Objective:To investigate the potential of HS-10182,a second-generation epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI),as a radiosensitizer in non-small cell lung cancer(NSCLC).Methods:Two cell li...Objective:To investigate the potential of HS-10182,a second-generation epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI),as a radiosensitizer in non-small cell lung cancer(NSCLC).Methods:Two cell lines of NSCLCs,A549 that possesses wild-type(WT)EGFRs and H1975 that possesses EGFR L858R/T790M double mutations,were treated with HS-10182 at various concentrations,and cell viabilities were determined using the MTS assay.The cells were tested by clonogenic survival assays to identify the radiosensitivity of both groups.Western blot was performed to analyze the expression of phosphorylated EGFR,AKT,DNA-dependent protein kinase,and catalytic subunit(DNA-PKcs)proteins.Immunofluorescence analyses were performed to examine the formation and changes in nuclearγ-H2AX foci.Cell apoptosis was examined by flow cytometry and Western blots for cleaved caspase-3,-8,-9,and cleaved poly ADP-ribose polymerase(PARP).Furthermore,we established xenograft models in mice and the effects of different treatments on tumor growth were then assessed.Results:Clonogenic survival assays revealed that HS-10182 significantly enhanced the radiosensitivity of H1975 cells but not A549cells[dose enhancement ratios(DERs)=2.36(P<0.05)vs.1.43(P>0.05)].Western blot results showed that HS-10182 increased the levels of cleaved caspase-3,-8,-9,and cleaved PARP in H1975 cells but not in A549 cells.In addition,flow cytometry analysis showed that HS-10182 enhanced irradiation-induced apoptosis in H1975.Immunofluorescence results found that HS-10182increased the average number ofγ-H2AX foci after irradiation in H1975 cells,but not in A549 cells.Combined radiation and HS-10182 treatment increased the expression of DNA-PKcs but this increase was more significant in H1975 cells than in A549 cells.Moreover,HS-10182 suppressed the increased expression of Rad50 in H1975 cells in response to irradiation.In vivo experiments found that the combined therapy significantly inhibited tumor growth.Conclusions:HS-10182 enhances the radiosensitivity of H1975 cells which is possibly because that HS-10182 could enhance irradiation-induced apoptosis,increase irradiation-induced DNA damage,and cause a delay in DNA damage repair.Our findings suggest that radiotherapy combined HS-10182 is a novel treatment for lung cancer cells which have acquired the T790M mutation.HS-10182 could be brought to the clinic as a radiosensitizer in NSCLCs with the EGFR T790M mutation.展开更多
BACKGROUND The emergence of secondary drug resistance when treating epidermal growth factor receptor(EGFR)mutated non-small cell lung cancer(NSCLC)using EGFRtyrosine kinase inhibitors(EGFR-TKIs),seriously affects the ...BACKGROUND The emergence of secondary drug resistance when treating epidermal growth factor receptor(EGFR)mutated non-small cell lung cancer(NSCLC)using EGFRtyrosine kinase inhibitors(EGFR-TKIs),seriously affects the therapeutic efficacy and survival of patients.Here,we report a case of advanced NSCLC focusing on the application of multiple biopsy modalities to reveal the development of multiple resistance mechanisms during targeted therapies.CASE SUMMARY A 54-year-old male patient presented with EGFR 19Del-mutated advanced lung adenocarcinoma,and exhibited the development of a T790M mutation during initial TKI treatment.Following 3 mo of Osimertinib treatment,a mixed response was observed.Tissue biopsy of the progressive lesion showed transformation to small cell lung cancer(SCLC)harboring RB1 and TP53 mutations,with loss of the original T790M mutation.A standard chemotherapy regimen with Anlotinib for SCLC was administered.Repeat biopsy revealed adenocarcinoma combined with SCLC after tumor progression.The patient’s overall survival was 24 mo.CONCLUSION Multiple biopsy modalities can reveal the development of multiple resistance mechanisms which help with treatment decision-making.Comprehensive treatment regimens according to the drug resistance mechanism significantly improved the prognosis of such patients.展开更多
背景与目的奥希替尼(Osimertinib)是美国食品和药物管理局(Food and Drug Administration,FDA)批准用于携带表皮生长因子受体(epidermal growth factor receptor,EGFR)-T790M突变的晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)...背景与目的奥希替尼(Osimertinib)是美国食品和药物管理局(Food and Drug Administration,FDA)批准用于携带表皮生长因子受体(epidermal growth factor receptor,EGFR)-T790M突变的晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的药物,用药前需行EGFR-T790M检测。不少患者因进展病灶隐匿或体弱无法进行组织活检错过Osimertinib治疗,本研究希望能从血清中筛查出预测EGFR-T790M耐药突变相关蛋白,为临床用药提供帮助。本研究旨在探索EGFR-T790M耐药基因相关蛋白,为临床用药提供帮助。方法本研究纳入口服易瑞沙晚期肺腺癌患者36例,在疾病进展后行组织活检,使用ARMS方法检测出EGFR-T790M突变组患者18例,非EGFR-T790M突变组18例。收集耐药患者血清,采用同位素标记相对和绝对定量标记结合二维液相色谱串联质谱蛋白组学技术筛选与EGFR-T790M耐药相关蛋白。结果筛出17种差异性蛋白,与EGFR-T790M基因突变相关上调蛋白6种,下调蛋白11种,主要参与31种生物过程,7种细胞组分,26种分子功能;反应途径中共鉴定出12种富集途径,其中富集指数最高的是凝血级联途径。结论发现与EGFR-T790M耐药相关蛋白共17种,参与凝血级联途径蛋白有望成为预测EGFRT790M耐药突变相关的生物标志物。展开更多
BACKGROUND Lung squamous cell cancer(LSCC)rarely harbors epidermal growth factor receptor(EGFR)mutations,even much rarer for acquired T790M mutation.Although clinical trials of AURA series illustrated that non-small c...BACKGROUND Lung squamous cell cancer(LSCC)rarely harbors epidermal growth factor receptor(EGFR)mutations,even much rarer for acquired T790M mutation.Although clinical trials of AURA series illustrated that non-small cell lung cancer(NSCLC)with EGFR T790M mutation can benefit from osimertinib,only five LSCC patients were enrolled in total;moreover,the efficacy for LSCC was not shown in the results.Therefore,the response of LSCC to osimertinib is still unclear to date.CASE SUMMARY We report an LSCC case with T790M-related acquired resistance after treatments with first-generation EGFR-tyrosine kinase inhibitors(EGFR-TKIs)and benefited from osimertinib significantly.A 63-year-old Chinese man was diagnosed with stage IV(cT2N2M1b)LSCC harboring an EGFR exon 19-deletion mutation.Following disease progression after gefitinib and multi-line chemotherapy,rebiopsy was conducted.Molecular testing of EGFR by amplification refractory mutation system-polymerase chain reaction detected the exon 19-deletion without T790M mutation.Therefore,the patient was given erlotinib,but progression developed only 3 mo later.Then the frozen re-biopsy tissue was tested by next-generation sequencing(NGS),which detected an EGFR T790M mutation.However,he was very weak with symptoms of dysphagia and cachexia.Fortunately,osimertinib was started,leading to alleviation from the symptoms.Four months later,normal deglutition was restored and partial response was achieved.Finally,the patient achieved an overall survival time period of 29 mo.CONCLUSION Our findings highlight that EGFR T790M mutation may also be an important acquired drug resistance mechanism for LSCC and offer direct evidence of the efficacy of osimertinib in LSCC with T790M mutation.NGS and better preservation conditions may contribute to higher sensitivity of EGFR T790M detection.展开更多
基金supported by the National Natural Science Foundation of China (Grant No. 81372518)the Tianjin Key Problem Tackling Project for Cancer Therapy (Grant No. 12ZCDZSY15900)
文摘Objective:To investigate the potential of HS-10182,a second-generation epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI),as a radiosensitizer in non-small cell lung cancer(NSCLC).Methods:Two cell lines of NSCLCs,A549 that possesses wild-type(WT)EGFRs and H1975 that possesses EGFR L858R/T790M double mutations,were treated with HS-10182 at various concentrations,and cell viabilities were determined using the MTS assay.The cells were tested by clonogenic survival assays to identify the radiosensitivity of both groups.Western blot was performed to analyze the expression of phosphorylated EGFR,AKT,DNA-dependent protein kinase,and catalytic subunit(DNA-PKcs)proteins.Immunofluorescence analyses were performed to examine the formation and changes in nuclearγ-H2AX foci.Cell apoptosis was examined by flow cytometry and Western blots for cleaved caspase-3,-8,-9,and cleaved poly ADP-ribose polymerase(PARP).Furthermore,we established xenograft models in mice and the effects of different treatments on tumor growth were then assessed.Results:Clonogenic survival assays revealed that HS-10182 significantly enhanced the radiosensitivity of H1975 cells but not A549cells[dose enhancement ratios(DERs)=2.36(P<0.05)vs.1.43(P>0.05)].Western blot results showed that HS-10182 increased the levels of cleaved caspase-3,-8,-9,and cleaved PARP in H1975 cells but not in A549 cells.In addition,flow cytometry analysis showed that HS-10182 enhanced irradiation-induced apoptosis in H1975.Immunofluorescence results found that HS-10182increased the average number ofγ-H2AX foci after irradiation in H1975 cells,but not in A549 cells.Combined radiation and HS-10182 treatment increased the expression of DNA-PKcs but this increase was more significant in H1975 cells than in A549 cells.Moreover,HS-10182 suppressed the increased expression of Rad50 in H1975 cells in response to irradiation.In vivo experiments found that the combined therapy significantly inhibited tumor growth.Conclusions:HS-10182 enhances the radiosensitivity of H1975 cells which is possibly because that HS-10182 could enhance irradiation-induced apoptosis,increase irradiation-induced DNA damage,and cause a delay in DNA damage repair.Our findings suggest that radiotherapy combined HS-10182 is a novel treatment for lung cancer cells which have acquired the T790M mutation.HS-10182 could be brought to the clinic as a radiosensitizer in NSCLCs with the EGFR T790M mutation.
文摘BACKGROUND The emergence of secondary drug resistance when treating epidermal growth factor receptor(EGFR)mutated non-small cell lung cancer(NSCLC)using EGFRtyrosine kinase inhibitors(EGFR-TKIs),seriously affects the therapeutic efficacy and survival of patients.Here,we report a case of advanced NSCLC focusing on the application of multiple biopsy modalities to reveal the development of multiple resistance mechanisms during targeted therapies.CASE SUMMARY A 54-year-old male patient presented with EGFR 19Del-mutated advanced lung adenocarcinoma,and exhibited the development of a T790M mutation during initial TKI treatment.Following 3 mo of Osimertinib treatment,a mixed response was observed.Tissue biopsy of the progressive lesion showed transformation to small cell lung cancer(SCLC)harboring RB1 and TP53 mutations,with loss of the original T790M mutation.A standard chemotherapy regimen with Anlotinib for SCLC was administered.Repeat biopsy revealed adenocarcinoma combined with SCLC after tumor progression.The patient’s overall survival was 24 mo.CONCLUSION Multiple biopsy modalities can reveal the development of multiple resistance mechanisms which help with treatment decision-making.Comprehensive treatment regimens according to the drug resistance mechanism significantly improved the prognosis of such patients.
文摘背景与目的奥希替尼(Osimertinib)是美国食品和药物管理局(Food and Drug Administration,FDA)批准用于携带表皮生长因子受体(epidermal growth factor receptor,EGFR)-T790M突变的晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的药物,用药前需行EGFR-T790M检测。不少患者因进展病灶隐匿或体弱无法进行组织活检错过Osimertinib治疗,本研究希望能从血清中筛查出预测EGFR-T790M耐药突变相关蛋白,为临床用药提供帮助。本研究旨在探索EGFR-T790M耐药基因相关蛋白,为临床用药提供帮助。方法本研究纳入口服易瑞沙晚期肺腺癌患者36例,在疾病进展后行组织活检,使用ARMS方法检测出EGFR-T790M突变组患者18例,非EGFR-T790M突变组18例。收集耐药患者血清,采用同位素标记相对和绝对定量标记结合二维液相色谱串联质谱蛋白组学技术筛选与EGFR-T790M耐药相关蛋白。结果筛出17种差异性蛋白,与EGFR-T790M基因突变相关上调蛋白6种,下调蛋白11种,主要参与31种生物过程,7种细胞组分,26种分子功能;反应途径中共鉴定出12种富集途径,其中富集指数最高的是凝血级联途径。结论发现与EGFR-T790M耐药相关蛋白共17种,参与凝血级联途径蛋白有望成为预测EGFRT790M耐药突变相关的生物标志物。
基金Supported by the National Natural Science Foundation of China,No.81402561
文摘BACKGROUND Lung squamous cell cancer(LSCC)rarely harbors epidermal growth factor receptor(EGFR)mutations,even much rarer for acquired T790M mutation.Although clinical trials of AURA series illustrated that non-small cell lung cancer(NSCLC)with EGFR T790M mutation can benefit from osimertinib,only five LSCC patients were enrolled in total;moreover,the efficacy for LSCC was not shown in the results.Therefore,the response of LSCC to osimertinib is still unclear to date.CASE SUMMARY We report an LSCC case with T790M-related acquired resistance after treatments with first-generation EGFR-tyrosine kinase inhibitors(EGFR-TKIs)and benefited from osimertinib significantly.A 63-year-old Chinese man was diagnosed with stage IV(cT2N2M1b)LSCC harboring an EGFR exon 19-deletion mutation.Following disease progression after gefitinib and multi-line chemotherapy,rebiopsy was conducted.Molecular testing of EGFR by amplification refractory mutation system-polymerase chain reaction detected the exon 19-deletion without T790M mutation.Therefore,the patient was given erlotinib,but progression developed only 3 mo later.Then the frozen re-biopsy tissue was tested by next-generation sequencing(NGS),which detected an EGFR T790M mutation.However,he was very weak with symptoms of dysphagia and cachexia.Fortunately,osimertinib was started,leading to alleviation from the symptoms.Four months later,normal deglutition was restored and partial response was achieved.Finally,the patient achieved an overall survival time period of 29 mo.CONCLUSION Our findings highlight that EGFR T790M mutation may also be an important acquired drug resistance mechanism for LSCC and offer direct evidence of the efficacy of osimertinib in LSCC with T790M mutation.NGS and better preservation conditions may contribute to higher sensitivity of EGFR T790M detection.
