepharanthine increased the anti-cancer effects of dacomitinib in vitro and in vivo.Besides,cepharanthine could not enhance the anti-cancer effect of dacomitinib in autophagy deficient cells.CONCLUSION Cepharanthine might be further developed as a promising autophagic inhibitor,and combined treatment cepharanthine with dacomitinib could pose as an effective strategy for NSCLC treatment.展开更多
Severe acute respiratory syndrome(SARS) is the first severe viral epidemic we encountered his century,which once spread in more than thirty countriesin2003.1 The etiological agent of SARS has beenc onfirmed to be a n...Severe acute respiratory syndrome(SARS) is the first severe viral epidemic we encountered his century,which once spread in more than thirty countriesin2003.1 The etiological agent of SARS has beenc onfirmed to be a novel coronavirus,namely SARS coronavirus(SARS-CoV),2,3 and the first outbreak of SARS has been successfully controlled world wide,but the identification of SARS-CoV isolated from wildanimals,the emergence of some sporadic SARS cases laterafter that outbreak,all suggest that the recurrence of such an epidemic is not unlikely in the future.In this case,development of SARS vaccines and specific drugs is undoubtedlyessential to the control and prevention from the possible outbreak.4,5展开更多
The Coronavirus disease 2019(COVID-19)has posed a serious threat to global health and the world economy.Antiviral therapies targeting coronavirus are urgently required.The Cepharanthine(CEP)is a traditional Chinese he...The Coronavirus disease 2019(COVID-19)has posed a serious threat to global health and the world economy.Antiviral therapies targeting coronavirus are urgently required.The Cepharanthine(CEP)is a traditional Chinese herbal extract.Our previous research revealed that CEP has a very potent anti-coronavirus effect,but its mechanism of action was not fully understood.To investigate the effect of novel coronavirus on protein glycosylation in infected cells and to further investigate the mechanism of action of CEP against coronavirus,a cellular model using coronavirus GX_P2V infection of Vero E6 cells was established.The effect of coronavirus GX_P2V on host cell protein glycosylation was investigated by N-glycoproteomic analysis,and the antagonistic effect of CEP on the abnormal protein glycosylation caused by coronavirus was analyzed.The results showed that GX_P2V could cause abnormal changes in protein glycosylation levels in host cells,while CEP could partially antagonize the abnormal protein glycosylation caused by GX_P2V.In addition,we also found that CEP could regulate the glycosylation level of coronavirus S protein.In conclusion,this article provides important ideas about the infection mechanism of novel coronaviruses,providing evidence for CEP as a promising therapeutic option for coronavirus infection.展开更多
基金supported by Science and Technology Development Fund,Macao S.A.R(FDCT)(024/2016/A1)Research Fund of University of Macao(MYRG2015-00091-ICMS-QRCM and MYRG2015-00101-ICMS-QRCM)
文摘OBJECTIVE Identification of novel autophagy inhibitors for the combinational treatment of non-small cell lung cancer(NSCLC).METHODS MTT assay and annexin V/PI staining assay were used to evaluate the cell proliferation and apoptosis,respectively.Immunofluorescence staining and cathepsin activity assay were used to detect autophagy.Small interfering RNA was performed to silence the genes and Western blot assay was used to evaluate the protein express levels.Xenograft experiments were applied for in vivo evaluation.RESULTS Cepharanthine,a natural compound,increased LC3-II expression and GFP-LC3 puncta formation in NSCLC NCI-H1975 cells.Numerous yellow puncta were observed in cepharanthine-treated cells with m RFP-EGFP-LC3 transfection.Co-staining of GFP-LC3 with LysoT racker red or LAMP1 antibody suggested that cepharanthine inhibits autophagosomes-lysosomes fusion.Moreover,cepharanthine attenuated the lysosomal cathepsins maturation.We also confirmed that dacomitinib induced cytoprotective autophagy.Combined treatment with cepharanthine increased the anti-cancer effects of dacomitinib in vitro and in vivo.Besides,cepharanthine could not enhance the anti-cancer effect of dacomitinib in autophagy deficient cells.CONCLUSION Cepharanthine might be further developed as a promising autophagic inhibitor,and combined treatment cepharanthine with dacomitinib could pose as an effective strategy for NSCLC treatment.
文摘Severe acute respiratory syndrome(SARS) is the first severe viral epidemic we encountered his century,which once spread in more than thirty countriesin2003.1 The etiological agent of SARS has beenc onfirmed to be a novel coronavirus,namely SARS coronavirus(SARS-CoV),2,3 and the first outbreak of SARS has been successfully controlled world wide,but the identification of SARS-CoV isolated from wildanimals,the emergence of some sporadic SARS cases laterafter that outbreak,all suggest that the recurrence of such an epidemic is not unlikely in the future.In this case,development of SARS vaccines and specific drugs is undoubtedlyessential to the control and prevention from the possible outbreak.4,5
基金the National Key Research and Development Program of China(2018YFA0903000)the National Natural Science Foundation of China(81672001)+2 种基金China MoST Emergency Project on COVID-19(2020YFC0840800)Guangdong Basic and Applied Basic Research Foundation(2021A1515110619)the Fundamental Research Funds for the Central Universities(KG16052901).
文摘The Coronavirus disease 2019(COVID-19)has posed a serious threat to global health and the world economy.Antiviral therapies targeting coronavirus are urgently required.The Cepharanthine(CEP)is a traditional Chinese herbal extract.Our previous research revealed that CEP has a very potent anti-coronavirus effect,but its mechanism of action was not fully understood.To investigate the effect of novel coronavirus on protein glycosylation in infected cells and to further investigate the mechanism of action of CEP against coronavirus,a cellular model using coronavirus GX_P2V infection of Vero E6 cells was established.The effect of coronavirus GX_P2V on host cell protein glycosylation was investigated by N-glycoproteomic analysis,and the antagonistic effect of CEP on the abnormal protein glycosylation caused by coronavirus was analyzed.The results showed that GX_P2V could cause abnormal changes in protein glycosylation levels in host cells,while CEP could partially antagonize the abnormal protein glycosylation caused by GX_P2V.In addition,we also found that CEP could regulate the glycosylation level of coronavirus S protein.In conclusion,this article provides important ideas about the infection mechanism of novel coronaviruses,providing evidence for CEP as a promising therapeutic option for coronavirus infection.
