Objective Loss-of-function mutation of p53,a tumor suppressor gene,is an important mechanism for the development of human cancers. In this study we tried to transfect p53N15-based fusion peptide into H1299,a lung canc...Objective Loss-of-function mutation of p53,a tumor suppressor gene,is an important mechanism for the development of human cancers. In this study we tried to transfect p53N15-based fusion peptide into H1299,a lung cancer cell line,and evaluate the anti-tumor effects of the fusion peptide. Methods Adeno-associated virus (AAV) vectors were used for transfecting p53N15 fusion peptide into p53-null lung adenocarcinoma H1299 cells.展开更多
Background Patients with single station mediastinal lymph node （N2） non-small call lung ccancer （NSCLC） have a better prognosis than those with multilevel N2.The molecular factors which are involved in disease pro...Background Patients with single station mediastinal lymph node （N2） non-small call lung ccancer （NSCLC） have a better prognosis than those with multilevel N2.The molecular factors which are involved in disease progression remain largely unknown.The purpose of this study was to investigate gene expression differences between single station and multilevel N2 NSCLC and to identify the crucial molecular factors which are associated with progress and prognosis of stage N2 NSCLC.Methods Gene expression analysis was performed using Agilent 4x44K Whole Human Genome Oligo Microarray on 10 freshfrozen lymph node tissue samples from single station N2 and paired multilevel N2 NSCLC patients.Real-time reverse transcription （RT）-PCR was used to validate the differential expression of 14 genes selected by cDNA microarray of which four were confirmed.Immunohistochemical staining for these validated genes was performed on formalin-fixed,paraffinembedded tissue samples from 130 cases of stage N2 NSCLC arranged in a high-density tissue microarray.Results We identified a 14 gene expression signature by comparative analysis of gene expression.Expression of these genes strongly differed between single station and multilevel N2 NSCLC.Four genes （ADAM28,MUC4,CLDN1,and IGF2） correlated with the results of microarray and real-time RT-PCR analysis for the gene-expression data in samples from 56 NSCLC patients.Immunohistochemical staining for these genes in samples from 130 cases of stage N2 NSCLC demonstrated the expression of IGF2 and CLDN1 was negatively correlated with overall survival of stage N2 NSCLC.Conclusions Our results suggest that the expression of CLDN1 and IGF2 indicate a poor prognosis in stage N2 NSCLC.Further,CLDN1 and IGF2 may provide potential targeting opportunities in future therapies.展开更多
Background The critical roles of polyamines in cell growth and differentiation have made polyamJne metabolic pathway a promising target for antitumor therapy. Recent studies have demonstrated in vitro that some antitu...Background The critical roles of polyamines in cell growth and differentiation have made polyamJne metabolic pathway a promising target for antitumor therapy. Recent studies have demonstrated in vitro that some antitumor polyamine analogues could be used as substrates and oxidized by purified recombinant human N^1-acetylpolyamine oxidase （APAO, an enzyme that catabolizes natural poiyamines）, indicating a potential role of APAO in determining the sensitivity of cancer cells to specific antitumor analogues. This study evaluated, in vivo, the effect of APAO on cytotoxicity of antitumor polyamine analogue, N^1-cyclopropylmethyl-N^11-ethylnorspermine （CPENS） and its mechanism when highly expressed in human lung cancer line A549. Methods A clone with high expression of APAO was obtained by transfecting A549 lung cancer ceil line with pcDNA3.1/APAO plasmid and selecting with quantitative realtime PCR and APAO activity assay. Cell proliferation was determined by MTT method and apoptosis related events were evaluated by DNA fragmentation, sub-G1/flow cytometric assay, western blotting （for cytochrome C and Bax） and colorimetric assay （for casapse-3 activity）. Results A clone highly expressing APAO was obtained. High expression of APAO in A549 cells inhibited accumulation of CPENS, decreased their sensitivity to the toxicity of CPENS and prevented CPENS induced apoptosis. Conclusion These results indicate a new drug resisting, mechanism in the tumor cells. High expression of APAO can greatly decrease the sensitivity of tumor cells to the specific polyamine analogues by detoxifying those analogues and prevent analogue induced apoptosis.展开更多
目的 探讨甲状腺激素与N末端脑钠肽前体(NT-proBNP)水平在非小细胞肺癌(NSCLC)患者体内的变化及其临床意义。方法 选取150例经组织病理学确诊的初诊NSCLC患者(观察组)、80例其他癌症初诊患者(其他癌症对照组)和45例体检健康志愿者(健康...目的 探讨甲状腺激素与N末端脑钠肽前体(NT-proBNP)水平在非小细胞肺癌(NSCLC)患者体内的变化及其临床意义。方法 选取150例经组织病理学确诊的初诊NSCLC患者(观察组)、80例其他癌症初诊患者(其他癌症对照组)和45例体检健康志愿者(健康对照组)进行研究。所有受试者空腹采集静脉血,离心后取上清液进行甲状腺激素和NT-proBNP水平的检测,分析各指标在各组的差异。采用Pearson相关性分析进行甲状腺激素与NT-proBNP水平在NSCLC患者中的相关性研究。绘制受试者工作特征曲线(ROC曲线)分析甲状腺激素与NT-proBNP水平对NSCLC患者临床分期和转移情况的诊断价值。结果 观察组患者游离T3(FT3)、游离T4(FT4)、总三碘甲状腺原氨酸(TT3)水平明显低于其他癌症对照组和健康对照组,观察组NT-proBNP水平显著高于其他癌症对照组和健康对照组,其他癌症对照组TT3水平较健康对照组低( P <0.05)。观察组Ⅰ~Ⅱ期和肿瘤无转移患者FT3、FT4、TT3水平明显高于Ⅲ~Ⅳ期和肿瘤有转移者,而NT-proBNP水平呈相反趋势( P <0.05)。Pearson相关分析显示,FT3、FT4、TT3与NT-proBNP水平呈负相关( r =-0.21、-0.163、-0.183,均 P <0.05)。FT3、FT4、TT3、NT-proBNP对NSCLC患者Ⅰ~Ⅱ期和Ⅲ~Ⅳ期临床分期诊断的ROC曲线下面积分别为0.831、0.813、0.713和0.786,灵敏度分别为75.4%、 70.2 %、 73.7 %和63.2%,特异度分别为82.5%、80.7%、63.1%和84.2%。FT3、FT4、TT3、NT-proBNP对NSCLC患者转移情况诊断的ROC曲线下面积分别为0.896、0.714、0.628和0.742,灵敏度分别为80.6%、61.1%、72.2%和63.9%,特异度分别为88.9%、76.4%、79.3%和86.1%。结论 NSCLC患者体内血浆FT3、FT4、TT3水平降低,NT-proBNP水平升高,且与疾病的临床分期和转移情况密切相关。FT3、FT4、TT3水平与NT-proBNP水平间呈负相关,甲状腺激素和NT-proBNP水平对NSCLC患者临床分期和肿瘤转移情况均具有一定的灵敏度和特异度。展开更多
文摘Objective Loss-of-function mutation of p53,a tumor suppressor gene,is an important mechanism for the development of human cancers. In this study we tried to transfect p53N15-based fusion peptide into H1299,a lung cancer cell line,and evaluate the anti-tumor effects of the fusion peptide. Methods Adeno-associated virus (AAV) vectors were used for transfecting p53N15 fusion peptide into p53-null lung adenocarcinoma H1299 cells.
基金Thiswork was supported by grants from National Natural Science Foundation of China （No. 30801377, No. 81000899, No. 81201649） and Tianjin Municipal Science and Technology Commission Key Application Research Project （No. 11JCZDJC 18900）.
文摘Background Patients with single station mediastinal lymph node （N2） non-small call lung ccancer （NSCLC） have a better prognosis than those with multilevel N2.The molecular factors which are involved in disease progression remain largely unknown.The purpose of this study was to investigate gene expression differences between single station and multilevel N2 NSCLC and to identify the crucial molecular factors which are associated with progress and prognosis of stage N2 NSCLC.Methods Gene expression analysis was performed using Agilent 4x44K Whole Human Genome Oligo Microarray on 10 freshfrozen lymph node tissue samples from single station N2 and paired multilevel N2 NSCLC patients.Real-time reverse transcription （RT）-PCR was used to validate the differential expression of 14 genes selected by cDNA microarray of which four were confirmed.Immunohistochemical staining for these validated genes was performed on formalin-fixed,paraffinembedded tissue samples from 130 cases of stage N2 NSCLC arranged in a high-density tissue microarray.Results We identified a 14 gene expression signature by comparative analysis of gene expression.Expression of these genes strongly differed between single station and multilevel N2 NSCLC.Four genes （ADAM28,MUC4,CLDN1,and IGF2） correlated with the results of microarray and real-time RT-PCR analysis for the gene-expression data in samples from 56 NSCLC patients.Immunohistochemical staining for these genes in samples from 130 cases of stage N2 NSCLC demonstrated the expression of IGF2 and CLDN1 was negatively correlated with overall survival of stage N2 NSCLC.Conclusions Our results suggest that the expression of CLDN1 and IGF2 indicate a poor prognosis in stage N2 NSCLC.Further,CLDN1 and IGF2 may provide potential targeting opportunities in future therapies.
基金This work was supported by grants from the National Natural Science Foundation of China （No. 30772590） and Natural Science Foundation of Hubei Department of Education （No. D200713003）.
文摘Background The critical roles of polyamines in cell growth and differentiation have made polyamJne metabolic pathway a promising target for antitumor therapy. Recent studies have demonstrated in vitro that some antitumor polyamine analogues could be used as substrates and oxidized by purified recombinant human N^1-acetylpolyamine oxidase （APAO, an enzyme that catabolizes natural poiyamines）, indicating a potential role of APAO in determining the sensitivity of cancer cells to specific antitumor analogues. This study evaluated, in vivo, the effect of APAO on cytotoxicity of antitumor polyamine analogue, N^1-cyclopropylmethyl-N^11-ethylnorspermine （CPENS） and its mechanism when highly expressed in human lung cancer line A549. Methods A clone with high expression of APAO was obtained by transfecting A549 lung cancer ceil line with pcDNA3.1/APAO plasmid and selecting with quantitative realtime PCR and APAO activity assay. Cell proliferation was determined by MTT method and apoptosis related events were evaluated by DNA fragmentation, sub-G1/flow cytometric assay, western blotting （for cytochrome C and Bax） and colorimetric assay （for casapse-3 activity）. Results A clone highly expressing APAO was obtained. High expression of APAO in A549 cells inhibited accumulation of CPENS, decreased their sensitivity to the toxicity of CPENS and prevented CPENS induced apoptosis. Conclusion These results indicate a new drug resisting, mechanism in the tumor cells. High expression of APAO can greatly decrease the sensitivity of tumor cells to the specific polyamine analogues by detoxifying those analogues and prevent analogue induced apoptosis.
文摘目的 探讨甲状腺激素与N末端脑钠肽前体(NT-proBNP)水平在非小细胞肺癌(NSCLC)患者体内的变化及其临床意义。方法 选取150例经组织病理学确诊的初诊NSCLC患者(观察组)、80例其他癌症初诊患者(其他癌症对照组)和45例体检健康志愿者(健康对照组)进行研究。所有受试者空腹采集静脉血,离心后取上清液进行甲状腺激素和NT-proBNP水平的检测,分析各指标在各组的差异。采用Pearson相关性分析进行甲状腺激素与NT-proBNP水平在NSCLC患者中的相关性研究。绘制受试者工作特征曲线(ROC曲线)分析甲状腺激素与NT-proBNP水平对NSCLC患者临床分期和转移情况的诊断价值。结果 观察组患者游离T3(FT3)、游离T4(FT4)、总三碘甲状腺原氨酸(TT3)水平明显低于其他癌症对照组和健康对照组,观察组NT-proBNP水平显著高于其他癌症对照组和健康对照组,其他癌症对照组TT3水平较健康对照组低( P <0.05)。观察组Ⅰ~Ⅱ期和肿瘤无转移患者FT3、FT4、TT3水平明显高于Ⅲ~Ⅳ期和肿瘤有转移者,而NT-proBNP水平呈相反趋势( P <0.05)。Pearson相关分析显示,FT3、FT4、TT3与NT-proBNP水平呈负相关( r =-0.21、-0.163、-0.183,均 P <0.05)。FT3、FT4、TT3、NT-proBNP对NSCLC患者Ⅰ~Ⅱ期和Ⅲ~Ⅳ期临床分期诊断的ROC曲线下面积分别为0.831、0.813、0.713和0.786,灵敏度分别为75.4%、 70.2 %、 73.7 %和63.2%,特异度分别为82.5%、80.7%、63.1%和84.2%。FT3、FT4、TT3、NT-proBNP对NSCLC患者转移情况诊断的ROC曲线下面积分别为0.896、0.714、0.628和0.742,灵敏度分别为80.6%、61.1%、72.2%和63.9%,特异度分别为88.9%、76.4%、79.3%和86.1%。结论 NSCLC患者体内血浆FT3、FT4、TT3水平降低,NT-proBNP水平升高,且与疾病的临床分期和转移情况密切相关。FT3、FT4、TT3水平与NT-proBNP水平间呈负相关,甲状腺激素和NT-proBNP水平对NSCLC患者临床分期和肿瘤转移情况均具有一定的灵敏度和特异度。
