Objective:To explore the relationship between cancer awareness and the survival of the patients with non-small cell lung carcinoma(NSCLC).Methods:A total of 865 NSCLC patients were screened for the risk factors,includ...Objective:To explore the relationship between cancer awareness and the survival of the patients with non-small cell lung carcinoma(NSCLC).Methods:A total of 865 NSCLC patients were screened for the risk factors,including age,gender,address,tumor/lymph nodes/metastasis(TNM)stage,and cancer awareness.Survival of the patients was calculated by Kaplan-Meier method and Cox regression analysis.Results:After an average observation time of 304 d(ranging from 0 to 4718 d),62 of the 394 patients in the cancer awareness group survived,whereas 26 of the 471 patients in the cancer concealment group survived.Cancer-specific and all-cause survival was poorer in the cancer concealment group(P<0.001 for each,log-rank test).Cox multivariate regression analysis showed that cancer concealment displayed significantly lower cancer-specific survival[hazard ratio(HR)=1.534,95%con fi dence interval(CI)1.320 to 1.784,P<0.001]and all-cause survival(HR=1.558,95%CI 1.346 to 1.803,P<0.001).Conclusion:Cancer concealment is associated with a poor survival of NSCLC patients,which may prohibit the patients from obtaining the real“right to survival”.展开更多
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
OBJECTIVE Lung cancer is the leading cause of cancer death worldwide.Epidermal growth factor receptor(EGFR)mutation(s)is/are common in non-small cell lung cancer(NSCLC)in Asian population,resulting in lung tumor forma...OBJECTIVE Lung cancer is the leading cause of cancer death worldwide.Epidermal growth factor receptor(EGFR)mutation(s)is/are common in non-small cell lung cancer(NSCLC)in Asian population,resulting in lung tumor formation.L858 Rsubstitution mutation on exon 21 and in-flame deletion mutation on exon 19 are the two most common forms of EGFR mutation.Molecular targeted therapy using tyrosine kinase inhibitor(TKI)targeting EGFR shows promising initial response,however drug resistance is common.Therefore,it is needed to identify new inhibitors to tackle TKI-resistance.In this study,we aim to investigate the effect of multiple single purified compounds derived from Chinese herbal medicines(CMHs)on a panel of NSCLC cell lines with different EGFR mutational statuses and TKI sensitivity.We also examine the biological functional effect and drug action mechanism of these cell lines after drug treatment.METHODS We have reviewed the literature and selected ten single purified compounds derived from CMHs which exhibited the highest potential of cancer suppression effect in NSCLC.We have recruited three EGFR-dependent NSCLC cell lines for drug screening using cytotoxicity assay.A549 is used as EGFR wild-type control.Two TKI-resistant NSCLC cell lines were used,H1975 harbors double mutation(EGFRL858R+T790M)and H1650 harbors EGFRexon 19 deletion.H2228is a NSCLC cell line which harbours EML4-ALK fusion gene and was used as EGFR-independent cell line control.MTT assay was used to determine the drug efficacy and IC50 value.Then functional assays including cell cycle arrest analysis and apoptosis assay was used to determine the biological effect after drug treatment.RESULTS MTT assay revealed that six out of ten candidate agents showed significant cancer-inhibiting effects in H1650 and H1975cells.Three compounds exhibited IC50 value at micro-molar levels while another three compounds exhibited IC50 at as low as nano-molar levels.One compound exhibited specificity on EGFR-dependent NSCLC cell lines,which showed 10-fold more selective than the EGFR-independent H2228 cells.Cell cycle analysis and immunoblotting assay showed that one effective compound,designated was MUST-1,altered the metabolic pathway of glucose metabolism and lipid metabolism,and induced cell cycle arrest at G1 phase in NSCLC with EGFR mutation.However,the anti-proliferative effects were distinct in NSCLC cell lines with different EGFR mutation patterns.CONCLUSION MUST-1 significantly induced cell cycle arrest in four NSCLC in EGFR mutant cell lines but the inhibiting effect was not significant in EGFR wildtype cell line.Immunobloting assay revealed that overall intracellular lipid content,glycolytic enzymes PKM2 and cell cycle regulatory gene expression were altered after 72 hcompound treatment in the responsive cells.Further investigation is required to elucidate the underlying reason of drug selectivity and the role of glucose and lipid metabolism in regulating drug sensitivity.展开更多
OBJECTIVE To investigate the pharmacological effect and mechanism of Sanguisorba officinalis L.(SOL)in non-small cell lung cancer(NSCLC)in vitro and in vivo based on network pharmacology.METHODS Network pharmacology w...OBJECTIVE To investigate the pharmacological effect and mechanism of Sanguisorba officinalis L.(SOL)in non-small cell lung cancer(NSCLC)in vitro and in vivo based on network pharmacology.METHODS Network pharmacology was used to analyze the improving effect of SOL on NSCLC and possible targets.Cell counting kit 8(CCK-8)and 5-ethynyl-2′-deoxyuridine(EdU)staining,Western blotting,flow cytometry of AnnexinⅤ/PI,Hoechst 33342/PI staining detection and immunofluorescence were utilized in vitro.H&E staining,immunohistochemistry staining and Western blotting were performed in vivo.RESULTS Based on network prediction,we analyzed the 208 common targets of SOL and NSCLC.36 core targets in 208 common targets were obtained through cytoscape analysis.And the top 10 core targets included Akt,mTOR,EGFR,etc..KEGG analysis showed that PI3K-Akt signaling pathway was the most likely pathway.Furthermore,the mechanism study found that SOL could activate the PI3K/Akt/mTOR signaling pathway in vitro and in vivo.The anti-proliferative effect of SOL in A549 and H1299 cells was measured and validated by CCK-8 and EdU assay.Immunohistochemical results of Ki67 showed that SOL effectively inhibited tumor growth in vivo.SOL also significantly inhibited the migration and invasion of A549 and H1299 cells.SOL significantly increased the percentage of cells with PI signal in A549 and H1299,and the process of cell death of A549 cells indicated that SOL induced apoptosis.The PARP-1 and caspase-3 in A549 and H1299 were found to be activated in a dose manner.The results in vivo were consistent with those in vitro.CONCLUSION SOL-induced,caspase-3-mediated apoptosis via the induction of the PI3K/Akt/mTOR signaling pathway in NSCLC,which further clarified the mechanism of SOL in the inhibition of NSCLC,and thereby provided a possibility for SOL to serve as a novel therapeutic agent for NSCLC in the future.展开更多
OBJECTIVE To explored the potential of pharmacological stabilization and reactivation of p53 for targeted cancer therapies.METHODS The cytotoxicity of a potent Cyclophilin A(CypA)inhibitor HL001 was tasted against a p...OBJECTIVE To explored the potential of pharmacological stabilization and reactivation of p53 for targeted cancer therapies.METHODS The cytotoxicity of a potent Cyclophilin A(CypA)inhibitor HL001 was tasted against a panel of cancer cell lines.The genotypes and activation of p53 were compared with the cytotoxicity profile of HL001.Two-dimensional(2D)PAGE analysis was performed to investigate differentially expressed proteins that involves in the anti-proliferation effects of HL001.Pull-down and Co-IP were used to confirmed the new identified PPI between CypA and G3BP1 and orthotopic animal model of lung cancer was used to tested the anti-tumor activity of HL001 in vivo.RESULTS We identify a novel CypA small molecule inhibitor HL001 that induces non-small cell lung cancer(NSCLC)cell cycle arrest and apoptosis via restoring p53 expression.We find that HL001 stabilizes p53 through inhibiting the MDM2-mediated p53 ubiquitination.Further mechanistic studies reveal that the downregulation of G3BP1 and the induction of reactive oxygen species and DNA damage by HL001 contribute to p53 stabilization.Surprisingly,HL001 selectively suppresses tumor growth in p53wildtype NSCLC harboring Arg72 homozygous alleles(p53-72R)through disrupting interaction between MDM2 and p53-72R in a CypA dependent manner.Moreover,combining HL001 with cisplatin synergistically enhance tumor regression in orthotopic NSCLC mouse model.CONCLUSION Pharmacologic inhibition of CypA offers a potential therapeutic strategy via specific activation of p53-72R in NSCLC.展开更多
非小细胞肺癌(non-small cell lung cancer,NSCLC)是全球最常见且致命的恶性肿瘤之一,早期NSCLC以手术切除为主要治疗手段。肿瘤经气腔扩散(spread through air spaces,STAS)是一种肿瘤向肺部气腔扩散的新型传播模式,其与亚肺叶切除术...非小细胞肺癌(non-small cell lung cancer,NSCLC)是全球最常见且致命的恶性肿瘤之一,早期NSCLC以手术切除为主要治疗手段。肿瘤经气腔扩散(spread through air spaces,STAS)是一种肿瘤向肺部气腔扩散的新型传播模式,其与亚肺叶切除术后的发生、复发和远处转移密切相关,使其成为手术方式选择和预后评估中至关重要的考虑因素。STAS阳性患者的术后复发率显著高于STAS阴性患者,其分子机制涉及肿瘤微环境的重塑、特异性基因突变及上皮-间质转化等过程。包括计算机断层扫描(CT)和正电子发射断层扫描/CT在内的影像技术已显示出术前预测STAS的潜力,但其准确性和可操作性仍有待提升。本文综述了STAS的定义、病理特征及相关机制,重点探讨STAS阳性患者的手术方式选择及其在早期NSCLC亚肺叶切除术后癌症复发中的作用,并提出未来研究方向,包括优化STAS的术前诊断方法、探索分子靶向治疗以及建立基于影像学的精准预测模型。展开更多
基金supported by the National Natural Science Foundation of China(81572870)
文摘Objective:To explore the relationship between cancer awareness and the survival of the patients with non-small cell lung carcinoma(NSCLC).Methods:A total of 865 NSCLC patients were screened for the risk factors,including age,gender,address,tumor/lymph nodes/metastasis(TNM)stage,and cancer awareness.Survival of the patients was calculated by Kaplan-Meier method and Cox regression analysis.Results:After an average observation time of 304 d(ranging from 0 to 4718 d),62 of the 394 patients in the cancer awareness group survived,whereas 26 of the 471 patients in the cancer concealment group survived.Cancer-specific and all-cause survival was poorer in the cancer concealment group(P<0.001 for each,log-rank test).Cox multivariate regression analysis showed that cancer concealment displayed significantly lower cancer-specific survival[hazard ratio(HR)=1.534,95%con fi dence interval(CI)1.320 to 1.784,P<0.001]and all-cause survival(HR=1.558,95%CI 1.346 to 1.803,P<0.001).Conclusion:Cancer concealment is associated with a poor survival of NSCLC patients,which may prohibit the patients from obtaining the real“right to survival”.
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.
基金The project supported by FDCT grants from the Science and Technology Development Fund of Macao(021/2013/A1)Scientific Research Project by the Ministry of Science and Technology of Chinathe Macao Science and TechnologyDevelopment Fund(005/2014/AMJ)
文摘OBJECTIVE Lung cancer is the leading cause of cancer death worldwide.Epidermal growth factor receptor(EGFR)mutation(s)is/are common in non-small cell lung cancer(NSCLC)in Asian population,resulting in lung tumor formation.L858 Rsubstitution mutation on exon 21 and in-flame deletion mutation on exon 19 are the two most common forms of EGFR mutation.Molecular targeted therapy using tyrosine kinase inhibitor(TKI)targeting EGFR shows promising initial response,however drug resistance is common.Therefore,it is needed to identify new inhibitors to tackle TKI-resistance.In this study,we aim to investigate the effect of multiple single purified compounds derived from Chinese herbal medicines(CMHs)on a panel of NSCLC cell lines with different EGFR mutational statuses and TKI sensitivity.We also examine the biological functional effect and drug action mechanism of these cell lines after drug treatment.METHODS We have reviewed the literature and selected ten single purified compounds derived from CMHs which exhibited the highest potential of cancer suppression effect in NSCLC.We have recruited three EGFR-dependent NSCLC cell lines for drug screening using cytotoxicity assay.A549 is used as EGFR wild-type control.Two TKI-resistant NSCLC cell lines were used,H1975 harbors double mutation(EGFRL858R+T790M)and H1650 harbors EGFRexon 19 deletion.H2228is a NSCLC cell line which harbours EML4-ALK fusion gene and was used as EGFR-independent cell line control.MTT assay was used to determine the drug efficacy and IC50 value.Then functional assays including cell cycle arrest analysis and apoptosis assay was used to determine the biological effect after drug treatment.RESULTS MTT assay revealed that six out of ten candidate agents showed significant cancer-inhibiting effects in H1650 and H1975cells.Three compounds exhibited IC50 value at micro-molar levels while another three compounds exhibited IC50 at as low as nano-molar levels.One compound exhibited specificity on EGFR-dependent NSCLC cell lines,which showed 10-fold more selective than the EGFR-independent H2228 cells.Cell cycle analysis and immunoblotting assay showed that one effective compound,designated was MUST-1,altered the metabolic pathway of glucose metabolism and lipid metabolism,and induced cell cycle arrest at G1 phase in NSCLC with EGFR mutation.However,the anti-proliferative effects were distinct in NSCLC cell lines with different EGFR mutation patterns.CONCLUSION MUST-1 significantly induced cell cycle arrest in four NSCLC in EGFR mutant cell lines but the inhibiting effect was not significant in EGFR wildtype cell line.Immunobloting assay revealed that overall intracellular lipid content,glycolytic enzymes PKM2 and cell cycle regulatory gene expression were altered after 72 hcompound treatment in the responsive cells.Further investigation is required to elucidate the underlying reason of drug selectivity and the role of glucose and lipid metabolism in regulating drug sensitivity.
基金National Natural Science Foundation of China(81774013,81804221,82074129)and National Science and Technology Major Project of China(2018ZX09721004-006-004)。
文摘OBJECTIVE To investigate the pharmacological effect and mechanism of Sanguisorba officinalis L.(SOL)in non-small cell lung cancer(NSCLC)in vitro and in vivo based on network pharmacology.METHODS Network pharmacology was used to analyze the improving effect of SOL on NSCLC and possible targets.Cell counting kit 8(CCK-8)and 5-ethynyl-2′-deoxyuridine(EdU)staining,Western blotting,flow cytometry of AnnexinⅤ/PI,Hoechst 33342/PI staining detection and immunofluorescence were utilized in vitro.H&E staining,immunohistochemistry staining and Western blotting were performed in vivo.RESULTS Based on network prediction,we analyzed the 208 common targets of SOL and NSCLC.36 core targets in 208 common targets were obtained through cytoscape analysis.And the top 10 core targets included Akt,mTOR,EGFR,etc..KEGG analysis showed that PI3K-Akt signaling pathway was the most likely pathway.Furthermore,the mechanism study found that SOL could activate the PI3K/Akt/mTOR signaling pathway in vitro and in vivo.The anti-proliferative effect of SOL in A549 and H1299 cells was measured and validated by CCK-8 and EdU assay.Immunohistochemical results of Ki67 showed that SOL effectively inhibited tumor growth in vivo.SOL also significantly inhibited the migration and invasion of A549 and H1299 cells.SOL significantly increased the percentage of cells with PI signal in A549 and H1299,and the process of cell death of A549 cells indicated that SOL induced apoptosis.The PARP-1 and caspase-3 in A549 and H1299 were found to be activated in a dose manner.The results in vivo were consistent with those in vitro.CONCLUSION SOL-induced,caspase-3-mediated apoptosis via the induction of the PI3K/Akt/mTOR signaling pathway in NSCLC,which further clarified the mechanism of SOL in the inhibition of NSCLC,and thereby provided a possibility for SOL to serve as a novel therapeutic agent for NSCLC in the future.
基金supported by National Natural Science Foundation of China(31371485,81402482,91313303 and 81573020)CAS Key Laboratory of Receptor Research,the Shanghai Committee of Science and Technology(15431902000)
文摘OBJECTIVE To explored the potential of pharmacological stabilization and reactivation of p53 for targeted cancer therapies.METHODS The cytotoxicity of a potent Cyclophilin A(CypA)inhibitor HL001 was tasted against a panel of cancer cell lines.The genotypes and activation of p53 were compared with the cytotoxicity profile of HL001.Two-dimensional(2D)PAGE analysis was performed to investigate differentially expressed proteins that involves in the anti-proliferation effects of HL001.Pull-down and Co-IP were used to confirmed the new identified PPI between CypA and G3BP1 and orthotopic animal model of lung cancer was used to tested the anti-tumor activity of HL001 in vivo.RESULTS We identify a novel CypA small molecule inhibitor HL001 that induces non-small cell lung cancer(NSCLC)cell cycle arrest and apoptosis via restoring p53 expression.We find that HL001 stabilizes p53 through inhibiting the MDM2-mediated p53 ubiquitination.Further mechanistic studies reveal that the downregulation of G3BP1 and the induction of reactive oxygen species and DNA damage by HL001 contribute to p53 stabilization.Surprisingly,HL001 selectively suppresses tumor growth in p53wildtype NSCLC harboring Arg72 homozygous alleles(p53-72R)through disrupting interaction between MDM2 and p53-72R in a CypA dependent manner.Moreover,combining HL001 with cisplatin synergistically enhance tumor regression in orthotopic NSCLC mouse model.CONCLUSION Pharmacologic inhibition of CypA offers a potential therapeutic strategy via specific activation of p53-72R in NSCLC.
文摘非小细胞肺癌(non-small cell lung cancer,NSCLC)是全球最常见且致命的恶性肿瘤之一,早期NSCLC以手术切除为主要治疗手段。肿瘤经气腔扩散(spread through air spaces,STAS)是一种肿瘤向肺部气腔扩散的新型传播模式,其与亚肺叶切除术后的发生、复发和远处转移密切相关,使其成为手术方式选择和预后评估中至关重要的考虑因素。STAS阳性患者的术后复发率显著高于STAS阴性患者,其分子机制涉及肿瘤微环境的重塑、特异性基因突变及上皮-间质转化等过程。包括计算机断层扫描(CT)和正电子发射断层扫描/CT在内的影像技术已显示出术前预测STAS的潜力,但其准确性和可操作性仍有待提升。本文综述了STAS的定义、病理特征及相关机制,重点探讨STAS阳性患者的手术方式选择及其在早期NSCLC亚肺叶切除术后癌症复发中的作用,并提出未来研究方向,包括优化STAS的术前诊断方法、探索分子靶向治疗以及建立基于影像学的精准预测模型。
