The genomic fusions of the anaplastic lymphoma kinase(ALK)gene have been widely recognized as effective therapeutic targets for non-small cell lung carcinoma(NSCLC).The Second Xiangya Hospital of Central South Univers...The genomic fusions of the anaplastic lymphoma kinase(ALK)gene have been widely recognized as effective therapeutic targets for non-small cell lung carcinoma(NSCLC).The Second Xiangya Hospital of Central South University has treated 2 NSCLC patients with 2 distinct novel ALK gene fusions.Case 1 was a 55-year-old male with a solid nodule located in the right hilar lobe on enhanced CT scan.Case 2 was a 47-year-old female with enhanced CT showing involvement of the left upper lobe of lung.Histopathological examination of tumor tissues confirmed lung adenocarcinoma in both cases.Immunohistochemical(IHC)staining demonstrated positivity for thyroid transcription factor 1(TTF-1)and ALK-D5F3 in tumor cells,while negativity for P40.The next-generation sequencing(NGS)tests identified a PNPT1-ALK(Exon22:Exon20)fusion variant in case 1 and a TCEAL2-ALK(Exon3:Exon19)fusion variant in case 2.The TCEAL2-ALK fusion was further confirmed by amplification refractory mutation system(ARMS)-PCR at the mRNA level.Both patients were treated with oral alectinib at a dosage of 600 mg twice daily.The tumors in both patients were significantly decreased after alectinib treatment,achieving partial response.At the time of submission,there was an absence of disease progression and the progression-free survival(PFS)had surpassed 1 year.It offered compelling evidences that the individuals with NSCLC and harboring either a PNPT1-ALK(Exon22:Exon20)fusion or a TCEAL2-ALK(Exon3:Exon19)fusion,experience favorable therapeutic outcomes through the administration of alectinib.This study expands the known ALK fusion variants database and supports the precision treatment of NSCLC using ALK tyrosine kinase inhibitors(TKIs).展开更多
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
基金supported by the National Natural Science Foundation(81900070)the Natural Science Foundation of Hunan Province(2020JJ5813)China。
文摘The genomic fusions of the anaplastic lymphoma kinase(ALK)gene have been widely recognized as effective therapeutic targets for non-small cell lung carcinoma(NSCLC).The Second Xiangya Hospital of Central South University has treated 2 NSCLC patients with 2 distinct novel ALK gene fusions.Case 1 was a 55-year-old male with a solid nodule located in the right hilar lobe on enhanced CT scan.Case 2 was a 47-year-old female with enhanced CT showing involvement of the left upper lobe of lung.Histopathological examination of tumor tissues confirmed lung adenocarcinoma in both cases.Immunohistochemical(IHC)staining demonstrated positivity for thyroid transcription factor 1(TTF-1)and ALK-D5F3 in tumor cells,while negativity for P40.The next-generation sequencing(NGS)tests identified a PNPT1-ALK(Exon22:Exon20)fusion variant in case 1 and a TCEAL2-ALK(Exon3:Exon19)fusion variant in case 2.The TCEAL2-ALK fusion was further confirmed by amplification refractory mutation system(ARMS)-PCR at the mRNA level.Both patients were treated with oral alectinib at a dosage of 600 mg twice daily.The tumors in both patients were significantly decreased after alectinib treatment,achieving partial response.At the time of submission,there was an absence of disease progression and the progression-free survival(PFS)had surpassed 1 year.It offered compelling evidences that the individuals with NSCLC and harboring either a PNPT1-ALK(Exon22:Exon20)fusion or a TCEAL2-ALK(Exon3:Exon19)fusion,experience favorable therapeutic outcomes through the administration of alectinib.This study expands the known ALK fusion variants database and supports the precision treatment of NSCLC using ALK tyrosine kinase inhibitors(TKIs).
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.
文摘背景与目的肺癌是全球癌症死亡的主要原因之一,约80%的肺癌属于非小细胞肺癌(non-small cell lung cancer,NSCLC),其中肺鳞癌(lung squamous cell carcinoma,LUSC)在NSCLC中占据重要比例。尽管肿瘤的综合治疗极大提升了患者的总生存期,但晚期LUSC患者的预后较差。急需一种生物标志物来预测晚期LUSC患者的预后,以便通过早期诊断,改善预后。研究发现miRNAs在肺癌组织中差异表达,并作为潜在的致癌或抑癌基因发挥作用,本研究旨在筛选出早期和晚期LUSC差异表达的miRNAs,构建用于预测晚期LUSC患者预后的一组miRNAs标志物。方法从癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中下载LUSC患者临床信息及miRNAs的相关数据。应用生物信息学方法分析数据,绘制受试者工作特征(receiver operating characteristic,ROC)曲线,利用多种在线分析工具预测靶基因,分析靶基因的潜在生物学机制。结果两组间共鉴定出58个差异表达的miRNAs。根据LASSO回归筛选出7个miRNAs拟构建miRNAs标志物,又根据每个miRNAs的ROC曲线下面积(area under the curve,AUC)值,最终选取其中的4个mRNAs(miR-377-3p、miR-4779、miR-6803-5p、miR-3960)作为预测晚期LUSC患者的生物标志物。4个miRNAs联合的AUC值为0.865。富集分析显示这些靶基因富集在癌症通路、促分裂素原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路、丝氨酸/苏氨酸激酶(serine/threonine kinase,STK)及酪氨酸激酶信号通路等多种通路。结论miR-377-3p、miR-4779、miR-6803-5p、miR-3960联合预测晚期LUSC患者预后能力良好,AUC可达0.865。
