促凋亡基因puma与非小细胞肺癌的相关性研究
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摘要
目的探讨puma基因突变、mRNA及其功能蛋白表达与肺癌的相关性;PUMA与p53的依赖性关系。
方法通过对实验条件的探讨,建立了扩增高GC碱基的PCR方法,使GC含量高达82%的模板成功扩增;建立了实时荧光定量RT-PCR检测puma mRNA的方法。应用所建立的方法和免疫印迹法(Western Blot,WB)检测40例肺癌患者的癌组织、癌旁组织和远癌组织及10例肺良性病变组织中DNA和BH3结构域的突变:puma mRNA及其编码蛋白表达的情况;其次,TUNEL检测其中20例肺癌患者的癌组织、癌旁组织和远癌组织的细胞凋亡状态。
结果
1.通过对实验条件的探讨,确立了puma基因三个外显子的最终扩增条件,分别如下:
(1)外显子2(Exon2)的反应体系为dNTP 200μmol/L,MgSO4 3.0mmol/L,引物500nmol/L,TaqDNA聚合酶0.1U/μL,模板量不低于4 000ng,10×缓冲液5μL,增强液10μL,总体积50μL。反应条件为95℃预变性5min,95℃45Sec、64℃45sec、72℃45sec,循环33次,最后72℃延伸5min。
(2)外显子3(Exon3)的反应体系MgSO4 2.0mmol/L,外显子4(Exon4)的反应体系MgSO4 1.0mmol/L,其余组分和扩增程序与外显子2相同。
2.应用优化后的扩增体系和条件对40例肺癌患者的癌组织,癌旁组织和远癌组织以及10例肺良性病变患者的肺组织的puma基因2、3和4共3个外显子进行了PCR扩增,扩增产物经纯化后测序检测突变及小的删除片段,经Blast 2与GeneBank序列比对,未发现一例外显子突变及小的删除片段,HB3结构域也没有变异。
3.通过对实验条件的探讨,检测puma mRNA实时荧光定量RT-PCR反应体系为cDNA 1.0μl,dNTP 800μmol/L,Taq DNA聚合酶5.0 u,Mg~(2+)1.5mmol/L,1×Enhancersolution 5.0μl,10×buffer 5.0μl,引物各500nmol/L,0.5×SYB Green I 1.0μl,总体积50μl。内参基因GAPDH体系中不加1×Enhancer solution,其余成分同上。扩增条件:94℃预变性5min,94℃变性45see,56℃退火45sec,72℃延伸45sec,共40个循环,终末72℃延伸5 min。
4.应用所建立的实时荧光定量RT-PCR检测puma mRNA表达的相对量,pumamRNA在癌组织、癌旁组、远癌组及肺良性病变对照组中均有表达,癌组织1.17±0.08、癌旁组织1.16±0.07、远癌组织1.18±0.07、肺良性病变对照组织1.17±0.11。经方差分析,puma mRNA的表达在同一病例的癌组织、癌旁组织和远癌组织之间差异无统计学意义(F=0.416,P＞0.05),puma mRNA在肺癌组织和肺良性病变的表达量差别无统计学意义(F=0.264,P＞0.05)。而且鳞癌、腺癌及腺鳞癌的癌组织、癌旁组织和远癌组织中puma mRNA水平差异无统计学意义(F=0.311,P＞0.05;F=0.338,P＞0.05:F=0.329,P＞0.05)。
5.PUMA蛋白检测显示:癌组织、癌旁组织及远癌组织中的表达量分别为1.30(0.98～1.74)、1.11(0.72～1.69)和1.01(0.71～1.49);10份肺良性组织标本的均值为1.17±0.11。经秩和检验分析,在同一病例的癌组织、癌旁组织和远癌组织的PUMA蛋白表达差异无统计学意义(x~2(H)=4.918,P＞0.05);PUMA蛋白在肺癌组织和肺良性病变的表达量差别无统计学意义(x~2(H)=5.092,P＞0.05)。WB结果还显示鳞癌、腺癌及腺鳞癌的癌组织、癌旁组织和远癌组织中PUMA表达水平差异无统计学意义(F=0.101,P＞0.05;F=0.599,P＞0.05;F=1.054,P＞0.05),表明PUMA表达水平与肺癌的病理分型无关。
6.P53蛋白检测显示:癌组织P53蛋白的表达量与癌旁组织、远癌组织和肺良性病变组织之间,蛋白表达差异有统计学意义,癌旁组织和远癌组织之间差异无统计学意义。
7.TUNEL结果显示:癌组织、癌旁组织和远癌组织AI分别是:4.96%±1.63%,1.45%±0.46%和1.26%±0.42%。癌组织的AI高于癌旁组织和远癌组织,AI差异有统计学意义,Z=-0.-3.920,P=0.000;同时癌旁组织与远癌组织之间AI的差异也有统计学意义,Z=-0.-3.469,P=0.001。
8.P53与PUMA关系分析:PUMA蛋白表达与P53蛋白表达之间没有直接关系,其可能的原因是由于肺癌患者P53的突变很高,突变的P53失去来对PUMA的调控作用。
结论
puma在肺癌发生发展过程中基因的结构很稳定,没有发生有意义的突变和BH3结构域的变异;puma mRNA、蛋白表达和细胞凋亡等研究表明,PUMA表达可能不是伴随肺癌发生和发展过程的重要因素;mRNA和蛋白表达水平与肺癌的组织类型无关。P53没有介导PUMA的表达,同时肺癌发生发展的各种因素也没有诱导PUMA的过表达,因此,在肺癌发生发展过程中,细胞凋亡的异常,引起细胞的过度增殖和对放化疗的敏感性减低,导致肺癌一直是全球发病率和死亡率都很高的恶性肿瘤。本研究结果与体外细胞株的研究结果不一致,而与其他肿瘤组织的研究结果一致,说明PUMA在肿瘤的发生发展中,不同的组织有不同的表现,其在肺癌发生发展中的作用还有待进一步的研究,是否能作为肺癌诊断和治疗的新靶点还有待进一步的研究。
Objective to explore the correlation of the mutation and expression of puma gene to tumorigenesis and development of lung cancer and the relation between PUMA and p53.
Methods Explored the condition of PCR and optimized amplification system and reaction conditions of three high GC content Exon of puma,it make the exon 2 that GC content 82%was amplificationed succeefully.Explored the reaction conditions of fluorescence quantitative reverse transcript-polymerase chain reaction,the FQ- RT-PCR assay was established to examine the puma mRNA.The quantitative fluorogenic real-time PCR and Western Blot Analysis were used to detect the mutation of puma,the expression puma mRNA and its protein in all 40 cases of primary lung cancer tissues,para-cancer lung tissues and far-cancer lung tissues,10 control cases of non-cancer lung tissues were also detected. Apoptosis of 20 NSCLC samples were detected by TUNEL technique.
Results
1.By explored the experiment conditions of PCR,the ultimate amplification system and react contions of three exon of puma were eatablished.
(1) The optimized reaction system of exon2 were:dNTP 200μmol/L,enhancer system MgSO4 3.0mmol/L,each primers 500nmol/L,Taq DNA polymerase 0.1U/μl,template more than 4000ng,10×buffer 5μl,ehancer solution 10μl,the total volume was 50μl.The amplificat condition was:After 5 min at 95℃,reactions were cycled through 45s denaturation at 95℃,45s annealing at 64℃,and 45s extension at 72℃for 33 cycles;followed for 5 min.
(2) Exon3 and exon 4 conditions were:MgSO4 2.0mmol/L and 1.0mmol/L,other components were same as exon2.
2.By using the optimized amplification systems and reaction conditions,the exon 2, exon 3 and exon 4 of puma gene in all 40 cases of lung cancer tissues,paracancer lung tissues and farcancer lung tissues,10 cases of lung benign disease were also amplificationed.The amplification productions were depurated and sequenced both mutation and small deleted fragments.The sequencing results compare to GeneBank sequence by Blast 2,non of mutation and small fragments were found,and BH3 domain is normaol.
3.By exploring the conditions of fluorescence quantitative RT-PCR,we showed that the optimized fluorescence quantitative RT-PCR reaction mixtures contained the forward and reverse primers at 500nmol/L each;dNTP 800μmol/L;1.5mmol/L MgCl_2;1×Enhancer solution 5.0μl;10×PCR buffer 5.0μl;0.5×SYB GREEN I 1.0μl;Taq DNA polymerase 0.1U/μl; template 1.0μl;the total reaction volume was 50μl.The RT-PCR was performed under the following conditions:94℃for 5 minutes;followed by 40 cycles of 94℃for 45 seconds,56℃for 45 seconds and 72℃for 45 seconds;and 72℃for 5 minutes.
4.The means of the relative rate of puma mRNA and GAPDH mRNA in lung cancer tissue,paracancer lung tissue,farcancer lung tissue and lung benign disease tissues were 1.17±0.08,1.16±0.07,1.18±0.07 and 1.17±0.11 which detected by fluorescence quantitative RT-PCR.The data were evaluated by analysis of variance.There was no any significant difference among lung cancer tissue,paracancer lung tissue and farcancer lung tissue (F=0.416,P＞0.05).Also,there was no significant difference among squamous cell carcinoma, adenocarcinoma and adenosquamous carcinoma in the group of lung cancer tissue,paracancer lung tissue and farcancer lung tissue(F=0.311,P＞0.05;F=0.338,P＞0.05;F=0.329,P＞0.05).
5.In the 40 cases of primary lung cancer tissues,the medians of PUMA protein in lung cancer tissue,paracancer lung tissue and farcancer lung tissue were 1.30(0.98～1.74), 1.11(0.72～1.69) and 1.01(0.71～1.49) which detected by Western Blot Analysis.The mean of PUMA protein in benigh disease lung tissues was 11.17±0.11 The data were evaluated by analysis of Krusdal-Wallis H test.There were no any significant difference among lung cancer tissue,paracancer lung tissue and farcancer lung tissue(x~2(H) =4.918,P＞0.05).Also,there was no significant difference among squamous cell carcinoma,adenocarcinoma and adenosquamous carcinoma in the group of lung cancer tissue,paracancer lung tissue and farcancer lung tissue(F=0.101,P＞0.05;F=0.599,P＞0.05;F=1.054,P＞0.05).
6.Western Blot Analyzed p53 protein:the p53 protein in cancer tissue is significant difference in paracancer lung tissue,farcancer lung tissue and lung benign disease tissue; there was no any significant difference among paracancer lung tissue and farcancer lung tissue.
7.The results of TUNEL were:The apoptosic index(AI) in lung cancer tissue,paracancer lung tissue and farcancer lung tissue were 4.96%±0.1.63%,1.45%±0.46%and 1.26%±0.42%. AI in lung cancer was significant difference between lung cancer tissue with paracancer lung tissue and farcancer lung tissue(Z=-3.920,P=0.000),AI of paracancer lung tissue was higher than farcancer lung tissue,and has significant difference of them(Z=-3.469,P=0.001).
8 The relationship of PUMA and p53:PUMA protein expression was not correlate with p53 protein expression.The cause of it may be that the p53 has high mutation in lung cancer and the mutated p53 can not imediate expression of PUMA.
Conclusions Mutational analysis revealed that there was no PUMA BH3 domain and exon mutation in the 40 NSCLC,suggesting that PUMA BH3 domain mutation and exon mutation are not a direct target of inactivation in NSCLC development.PUMA expression may be not play a role in NSCLC initiation and progression of cancer.There was not relationship between puma mRNA and protein with different pathological types of NSCLC. PUMA expression was not mediates by p53,and also induced from various apoptotic stimuli. In the course of the occurrence and development of lung cancer,abnormal apoptosis,caused by excessive cell proliferation and to reduce the sensitivity of radiotherapy and chemotherapy, leading to lung cancer has been the global morbidity and mortality are highly malignant.This findings are inconsistent with in vitro cell line findings,but is consistent with other tumor organization findings,explained that PUMA in the tumor occurrence development,different organizations have different performance,and its occurrence in lung cancer development remains to be further research,whether as a lung cancer diagnosis and treatment have yet to be a new target for further study.
方法通过对实验条件的探讨,建立了扩增高GC碱基的PCR方法,使GC含量高达82%的模板成功扩增;建立了实时荧光定量RT-PCR检测puma mRNA的方法。应用所建立的方法和免疫印迹法(Western Blot,WB)检测40例肺癌患者的癌组织、癌旁组织和远癌组织及10例肺良性病变组织中DNA和BH3结构域的突变:puma mRNA及其编码蛋白表达的情况;其次,TUNEL检测其中20例肺癌患者的癌组织、癌旁组织和远癌组织的细胞凋亡状态。
结果
1.通过对实验条件的探讨,确立了puma基因三个外显子的最终扩增条件,分别如下:
(1)外显子2(Exon2)的反应体系为dNTP 200μmol/L,MgSO4 3.0mmol/L,引物500nmol/L,TaqDNA聚合酶0.1U/μL,模板量不低于4 000ng,10×缓冲液5μL,增强液10μL,总体积50μL。反应条件为95℃预变性5min,95℃45Sec、64℃45sec、72℃45sec,循环33次,最后72℃延伸5min。
(2)外显子3(Exon3)的反应体系MgSO4 2.0mmol/L,外显子4(Exon4)的反应体系MgSO4 1.0mmol/L,其余组分和扩增程序与外显子2相同。
2.应用优化后的扩增体系和条件对40例肺癌患者的癌组织,癌旁组织和远癌组织以及10例肺良性病变患者的肺组织的puma基因2、3和4共3个外显子进行了PCR扩增,扩增产物经纯化后测序检测突变及小的删除片段,经Blast 2与GeneBank序列比对,未发现一例外显子突变及小的删除片段,HB3结构域也没有变异。
3.通过对实验条件的探讨,检测puma mRNA实时荧光定量RT-PCR反应体系为cDNA 1.0μl,dNTP 800μmol/L,Taq DNA聚合酶5.0 u,Mg~(2+)1.5mmol/L,1×Enhancersolution 5.0μl,10×buffer 5.0μl,引物各500nmol/L,0.5×SYB Green I 1.0μl,总体积50μl。内参基因GAPDH体系中不加1×Enhancer solution,其余成分同上。扩增条件:94℃预变性5min,94℃变性45see,56℃退火45sec,72℃延伸45sec,共40个循环,终末72℃延伸5 min。
4.应用所建立的实时荧光定量RT-PCR检测puma mRNA表达的相对量,pumamRNA在癌组织、癌旁组、远癌组及肺良性病变对照组中均有表达,癌组织1.17±0.08、癌旁组织1.16±0.07、远癌组织1.18±0.07、肺良性病变对照组织1.17±0.11。经方差分析,puma mRNA的表达在同一病例的癌组织、癌旁组织和远癌组织之间差异无统计学意义(F=0.416,P＞0.05),puma mRNA在肺癌组织和肺良性病变的表达量差别无统计学意义(F=0.264,P＞0.05)。而且鳞癌、腺癌及腺鳞癌的癌组织、癌旁组织和远癌组织中puma mRNA水平差异无统计学意义(F=0.311,P＞0.05;F=0.338,P＞0.05:F=0.329,P＞0.05)。
5.PUMA蛋白检测显示:癌组织、癌旁组织及远癌组织中的表达量分别为1.30(0.98～1.74)、1.11(0.72～1.69)和1.01(0.71～1.49);10份肺良性组织标本的均值为1.17±0.11。经秩和检验分析,在同一病例的癌组织、癌旁组织和远癌组织的PUMA蛋白表达差异无统计学意义(x~2(H)=4.918,P＞0.05);PUMA蛋白在肺癌组织和肺良性病变的表达量差别无统计学意义(x~2(H)=5.092,P＞0.05)。WB结果还显示鳞癌、腺癌及腺鳞癌的癌组织、癌旁组织和远癌组织中PUMA表达水平差异无统计学意义(F=0.101,P＞0.05;F=0.599,P＞0.05;F=1.054,P＞0.05),表明PUMA表达水平与肺癌的病理分型无关。
6.P53蛋白检测显示:癌组织P53蛋白的表达量与癌旁组织、远癌组织和肺良性病变组织之间,蛋白表达差异有统计学意义,癌旁组织和远癌组织之间差异无统计学意义。
7.TUNEL结果显示:癌组织、癌旁组织和远癌组织AI分别是:4.96%±1.63%,1.45%±0.46%和1.26%±0.42%。癌组织的AI高于癌旁组织和远癌组织,AI差异有统计学意义,Z=-0.-3.920,P=0.000;同时癌旁组织与远癌组织之间AI的差异也有统计学意义,Z=-0.-3.469,P=0.001。
8.P53与PUMA关系分析:PUMA蛋白表达与P53蛋白表达之间没有直接关系,其可能的原因是由于肺癌患者P53的突变很高,突变的P53失去来对PUMA的调控作用。
结论
puma在肺癌发生发展过程中基因的结构很稳定,没有发生有意义的突变和BH3结构域的变异;puma mRNA、蛋白表达和细胞凋亡等研究表明,PUMA表达可能不是伴随肺癌发生和发展过程的重要因素;mRNA和蛋白表达水平与肺癌的组织类型无关。P53没有介导PUMA的表达,同时肺癌发生发展的各种因素也没有诱导PUMA的过表达,因此,在肺癌发生发展过程中,细胞凋亡的异常,引起细胞的过度增殖和对放化疗的敏感性减低,导致肺癌一直是全球发病率和死亡率都很高的恶性肿瘤。本研究结果与体外细胞株的研究结果不一致,而与其他肿瘤组织的研究结果一致,说明PUMA在肿瘤的发生发展中,不同的组织有不同的表现,其在肺癌发生发展中的作用还有待进一步的研究,是否能作为肺癌诊断和治疗的新靶点还有待进一步的研究。
Objective to explore the correlation of the mutation and expression of puma gene to tumorigenesis and development of lung cancer and the relation between PUMA and p53.
Methods Explored the condition of PCR and optimized amplification system and reaction conditions of three high GC content Exon of puma,it make the exon 2 that GC content 82%was amplificationed succeefully.Explored the reaction conditions of fluorescence quantitative reverse transcript-polymerase chain reaction,the FQ- RT-PCR assay was established to examine the puma mRNA.The quantitative fluorogenic real-time PCR and Western Blot Analysis were used to detect the mutation of puma,the expression puma mRNA and its protein in all 40 cases of primary lung cancer tissues,para-cancer lung tissues and far-cancer lung tissues,10 control cases of non-cancer lung tissues were also detected. Apoptosis of 20 NSCLC samples were detected by TUNEL technique.
Results
1.By explored the experiment conditions of PCR,the ultimate amplification system and react contions of three exon of puma were eatablished.
(1) The optimized reaction system of exon2 were:dNTP 200μmol/L,enhancer system MgSO4 3.0mmol/L,each primers 500nmol/L,Taq DNA polymerase 0.1U/μl,template more than 4000ng,10×buffer 5μl,ehancer solution 10μl,the total volume was 50μl.The amplificat condition was:After 5 min at 95℃,reactions were cycled through 45s denaturation at 95℃,45s annealing at 64℃,and 45s extension at 72℃for 33 cycles;followed for 5 min.
(2) Exon3 and exon 4 conditions were:MgSO4 2.0mmol/L and 1.0mmol/L,other components were same as exon2.
2.By using the optimized amplification systems and reaction conditions,the exon 2, exon 3 and exon 4 of puma gene in all 40 cases of lung cancer tissues,paracancer lung tissues and farcancer lung tissues,10 cases of lung benign disease were also amplificationed.The amplification productions were depurated and sequenced both mutation and small deleted fragments.The sequencing results compare to GeneBank sequence by Blast 2,non of mutation and small fragments were found,and BH3 domain is normaol.
3.By exploring the conditions of fluorescence quantitative RT-PCR,we showed that the optimized fluorescence quantitative RT-PCR reaction mixtures contained the forward and reverse primers at 500nmol/L each;dNTP 800μmol/L;1.5mmol/L MgCl_2;1×Enhancer solution 5.0μl;10×PCR buffer 5.0μl;0.5×SYB GREEN I 1.0μl;Taq DNA polymerase 0.1U/μl; template 1.0μl;the total reaction volume was 50μl.The RT-PCR was performed under the following conditions:94℃for 5 minutes;followed by 40 cycles of 94℃for 45 seconds,56℃for 45 seconds and 72℃for 45 seconds;and 72℃for 5 minutes.
4.The means of the relative rate of puma mRNA and GAPDH mRNA in lung cancer tissue,paracancer lung tissue,farcancer lung tissue and lung benign disease tissues were 1.17±0.08,1.16±0.07,1.18±0.07 and 1.17±0.11 which detected by fluorescence quantitative RT-PCR.The data were evaluated by analysis of variance.There was no any significant difference among lung cancer tissue,paracancer lung tissue and farcancer lung tissue (F=0.416,P＞0.05).Also,there was no significant difference among squamous cell carcinoma, adenocarcinoma and adenosquamous carcinoma in the group of lung cancer tissue,paracancer lung tissue and farcancer lung tissue(F=0.311,P＞0.05;F=0.338,P＞0.05;F=0.329,P＞0.05).
5.In the 40 cases of primary lung cancer tissues,the medians of PUMA protein in lung cancer tissue,paracancer lung tissue and farcancer lung tissue were 1.30(0.98～1.74), 1.11(0.72～1.69) and 1.01(0.71～1.49) which detected by Western Blot Analysis.The mean of PUMA protein in benigh disease lung tissues was 11.17±0.11 The data were evaluated by analysis of Krusdal-Wallis H test.There were no any significant difference among lung cancer tissue,paracancer lung tissue and farcancer lung tissue(x~2(H) =4.918,P＞0.05).Also,there was no significant difference among squamous cell carcinoma,adenocarcinoma and adenosquamous carcinoma in the group of lung cancer tissue,paracancer lung tissue and farcancer lung tissue(F=0.101,P＞0.05;F=0.599,P＞0.05;F=1.054,P＞0.05).
6.Western Blot Analyzed p53 protein:the p53 protein in cancer tissue is significant difference in paracancer lung tissue,farcancer lung tissue and lung benign disease tissue; there was no any significant difference among paracancer lung tissue and farcancer lung tissue.
7.The results of TUNEL were:The apoptosic index(AI) in lung cancer tissue,paracancer lung tissue and farcancer lung tissue were 4.96%±0.1.63%,1.45%±0.46%and 1.26%±0.42%. AI in lung cancer was significant difference between lung cancer tissue with paracancer lung tissue and farcancer lung tissue(Z=-3.920,P=0.000),AI of paracancer lung tissue was higher than farcancer lung tissue,and has significant difference of them(Z=-3.469,P=0.001).
8 The relationship of PUMA and p53:PUMA protein expression was not correlate with p53 protein expression.The cause of it may be that the p53 has high mutation in lung cancer and the mutated p53 can not imediate expression of PUMA.
Conclusions Mutational analysis revealed that there was no PUMA BH3 domain and exon mutation in the 40 NSCLC,suggesting that PUMA BH3 domain mutation and exon mutation are not a direct target of inactivation in NSCLC development.PUMA expression may be not play a role in NSCLC initiation and progression of cancer.There was not relationship between puma mRNA and protein with different pathological types of NSCLC. PUMA expression was not mediates by p53,and also induced from various apoptotic stimuli. In the course of the occurrence and development of lung cancer,abnormal apoptosis,caused by excessive cell proliferation and to reduce the sensitivity of radiotherapy and chemotherapy, leading to lung cancer has been the global morbidity and mortality are highly malignant.This findings are inconsistent with in vitro cell line findings,but is consistent with other tumor organization findings,explained that PUMA in the tumor occurrence development,different organizations have different performance,and its occurrence in lung cancer development remains to be further research,whether as a lung cancer diagnosis and treatment have yet to be a new target for further study.
引文
[1]JEMALA,SIEGEL R,WARD E,et al.Cancer statistics,2007.CA Cancer J Clin,2007,57(1):43-66
[2]AHMEDIN JEMAL,REBECCA SIEGEL,ELIZABETH WARD,et al.Cancer Statistics,2008.CA Cancer J Clin 2008,58(2):71-96
[3]FERLAY J,BRAYF,PISANI P,et al.GLOBOCAN 2002:Cancer incidence,mortality and prevalence worldwide.CA Cancer J Clin,2005,55(2):74-108
[4]杨玲,李连弟,陈育德,等.中国肺癌死亡趋势分析及发病、死亡的估计与预测[J].中国肺癌杂志,2005,8(4):274-278
[5]YANG L,PARKIN DM,FERLAY J,et al.Estimates of cancer incidence in china for 2000 and projiections for 2005[J].Cancer Epidemiol Biomarkers prev,2005, 14(1):243-250
[6]吕任齐,段体德.云南省九县市肺癌死亡率趋势分析[J].中国肿瘤,2002,11(12):700-701
[7]黄云超,陈小波,张建华,等.1993年至2007年恶性肿瘤住院病人构成分析[J].昆明医学院学报,2008,29(5):89-92
[8]E.STEELS,M.PAESMAN,T.BERGHMANS,et al.Role of p53 as a prognostic factor for survival in lung cancer:a systematic review of the literature with a meta-analysis[J].Eur Respir J,2001,18(4):705-719
[9]JEFFERS JR,PARGANAS E,LEE Y,et al.PUMA is a essential mediator of p53-dependent and independent apoptosis pathways[J].Cancer cell,2003,4(4):321-328
[10]TONG Y,YANG Q,VATER C,et al.The pro-apoptotic protein,Bik,exhibits potent antitumor activity that is dependent on its BH3 domain[J].Mol Cancer Ther,2001,1(2):95-102
[11]ASHKENAZI A,DIXIT VM.Death receptors:signaling and modulation[J].Science,1998,281(5381):1305-1308
[12]GEREN DR,REEDJ C.Mitochondria and apoptosis[J].Science,1998,281(5381):1309-1312
[13]ADAMS JM,CORY S.Life-or-death decisions by the Bcl-2 protein family[J].Ternds Biochem Sci,2001,26(1):61-66
[14]YU J,ZHANG L,HWANG PM,et al.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[15]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].Mol Cell,2001,7(3):683-694
[16]YU J,WANG Z,KINZLER KW,et al.PUMA mediates the apoptotic response to p53in colorectal cancer cells[J].Proc Natl Acad Sci USA,2003,100(4):1931-1936
[17]HAN JW,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[18]CHIPUK JE,BOUCHIER-HAYES L,KUWANA T,et al.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[19]VILLUNGER A,MICHALAK EM,COULTAS L,et al.p53 and drug-induced apoptotic responses mediated by BH3-only proteins puma and noxa[J].Science,2003,302(5647):1036-1038
[1]HOQUE MO,BEGUM S,SOMMER M,et al.PUMA in head and neck cancer[J].Cancer Lett,2003,199(1):75-81
[2]NEWTON C.R,GRAHAM A.PCR[M].1994,Oxford,UK,BIOS Scientific Publishers,Ltd.
[3]POMP D,J.F.MEDRANO.Organic solvents as facilitators of polymerase chain reaction [J].Bio Techniques,1991,10(1):58-59
[1]HOQUE MO,BEGUM S,SOMMER M,et al.PUMA in head and neck cancer[J].Cancer Lett,2003,199(1):75-81
[2]YU J,ZHANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[3]JIAN YU,WEN YUE,BIN WU,et al.PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation[J].Clin Cancer Res,2006,12(9):2928-2936
[4]DING WX,NI HM,CHEN X,et al.A coordinated action of Bax,PUMA,and p53promotes MG132-induced mitochondria activation and apoptosis in colon cancer cells [J].Mol Cancer Ther,2007,6(3):1062-1069
[5]KARST AM,DAI DL,MARTINKA M,et al.PUMA expression is significantly reduced in human cutaneous melanomas[J].Oncogene,2005,24(6):1111-1116
[6]JANSSON A,ARBMAN G,SUN XF,et al.mRNA and protein expression of PUMA in sporadic colorectal cancer[J].Ontology Reports,2004,12(6):1245-1249
[7]江河,江孝清,周绪红,等.喉鳞状细胞癌中PUMA、P53基因的表达及其临床意义[J].武汉大学学报(医学版),2006,27(1):57-59
[8]徐波,王保捷,李爱敏,等.糖皮质激素对急性淋巴细胞白血病Puma表达的影响[J].中国当代儿科杂志,2006,8(2):151-154
[9]郝延璋,叶艳,谢奇朋,等.顺铂诱导凋亡耐受胃腺癌细胞坏死性死亡[J].安徽医科大学学报,2006,41(1):4-6
[10]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].Mol Cell,2001,7(3):683-694
[11]BRAMBILLA E,TRAVIS WD,Colby EV,et al.The new World Health Organization classification of lung tumours[J].Eur Respir J,2001,18(6):1059-1068
[12]YU J,WANG Z,KINZLER KW,et al.PUMA mediates the apoptotie response to p53in colorectal cancer cells[J].Proc Natl Acad Sci USA,2003,100(4):1931-1936
[13]张克君,李德春,朱新国,等.PUMA基因转染对胰腺细胞生长的影响[J].胰腺病学,2006,12(6):355-357
[14]GU J,ZHANG L,SWISHER SG,et al.Induction of p53-regulated genes in lung cancer cells:implications of the mechanism for adenoviral p53-mediated apoptosis[J].Oncogene,2004,23(6):1300-1307
[15]CHANG H.AHN,EUN G.JEONG,SUNG S.KIM,et al.Expressional and mutational analysis of pro-apoptotic Bcl-2 member puma in hepatocellular carcinomas[J].Dig Dis Sci,2007,53(5):1395-1399
[1]JEMAL A,SIEGEL R,WARD E,et al.Cancer statistics,2007.CA Cancer J Clin,2007,57(1):43-66
[2]MARCIN N,JANUSZ A,MADE J,et al.Expression of E-cadherin,β-catenin and Ki-67antigen and their reciprocal relationships in mammary adenocarcinomas in bitches[J].ET Cytobiologica,2007,45(3):233-238
[3]阮永华,金克炜.肺癌浸润前病变及基因改变[J].肿瘤防治杂志,2005,12(15):1189-1193
[4]YU J,ZHANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[5]HAN JW,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[6]HEMANN MT,ZILFOU JT,ZHAO Z,et al.Suppression of tumorigenesis by the p53target PUMA[J].Proc Natl Acad Sci USA,2004,101(25):9333-9338
[7]JANSSON A,ARBMAN G,SUN XF,et al.mRNA and protein expression of PUMA in sporadic eolorectal cancer[J].Ontology Reports,2004,12(6):1245-1249
[8]TRAVIS WD,COLBY TV,CORRIN B,et al.Histological typing of lung and pleural tumours,3rd ed[A].WHO:World Health Organization international histological,Berlin:Springer,1999
[9]FALK OHL,MONIKA JUNG,CHUANLIANG XU,et al.Gene expression studies in prostate cancer tissue:which reference gene should be selected for normalization?[J].MolMed,2005,83(12):1014-1024
[10]SAIKI R,SCHARF S,FALOVNA F,et al.Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia[J].Science,1985,230(4732):1350-1354
[11]BECKER K,PAN D,WHITLEY CB.Real-time quantitative polymerase chain reaction to assess gene transfer.Hum Gene Ther,1999,10(15):2559-2566
[12]WONG ML,MEDRANO JF.Real-time PCR for mRNA quantitation.Biotechniques,2005,39(1):75-85
[13]李光伟,刘和,张峰,等.荧光定量PCR监测五氯酚对好氧颗粒污泥和活性污泥中氨氧化细菌数量的影响[J].微生物学报,2007,47(1):136-140
[14]郝延璋,叶艳,谢奇朋,等.顺铂诱导凋亡耐受胃腺癌细胞坏死性死亡[J].安徽医科大学学报,2006,41(1):4-6
[15]RIZWAN S.AKHTAR,YING GENG,BARBARA J.Klocke,et al.BH3-Only Proapoptotic Bcl-2 Family Members Noxa and Puma Mediate Neural Precursor Cell Death[J].Neurobiology of Disease,2006,26(27):7257-7264
[1]YU J,HANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[2]HAN JW,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[3]CHIPUK JE,BOUCHIER-HAYES L,KUWANA T,ET AL.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[4]JANSSON A,ARBMAN G,SUN XF,et al.mRNA and protein expression of PUMA in sporadic colorectal cancer[J].Ontology Reports,2004,12(6):1245-1249
[5]KARST AM,DAI DL,MARTINKA M,et al.PUMA expression is significantly reduced in human cutaneous melanomas[J].Oncogene,2005,24(6):1111-1116
[6]HEMANN MT,ZILFOU JT,ZHAO Z,et al.Suppression of tumorigenesis by the p53target PUMA[J].Proc Natl Acad Sci USA,2004,101(25):9333-9338
[7]TRAVIS WD,COLBY TV,CORRIN B,et al.Histological typing of lung and pleural tumours,3rd ed[A].WHO:World Health Organization international histological,Berlin:Springer,1999
[8]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].MolCell,2001,7(3):683-694
[9]VILLUNGER A,MICHALAK EM,COULTAS L,et al.p53 and drug-induced apoptotic responses mediated by BH3-only proteins puma and noxa[J].Science,2003,302(5647):1036-1038
[10]JIAN YU,WEN YUE,BIN WU,et al.PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation[J].Clin Cancer Res,2006,12(9):2928-2936
[11]江河,江孝清,周绪红,等.喉鳞状细胞癌中PUMA、P53基因的表达及其临床意义[J].武汉大学学报(医学版),2006,27(1):57-59
[12]徐波,王保捷,李爱敏,等.糖皮质激素对急性淋巴细胞白血病Puma表达的影响[J].中国当代儿科杂志,2006,8(2):151-154
[13]张克君,李德春,朱东明.PUMA蛋白在胰腺癌中的表达及临床意义[J].世界华人消化杂志,2008,16(5):488-492
[14]LA PORTA CA.Drug resistance in melanoma:new perspectives[J].Curr Med Chem,2007,14(4):387-391
[15]ITO H,KANZAWA T,MIYOSHI T,et al.Therapeutic efficacy of PUMA for malignant glioma cells regardless of p53 status[J].Hum Gene Ther,2005,16(6):685-698
[16]GU J,ZHANG L,SWISHER SG,et al.Induction ofp53-regulated genes in lung cancer cells:implications of the mechanism for adenoviral p53-mediated apoptosis[J].Oncogene,2004,23(6):1300-1307
[17]王新颖,何美蓉,李明,等.PUMA在奥沙利铂治疗大肠癌中作用机制的实验研究[J].医学研究生学报,2007,20(4):366-369
[18]吴忧,白澎.p53与人类肺癌关系研究的进展[J].国际呼吸杂志,2006,26(2):99-102
[19]YUAN A,YU CJ,LUH KT,et al.Aberrant p53 expression correlates with expression of vascular endothelial growth factor mRNA and interleuk,in-8 mRNA and neoangiogenesis in non-small-cell lung cancer[J].J Clin Oncol.2002,20(4):900-10
[20]CHOY EY,SIU KL,KOK KH,et al.An Epstein-Barr virus-encoded microRNA targets PUMA to promote host cell survival[J].J Exp Med,2008,205(11):2551-60.
[21]Tsuruya K,Yotsueda H,Ikeda H,et al.Involvement ofp53-transactivated Puma in cisplatin-induced renal tubular cell death.Life Sci,2008,83(15-16):550-556
[22]WIJSMAN JH,JONKE RR,KEIJZER R,et al.A new method to detect apoptosis in paraffin section:in situ end-labeling of fragmented DNA[J].Histochem and Cytochem,1993,41(1):7-12
[23]高金莉,金克伟.Bc1-2,Bax基因与肺癌[J].国外医学生理与病理学临床分册,2001,21(4):256-268
[24]MORI S,ITO G,USAMI N,et al.p53 apoptotic pathway molecules are frequently and simultaneously altered in nonsmall cell lung carcinoma[J].Cancer,2004,100(8):1673-82
[25]MINI(?) V,ANDJELI(?) G,STANI(?) V,et al.p53 and c-myc alterations in patients with non-small cell lung carcinoma in Serbia[J].J BUON,2004,9(1):67-72
[1]FRIDMAN J S,LOWE S W.Control of apoptosis by p53[J].Oncogene,2003,22(56):9030-9040
[2]YU J,ZHANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[3]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].Mol Cell,2001,7(3):683-694
[4]HAN J,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[5]YU J,WANG Z,KINZLER KW,et al.PUMA mediates the apoptotic response to p53 in colorectal cancer cells.Proc Natl Acad Sci USA[J].2003,100(4):1931-1936
[6]CHENG EH,WEI MC,WEILER S,et al.BCL-2,BCL-X(L) sequester BH3 domain-only molecules preventing BAX-and BAK-mediated mitochondrial apoptosis[J].Mol Cell,2001,8(3):705-711
[7]LEWINB.Gene Ⅷ.San Francisco:Benjamin Cummings,2004
[8]CHIPUK JE,BOUCHIER-HAYES L,KUWANA T,et al.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[9]HEMANN MT,ZILFOU JT,ZHAO Z,et al.Suppression of tumorigenesis by the p53 target PUMA[J].Proc Natl Acad Sci U S A,2004,101(25):9333-9338
[10]JEFFERS JR,PARGANAS E,LEE Y,et al.PUMA is an essential mediator of p53-dependent and-independnt apoptotic pathways[J].Cancer Cell,2003,4(4):321-328
[11]CHIPUK J E,BOUCHIER-HAYES L,KUWANA T,et al.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[12]DING WX,NI HM,CHEN X,et al.A coordinated action of Bax,PUMA,and p53 promotes MG132-induced mitochondria activation and apoptosis in colon cancer cells[J].Mol Cancer Ther,2007,6(3):1062-1069
[13]VILLUNGER A,MICHALAK EM,COULTAS L,et al.p53 and drug-induced apoptotic responses mediated by BH3-only proteins puma and noxa[J].Science,2003,302(5647):1036-1038
[14]ERLACHER M,MICHALAK EM,KELLY PN,et al.BH3-only proteins Puma and Bim are rate-limiting for gamma-radiation-and glucocorticoid-induced apoptosis of lymphoid cells in vivo[J].Blood,2005,106(13):4131-4138
[15]SHALTOUKI A,FREER M,MEI Y,et al.Increased expression of the pro-apoptotic Bcl(2) family member PUMA is required for mitochondrial release of cytochrome C and the apoptosis associated with skeletal myoblast differentiation[J].Apoptosis,2007,12(12):2143-2154
[16]HOQUE MO,BEGUM S,SOMMER M,et al.PUMA in head and neck cancer[J].Cancer Lett,2003,199(1):75-81
[17]张克君,李德春,朱东明.PUMA蛋白在胰腺癌中的表达及临床意义[J].世界华人消化杂志,2008,16(5):488-492
[18]CHANG H.AHN,EUN G.JEONG,SUNG S.KIM,et al.Expressional and Mutational Analysis of Pro-apoptotic Bcl-2 Member PUMA in Hepatocellular Carcinomas[J].Dig Dis Sci,2008,53:1395-1399
[19]GU J,ZHANG L,SWISHER SG,et al.Induction of p53-regulated genes in lung cancer cells:implications of the mechanism for adenoviral p53-mediated apoptosis[J].Oncogene,2004,23(6):1300-1307
[20]王新颖,何美蓉,李明,等.PUMA在奥沙利铂治疗大肠癌中作用机制的实验研究[J].医学研究生学报,2007,20(4):366-369
[21]WANG X,LI M,WANG J,et al.The BH3-only protein,PUMA,is involved in oxaliplatin- induced apoptosis in colon cancer cells[J].Biochem Pharmacol,2006,71(11):1540-1545
[22]KARST AM,DAI DL,Martinka M,et al.PUMA expression is significantly reduced in human cutaneous melanomas[J].Oncogene,2005,24(6):1111-1116
[23]ITO H,KANZAWA T,Miyoshi T,et al.Therapeutic efficacy of PUMA for malignant glioma cells regardless of p53 status[J].Hum Gene Ther,2005,16(6):685-698
[24]WANG H,QIAN H,Yu J,et al.Administration of PUMA adenovirus increases the sensitivity of esophageal cancer cells to anticancer drugs[J].Cancer Biol Ther,2006,5(4):380-385
[25]YU J,YUE W,WU B,et al.PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation[J].Clin Cancer Res,2006,12(9):2928-2936
[26]张克君,李德春,朱新国,等.PUMA基因转染对胰腺细胞生长的影响[J].胰腺病学,2006,12(6):355-357
[2]AHMEDIN JEMAL,REBECCA SIEGEL,ELIZABETH WARD,et al.Cancer Statistics,2008.CA Cancer J Clin 2008,58(2):71-96
[3]FERLAY J,BRAYF,PISANI P,et al.GLOBOCAN 2002:Cancer incidence,mortality and prevalence worldwide.CA Cancer J Clin,2005,55(2):74-108
[4]杨玲,李连弟,陈育德,等.中国肺癌死亡趋势分析及发病、死亡的估计与预测[J].中国肺癌杂志,2005,8(4):274-278
[5]YANG L,PARKIN DM,FERLAY J,et al.Estimates of cancer incidence in china for 2000 and projiections for 2005[J].Cancer Epidemiol Biomarkers prev,2005, 14(1):243-250
[6]吕任齐,段体德.云南省九县市肺癌死亡率趋势分析[J].中国肿瘤,2002,11(12):700-701
[7]黄云超,陈小波,张建华,等.1993年至2007年恶性肿瘤住院病人构成分析[J].昆明医学院学报,2008,29(5):89-92
[8]E.STEELS,M.PAESMAN,T.BERGHMANS,et al.Role of p53 as a prognostic factor for survival in lung cancer:a systematic review of the literature with a meta-analysis[J].Eur Respir J,2001,18(4):705-719
[9]JEFFERS JR,PARGANAS E,LEE Y,et al.PUMA is a essential mediator of p53-dependent and independent apoptosis pathways[J].Cancer cell,2003,4(4):321-328
[10]TONG Y,YANG Q,VATER C,et al.The pro-apoptotic protein,Bik,exhibits potent antitumor activity that is dependent on its BH3 domain[J].Mol Cancer Ther,2001,1(2):95-102
[11]ASHKENAZI A,DIXIT VM.Death receptors:signaling and modulation[J].Science,1998,281(5381):1305-1308
[12]GEREN DR,REEDJ C.Mitochondria and apoptosis[J].Science,1998,281(5381):1309-1312
[13]ADAMS JM,CORY S.Life-or-death decisions by the Bcl-2 protein family[J].Ternds Biochem Sci,2001,26(1):61-66
[14]YU J,ZHANG L,HWANG PM,et al.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[15]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].Mol Cell,2001,7(3):683-694
[16]YU J,WANG Z,KINZLER KW,et al.PUMA mediates the apoptotic response to p53in colorectal cancer cells[J].Proc Natl Acad Sci USA,2003,100(4):1931-1936
[17]HAN JW,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[18]CHIPUK JE,BOUCHIER-HAYES L,KUWANA T,et al.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[19]VILLUNGER A,MICHALAK EM,COULTAS L,et al.p53 and drug-induced apoptotic responses mediated by BH3-only proteins puma and noxa[J].Science,2003,302(5647):1036-1038
[1]HOQUE MO,BEGUM S,SOMMER M,et al.PUMA in head and neck cancer[J].Cancer Lett,2003,199(1):75-81
[2]NEWTON C.R,GRAHAM A.PCR[M].1994,Oxford,UK,BIOS Scientific Publishers,Ltd.
[3]POMP D,J.F.MEDRANO.Organic solvents as facilitators of polymerase chain reaction [J].Bio Techniques,1991,10(1):58-59
[1]HOQUE MO,BEGUM S,SOMMER M,et al.PUMA in head and neck cancer[J].Cancer Lett,2003,199(1):75-81
[2]YU J,ZHANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[3]JIAN YU,WEN YUE,BIN WU,et al.PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation[J].Clin Cancer Res,2006,12(9):2928-2936
[4]DING WX,NI HM,CHEN X,et al.A coordinated action of Bax,PUMA,and p53promotes MG132-induced mitochondria activation and apoptosis in colon cancer cells [J].Mol Cancer Ther,2007,6(3):1062-1069
[5]KARST AM,DAI DL,MARTINKA M,et al.PUMA expression is significantly reduced in human cutaneous melanomas[J].Oncogene,2005,24(6):1111-1116
[6]JANSSON A,ARBMAN G,SUN XF,et al.mRNA and protein expression of PUMA in sporadic colorectal cancer[J].Ontology Reports,2004,12(6):1245-1249
[7]江河,江孝清,周绪红,等.喉鳞状细胞癌中PUMA、P53基因的表达及其临床意义[J].武汉大学学报(医学版),2006,27(1):57-59
[8]徐波,王保捷,李爱敏,等.糖皮质激素对急性淋巴细胞白血病Puma表达的影响[J].中国当代儿科杂志,2006,8(2):151-154
[9]郝延璋,叶艳,谢奇朋,等.顺铂诱导凋亡耐受胃腺癌细胞坏死性死亡[J].安徽医科大学学报,2006,41(1):4-6
[10]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].Mol Cell,2001,7(3):683-694
[11]BRAMBILLA E,TRAVIS WD,Colby EV,et al.The new World Health Organization classification of lung tumours[J].Eur Respir J,2001,18(6):1059-1068
[12]YU J,WANG Z,KINZLER KW,et al.PUMA mediates the apoptotie response to p53in colorectal cancer cells[J].Proc Natl Acad Sci USA,2003,100(4):1931-1936
[13]张克君,李德春,朱新国,等.PUMA基因转染对胰腺细胞生长的影响[J].胰腺病学,2006,12(6):355-357
[14]GU J,ZHANG L,SWISHER SG,et al.Induction of p53-regulated genes in lung cancer cells:implications of the mechanism for adenoviral p53-mediated apoptosis[J].Oncogene,2004,23(6):1300-1307
[15]CHANG H.AHN,EUN G.JEONG,SUNG S.KIM,et al.Expressional and mutational analysis of pro-apoptotic Bcl-2 member puma in hepatocellular carcinomas[J].Dig Dis Sci,2007,53(5):1395-1399
[1]JEMAL A,SIEGEL R,WARD E,et al.Cancer statistics,2007.CA Cancer J Clin,2007,57(1):43-66
[2]MARCIN N,JANUSZ A,MADE J,et al.Expression of E-cadherin,β-catenin and Ki-67antigen and their reciprocal relationships in mammary adenocarcinomas in bitches[J].ET Cytobiologica,2007,45(3):233-238
[3]阮永华,金克炜.肺癌浸润前病变及基因改变[J].肿瘤防治杂志,2005,12(15):1189-1193
[4]YU J,ZHANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[5]HAN JW,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[6]HEMANN MT,ZILFOU JT,ZHAO Z,et al.Suppression of tumorigenesis by the p53target PUMA[J].Proc Natl Acad Sci USA,2004,101(25):9333-9338
[7]JANSSON A,ARBMAN G,SUN XF,et al.mRNA and protein expression of PUMA in sporadic eolorectal cancer[J].Ontology Reports,2004,12(6):1245-1249
[8]TRAVIS WD,COLBY TV,CORRIN B,et al.Histological typing of lung and pleural tumours,3rd ed[A].WHO:World Health Organization international histological,Berlin:Springer,1999
[9]FALK OHL,MONIKA JUNG,CHUANLIANG XU,et al.Gene expression studies in prostate cancer tissue:which reference gene should be selected for normalization?[J].MolMed,2005,83(12):1014-1024
[10]SAIKI R,SCHARF S,FALOVNA F,et al.Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia[J].Science,1985,230(4732):1350-1354
[11]BECKER K,PAN D,WHITLEY CB.Real-time quantitative polymerase chain reaction to assess gene transfer.Hum Gene Ther,1999,10(15):2559-2566
[12]WONG ML,MEDRANO JF.Real-time PCR for mRNA quantitation.Biotechniques,2005,39(1):75-85
[13]李光伟,刘和,张峰,等.荧光定量PCR监测五氯酚对好氧颗粒污泥和活性污泥中氨氧化细菌数量的影响[J].微生物学报,2007,47(1):136-140
[14]郝延璋,叶艳,谢奇朋,等.顺铂诱导凋亡耐受胃腺癌细胞坏死性死亡[J].安徽医科大学学报,2006,41(1):4-6
[15]RIZWAN S.AKHTAR,YING GENG,BARBARA J.Klocke,et al.BH3-Only Proapoptotic Bcl-2 Family Members Noxa and Puma Mediate Neural Precursor Cell Death[J].Neurobiology of Disease,2006,26(27):7257-7264
[1]YU J,HANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[2]HAN JW,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[3]CHIPUK JE,BOUCHIER-HAYES L,KUWANA T,ET AL.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[4]JANSSON A,ARBMAN G,SUN XF,et al.mRNA and protein expression of PUMA in sporadic colorectal cancer[J].Ontology Reports,2004,12(6):1245-1249
[5]KARST AM,DAI DL,MARTINKA M,et al.PUMA expression is significantly reduced in human cutaneous melanomas[J].Oncogene,2005,24(6):1111-1116
[6]HEMANN MT,ZILFOU JT,ZHAO Z,et al.Suppression of tumorigenesis by the p53target PUMA[J].Proc Natl Acad Sci USA,2004,101(25):9333-9338
[7]TRAVIS WD,COLBY TV,CORRIN B,et al.Histological typing of lung and pleural tumours,3rd ed[A].WHO:World Health Organization international histological,Berlin:Springer,1999
[8]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].MolCell,2001,7(3):683-694
[9]VILLUNGER A,MICHALAK EM,COULTAS L,et al.p53 and drug-induced apoptotic responses mediated by BH3-only proteins puma and noxa[J].Science,2003,302(5647):1036-1038
[10]JIAN YU,WEN YUE,BIN WU,et al.PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation[J].Clin Cancer Res,2006,12(9):2928-2936
[11]江河,江孝清,周绪红,等.喉鳞状细胞癌中PUMA、P53基因的表达及其临床意义[J].武汉大学学报(医学版),2006,27(1):57-59
[12]徐波,王保捷,李爱敏,等.糖皮质激素对急性淋巴细胞白血病Puma表达的影响[J].中国当代儿科杂志,2006,8(2):151-154
[13]张克君,李德春,朱东明.PUMA蛋白在胰腺癌中的表达及临床意义[J].世界华人消化杂志,2008,16(5):488-492
[14]LA PORTA CA.Drug resistance in melanoma:new perspectives[J].Curr Med Chem,2007,14(4):387-391
[15]ITO H,KANZAWA T,MIYOSHI T,et al.Therapeutic efficacy of PUMA for malignant glioma cells regardless of p53 status[J].Hum Gene Ther,2005,16(6):685-698
[16]GU J,ZHANG L,SWISHER SG,et al.Induction ofp53-regulated genes in lung cancer cells:implications of the mechanism for adenoviral p53-mediated apoptosis[J].Oncogene,2004,23(6):1300-1307
[17]王新颖,何美蓉,李明,等.PUMA在奥沙利铂治疗大肠癌中作用机制的实验研究[J].医学研究生学报,2007,20(4):366-369
[18]吴忧,白澎.p53与人类肺癌关系研究的进展[J].国际呼吸杂志,2006,26(2):99-102
[19]YUAN A,YU CJ,LUH KT,et al.Aberrant p53 expression correlates with expression of vascular endothelial growth factor mRNA and interleuk,in-8 mRNA and neoangiogenesis in non-small-cell lung cancer[J].J Clin Oncol.2002,20(4):900-10
[20]CHOY EY,SIU KL,KOK KH,et al.An Epstein-Barr virus-encoded microRNA targets PUMA to promote host cell survival[J].J Exp Med,2008,205(11):2551-60.
[21]Tsuruya K,Yotsueda H,Ikeda H,et al.Involvement ofp53-transactivated Puma in cisplatin-induced renal tubular cell death.Life Sci,2008,83(15-16):550-556
[22]WIJSMAN JH,JONKE RR,KEIJZER R,et al.A new method to detect apoptosis in paraffin section:in situ end-labeling of fragmented DNA[J].Histochem and Cytochem,1993,41(1):7-12
[23]高金莉,金克伟.Bc1-2,Bax基因与肺癌[J].国外医学生理与病理学临床分册,2001,21(4):256-268
[24]MORI S,ITO G,USAMI N,et al.p53 apoptotic pathway molecules are frequently and simultaneously altered in nonsmall cell lung carcinoma[J].Cancer,2004,100(8):1673-82
[25]MINI(?) V,ANDJELI(?) G,STANI(?) V,et al.p53 and c-myc alterations in patients with non-small cell lung carcinoma in Serbia[J].J BUON,2004,9(1):67-72
[1]FRIDMAN J S,LOWE S W.Control of apoptosis by p53[J].Oncogene,2003,22(56):9030-9040
[2]YU J,ZHANG L,HWANG PM.PUMA induces the rapid apoptosis of colorectal cancer cells[J].Mol Cell,2001,7(3):673-682
[3]NAKANO K,VOUSDEN KH.PUMA,a novel proapoptotic gene,is induced by p53[J].Mol Cell,2001,7(3):683-694
[4]HAN J,FLEMINGTON C,HOUGHTON AB,et al.Expression of bbc3,a pro-apoptotic BH3-only gene,is regulated by diverse cell death and survival signals[J].Proc Natl Acad Sci USA,2001,98(20):11318-11323
[5]YU J,WANG Z,KINZLER KW,et al.PUMA mediates the apoptotic response to p53 in colorectal cancer cells.Proc Natl Acad Sci USA[J].2003,100(4):1931-1936
[6]CHENG EH,WEI MC,WEILER S,et al.BCL-2,BCL-X(L) sequester BH3 domain-only molecules preventing BAX-and BAK-mediated mitochondrial apoptosis[J].Mol Cell,2001,8(3):705-711
[7]LEWINB.Gene Ⅷ.San Francisco:Benjamin Cummings,2004
[8]CHIPUK JE,BOUCHIER-HAYES L,KUWANA T,et al.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[9]HEMANN MT,ZILFOU JT,ZHAO Z,et al.Suppression of tumorigenesis by the p53 target PUMA[J].Proc Natl Acad Sci U S A,2004,101(25):9333-9338
[10]JEFFERS JR,PARGANAS E,LEE Y,et al.PUMA is an essential mediator of p53-dependent and-independnt apoptotic pathways[J].Cancer Cell,2003,4(4):321-328
[11]CHIPUK J E,BOUCHIER-HAYES L,KUWANA T,et al.PUMA couples the nuclear and cytoplasmic proapoptotic function of p53[J].Science,2005,309(5741):1732-1735
[12]DING WX,NI HM,CHEN X,et al.A coordinated action of Bax,PUMA,and p53 promotes MG132-induced mitochondria activation and apoptosis in colon cancer cells[J].Mol Cancer Ther,2007,6(3):1062-1069
[13]VILLUNGER A,MICHALAK EM,COULTAS L,et al.p53 and drug-induced apoptotic responses mediated by BH3-only proteins puma and noxa[J].Science,2003,302(5647):1036-1038
[14]ERLACHER M,MICHALAK EM,KELLY PN,et al.BH3-only proteins Puma and Bim are rate-limiting for gamma-radiation-and glucocorticoid-induced apoptosis of lymphoid cells in vivo[J].Blood,2005,106(13):4131-4138
[15]SHALTOUKI A,FREER M,MEI Y,et al.Increased expression of the pro-apoptotic Bcl(2) family member PUMA is required for mitochondrial release of cytochrome C and the apoptosis associated with skeletal myoblast differentiation[J].Apoptosis,2007,12(12):2143-2154
[16]HOQUE MO,BEGUM S,SOMMER M,et al.PUMA in head and neck cancer[J].Cancer Lett,2003,199(1):75-81
[17]张克君,李德春,朱东明.PUMA蛋白在胰腺癌中的表达及临床意义[J].世界华人消化杂志,2008,16(5):488-492
[18]CHANG H.AHN,EUN G.JEONG,SUNG S.KIM,et al.Expressional and Mutational Analysis of Pro-apoptotic Bcl-2 Member PUMA in Hepatocellular Carcinomas[J].Dig Dis Sci,2008,53:1395-1399
[19]GU J,ZHANG L,SWISHER SG,et al.Induction of p53-regulated genes in lung cancer cells:implications of the mechanism for adenoviral p53-mediated apoptosis[J].Oncogene,2004,23(6):1300-1307
[20]王新颖,何美蓉,李明,等.PUMA在奥沙利铂治疗大肠癌中作用机制的实验研究[J].医学研究生学报,2007,20(4):366-369
[21]WANG X,LI M,WANG J,et al.The BH3-only protein,PUMA,is involved in oxaliplatin- induced apoptosis in colon cancer cells[J].Biochem Pharmacol,2006,71(11):1540-1545
[22]KARST AM,DAI DL,Martinka M,et al.PUMA expression is significantly reduced in human cutaneous melanomas[J].Oncogene,2005,24(6):1111-1116
[23]ITO H,KANZAWA T,Miyoshi T,et al.Therapeutic efficacy of PUMA for malignant glioma cells regardless of p53 status[J].Hum Gene Ther,2005,16(6):685-698
[24]WANG H,QIAN H,Yu J,et al.Administration of PUMA adenovirus increases the sensitivity of esophageal cancer cells to anticancer drugs[J].Cancer Biol Ther,2006,5(4):380-385
[25]YU J,YUE W,WU B,et al.PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation[J].Clin Cancer Res,2006,12(9):2928-2936
[26]张克君,李德春,朱新国,等.PUMA基因转染对胰腺细胞生长的影响[J].胰腺病学,2006,12(6):355-357