APE1在非小细胞肺癌的表达及其与铂类化疗敏感性的关系
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摘要
非小细胞肺癌(Non-small-cell lung cancer,NSCLC)约占肺癌的80%,已经成为全球第一癌症杀手。晚期NSCLC的化疗主要是以铂类为主,多采用铂类加泰素(paclitaxel),泰素帝(docetaxel),健择(gemcitabine)或去甲长春花碱(NVB)为主的联合化疗方案。但是这几种联合化疗方案临床疗效并没有显著差异,表明以铂类为主的化疗目前已经达到瓶颈。为了突破NSCLC的化疗瓶颈,进一步探讨铂类抗癌作用机制及其耐药机制具有非常重要的临床意义。研究表明,铂类抗癌作用的主要靶点为DNA,阻止其复制及转录,造成细胞死亡。铂类耐药的机制仍未完全阐明,可能是由多种因素引起,包括药物积聚减少;药物解毒增加(如谷胱甘肽和金属硫蛋白);DNA修复能力增强及顺铂-DNA加合物增加等。
DNA损伤修复系统作为机体抵抗各种损伤的分子基础,对于维护基因组的稳定与完整起着至关重要的作用,然而对于通过损伤DNA杀死癌细胞的铂类化疗,这一过程无疑削弱了治疗效果。因此,DNA修复能力增强可能是铂类耐药的主要机制之一。DNA损伤修复基因脱嘌呤/脱嘧啶核酸内切酶(apurinic/apyrimidinic endonuclease,AP,1具有核酸内切酶及氧化还原双功能,是细胞放射性损伤和基因毒性药物烷化剂致伤的重要修复因子,并通过氧化还原机制调节多种转录因子及下游靶基因表达,而这些转录因子与肿瘤放化疗抵抗密切相关。因此本研究检测NSCLC患者APE1蛋白的表达特点,分析APEl与NSCLC术后复发和铂类化疗敏感性的关系,并用Ad5/F35-APE1 siRNA重组腺病毒载体特异性“敲除”APE1基因表达,研究其体外对NSCLC细胞顺铂化疗敏感性的影响。
研究目的:
1、研究APE1蛋白在非小细胞肺癌组织细胞中表达情况,分析APE1表达特点与NSCLC术后复发和铂类抵抗的关系,以确证APE1为NSCLC治疗有效的靶基因。
2、探讨诱导APE1基因沉默对NSCLC含铂类化疗的增敏作用。
研究内容和方法:
1、APE1蛋白在NSCLC细胞中的表达特点及NSCLC术后复发和含铂类药物抵抗的关系
收集临床NSCLC患者的病理标本及完整临床病历资料,并且设计随访问卷;同时采用免疫细胞化学检测NSCLC患者组织标本中APE1蛋白的表达,并分析其表达与NSCLCI临床分型、术后复发和含铂类药物化疗等的关系;
2、Ad5/F35-APE1 siRNA重组腺病毒增强肺腺癌顺铂化疗敏感性的实验研究
不同剂量顺铂和/或Ad5/F35-APE1 siRNA重组腺病毒处AA549细胞,MTT法检测其存活率的改变;TUNEL法检测A549细胞凋亡;Western blot检测APE1蛋白表达。
研究结果
l、APE1蛋白在NSCLC组织细胞的表达特点及NSCLC术后复发和含铂类药物抵抗的关系
正常肺组织及肺癌组织均有APE1表达,但正常肺组织以胞核表达为主,肺癌组织主要在胞浆表达;APE1的表达与患者的年龄、性别、肿瘤大小、有无淋巴结转移及组织类型无明显关系;APE1高表达组和APE1低表达组患者的1年生存率差异无统计学意义(χ~2=53.03,P＞0.05),而前者的2年和3年生存率低于后者,差异有统计学意义(P＜0.05)。24例铂类敏感组中,APE1强表达仅为2例,占8.33%,12例铂类耐药组APE1表达强阳性为10例,占83.33%,两两比较均有有统计学意义(χ~2=9.06,P＜0.01)。APE1基因检测有助于NSCLC疗效及预后的判断。
2、Ad5/F35-APEl siRNA重组腺病毒显著增强肺腺癌细胞顺铂化疗敏感性
顺铂呈剂量依赖性诱导A549细胞APE1蛋白表达增强,Ad5/F35-APE1 siRNA能显著抑制A549细胞APE1蛋白表达,且呈剂量依赖性。感染Ad5/F35-APE1 siRNA腺病毒比感染对照腺病毒的A549细胞对顺铂的敏感性明显增强,IC50分别为0.26μg/ml和1.54μg/ml。Ad5/F35-APE1 siRNA加强顺铂诱导的A549细胞凋亡。
结论
1、APE1胞浆异位表达可能与非小细胞肺癌的发生、侵袭和转移有关,同时APEl的表达强度与含铂类药物抵抗相关。APE1是非小细胞肺癌治疗潜在的分子靶点,具有重要的临床应用价值。
2、铂类化疗药物处理后肺癌细胞APE1蛋白表达水平明显升高,表明化疗药物在杀伤癌细胞的同时,癌细胞合成表达APE1增强DNA损伤修复,这可能是NSCLC癌细胞发生耐药的机制之一。
3、阻断NSCLC细胞APE1基因表达,可显著提高NSCLC细胞化疗敏感性,诱导NSCLC细胞发生凋亡。基于APEl基因的RNAi技术靶向增强NSCLC细胞化疗敏感性、诱导NSCLC细胞凋亡对提高NSCLC化疗疗效可能具有一定的临床应用前景。
Non-small-cell lung cancer(NSCLC) accounts for 80%of all lung cancer cases and is the leading cause of cancer mortality.The treatment of advanced NSCLC is based on the combination of platinum and one of the following agents:paclitaxel,docetaxel, gemcitabine or NVB.There are no significant differences in efficacy among these combinations suggesting that the outcome of platinum therapy on NSCLC have reached a plateau.Therefore,the biological mechanisms of cisplatin action need to be understood in order to overcome the treatment plateau on NSCLC.The molecular target of platinum action is the cellular DNA,which hampers DNA replication and transcription,resulting in cell death.Moreover,the development of resistance is a hurdle in the use of this drug.The molecular mechanisms that underlie this chemoresistance are largely unknown.Possible mechanisms of acquired resistance to platinum include reduced intracellular accumulation of platinum,enhanced drug inactivation by metallothionein and glutathione,increased repair activity of DNA damage,and formation of cisplatin-DNA adducts,et al.
DNA-repair systems,as the molecular basis of defending against environmental damage to cellular DNA,play an important role in protecting the genomic stabilization and integrity.However,an elevated DNA repair capacity in tumor cells leads to drug resistance and severely limits the efficacy of platinum.The human apurinic/apyrimidinic endonuclease (APE1),is abundant in human cells and accounts for nearly all of the abasic site cleavage activity observed in cellular extracts.In addition to its DNA repair functions,APE1 is also a multifunctional protein that participates in other crucial cellular processes,including the response to oxidative stress and regulation of transcription factors.The transcription factors are associated with chemoresistance.Therefore,targeting inhibition of APE1 display an emerging cancer therapeutic opportunity.In this study,we first investigate the expression of APE1 and its correlation with sensitivity of platinum chemotherapy in NSCLC patients. Then,we investigated the effect of adenoviral vector Ad5/F35 carrying human APE1 siRNA on the sensitivity of cisplatin in human NSCLC cells.
Objective
1.To research the expression of APE1 protein in NSCLC cells,analyze the relationship between APE1 expression and NSCLC clinical effect,and determinate that APE1 is an effective therapeutic target gene of NSCLC.
2.To investigate the perspective of clinical application of gene therapy targeting APE1 gene enhancing the sensitivity of NSCLC cells to DDP.
Materials and Methods
1.The expression of APE1 protein in NSCLC cells and its significance
NSCLC patients and their medical records were collected.The expression of APE1 in NSCLC patients and NSCLC cell lines(A549) were detected by immunocytochemical staining and Western blot,then analyzed relationship between expression of APE1 and clinical classification,clinical effect of NSCLC.
2.Study of Ad5/F35-APE1 siRNA enhancing sensitivity of human NSCLC cells to DDP
Cells were treated with DDP at various concentrations 48 hours after Ad5/F35-APE1 siRNA or Ad5/F35-EGFP was transfered into A549 cells,and the cellular proliferation capacity was observed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.Cell apoptosis was determined by TUNEL.The expression of human APE1 protein was detected by western blot analysis.
Results
1.The expression of APE1 protein in NSCLC cells and its significance
Protein expression of APE1 was noticed in all non-small cell lung carcinomas and normal lung tissue.The APE1 expression of normal lung tissue mainly located in the nucleus,while APE1 expression of non-small cell lung carcinomas tissue mainly located in cytoplasm.In addition,APE1 expression was not related with the age and gender of patients,tumor size,lymphonode metastasis,nor the pathologic classification.It was not statistical significance that the patient one-year survival rate between APE1- high expression group and the low expression group(x~2=53.03,P＞0.05),but in the 2 and 3-years survival rates of APE1 low expression higher than the high expression(P＜0.05).In the 12 examples platinum drug fast group,APE1- high expression was 10 examples(83.33%),and in the 24 examples platinum sensitiveness group,APE1- high expression was 2 examples (8.33%),they were also notable statistical slgnlficance(x~2=9.06,P＜0.01).To detect gene APE1 will be conduced to judge the therapeutic effect and prognostic of NSCLC.
2.Study of Ad5/F35-APE1 siRNA enhancing sensitivity of human NSCLC cells to DDP
The protein expression of APE1 in A549 cells was induced by DDP in a dosedependent manner.Infection of A549 cells with Ad5/F35-APE1 siRNA resulted in a dose-dependent suppression of APE1 protein in vitro.The result of MTT showed that Ad5/F35-APE1 siRNA enhanced sensitivity of A549 cells to DDP.The 50%inhibitory concentration(IC50) value for A549 cells pretreatmented with Ad5/F35-APE1 siRNA and Ad5/35-EGFP at 72 h after DDP treatment was 0.26μg/ml and 1.54μg/ml,respectively. Ad5/F35-APE1 siRNA also increased cell apoptosis induction by DDP.
Conclusion
1.The shifts of APE1 from nucleus to cytoplasm might play a pivotal role in the carcinogenesis,progression and metastasis of NSCLC.High expression of APE1 protein may indicate poor prognosis,and correlated with the DDP drug resistance.It suggests that APE1 is a potential molecular therapeutic target of NSCLC.
2.The protein expression of APE1 in NSCLC cells was induced by DDP in a dose-dependent manner,which suggests that elevated DNA repair capacity may partially contribute to the resistance of DDP in NSCLC cells.
3.Inhibiting APE1 expression could improve sensitivityof DDP remarkably and induce apoptosis in NSCLC cells.Therefore,gene therapy targeting APE1 gene shows a promising approach in enhancing the sensitivity of NSCLC to chemotherapy.
DNA损伤修复系统作为机体抵抗各种损伤的分子基础,对于维护基因组的稳定与完整起着至关重要的作用,然而对于通过损伤DNA杀死癌细胞的铂类化疗,这一过程无疑削弱了治疗效果。因此,DNA修复能力增强可能是铂类耐药的主要机制之一。DNA损伤修复基因脱嘌呤/脱嘧啶核酸内切酶(apurinic/apyrimidinic endonuclease,AP,1具有核酸内切酶及氧化还原双功能,是细胞放射性损伤和基因毒性药物烷化剂致伤的重要修复因子,并通过氧化还原机制调节多种转录因子及下游靶基因表达,而这些转录因子与肿瘤放化疗抵抗密切相关。因此本研究检测NSCLC患者APE1蛋白的表达特点,分析APEl与NSCLC术后复发和铂类化疗敏感性的关系,并用Ad5/F35-APE1 siRNA重组腺病毒载体特异性“敲除”APE1基因表达,研究其体外对NSCLC细胞顺铂化疗敏感性的影响。
研究目的:
1、研究APE1蛋白在非小细胞肺癌组织细胞中表达情况,分析APE1表达特点与NSCLC术后复发和铂类抵抗的关系,以确证APE1为NSCLC治疗有效的靶基因。
2、探讨诱导APE1基因沉默对NSCLC含铂类化疗的增敏作用。
研究内容和方法:
1、APE1蛋白在NSCLC细胞中的表达特点及NSCLC术后复发和含铂类药物抵抗的关系
收集临床NSCLC患者的病理标本及完整临床病历资料,并且设计随访问卷;同时采用免疫细胞化学检测NSCLC患者组织标本中APE1蛋白的表达,并分析其表达与NSCLCI临床分型、术后复发和含铂类药物化疗等的关系;
2、Ad5/F35-APE1 siRNA重组腺病毒增强肺腺癌顺铂化疗敏感性的实验研究
不同剂量顺铂和/或Ad5/F35-APE1 siRNA重组腺病毒处AA549细胞,MTT法检测其存活率的改变;TUNEL法检测A549细胞凋亡;Western blot检测APE1蛋白表达。
研究结果
l、APE1蛋白在NSCLC组织细胞的表达特点及NSCLC术后复发和含铂类药物抵抗的关系
正常肺组织及肺癌组织均有APE1表达,但正常肺组织以胞核表达为主,肺癌组织主要在胞浆表达;APE1的表达与患者的年龄、性别、肿瘤大小、有无淋巴结转移及组织类型无明显关系;APE1高表达组和APE1低表达组患者的1年生存率差异无统计学意义(χ~2=53.03,P＞0.05),而前者的2年和3年生存率低于后者,差异有统计学意义(P＜0.05)。24例铂类敏感组中,APE1强表达仅为2例,占8.33%,12例铂类耐药组APE1表达强阳性为10例,占83.33%,两两比较均有有统计学意义(χ~2=9.06,P＜0.01)。APE1基因检测有助于NSCLC疗效及预后的判断。
2、Ad5/F35-APEl siRNA重组腺病毒显著增强肺腺癌细胞顺铂化疗敏感性
顺铂呈剂量依赖性诱导A549细胞APE1蛋白表达增强,Ad5/F35-APE1 siRNA能显著抑制A549细胞APE1蛋白表达,且呈剂量依赖性。感染Ad5/F35-APE1 siRNA腺病毒比感染对照腺病毒的A549细胞对顺铂的敏感性明显增强,IC50分别为0.26μg/ml和1.54μg/ml。Ad5/F35-APE1 siRNA加强顺铂诱导的A549细胞凋亡。
结论
1、APE1胞浆异位表达可能与非小细胞肺癌的发生、侵袭和转移有关,同时APEl的表达强度与含铂类药物抵抗相关。APE1是非小细胞肺癌治疗潜在的分子靶点,具有重要的临床应用价值。
2、铂类化疗药物处理后肺癌细胞APE1蛋白表达水平明显升高,表明化疗药物在杀伤癌细胞的同时,癌细胞合成表达APE1增强DNA损伤修复,这可能是NSCLC癌细胞发生耐药的机制之一。
3、阻断NSCLC细胞APE1基因表达,可显著提高NSCLC细胞化疗敏感性,诱导NSCLC细胞发生凋亡。基于APEl基因的RNAi技术靶向增强NSCLC细胞化疗敏感性、诱导NSCLC细胞凋亡对提高NSCLC化疗疗效可能具有一定的临床应用前景。
Non-small-cell lung cancer(NSCLC) accounts for 80%of all lung cancer cases and is the leading cause of cancer mortality.The treatment of advanced NSCLC is based on the combination of platinum and one of the following agents:paclitaxel,docetaxel, gemcitabine or NVB.There are no significant differences in efficacy among these combinations suggesting that the outcome of platinum therapy on NSCLC have reached a plateau.Therefore,the biological mechanisms of cisplatin action need to be understood in order to overcome the treatment plateau on NSCLC.The molecular target of platinum action is the cellular DNA,which hampers DNA replication and transcription,resulting in cell death.Moreover,the development of resistance is a hurdle in the use of this drug.The molecular mechanisms that underlie this chemoresistance are largely unknown.Possible mechanisms of acquired resistance to platinum include reduced intracellular accumulation of platinum,enhanced drug inactivation by metallothionein and glutathione,increased repair activity of DNA damage,and formation of cisplatin-DNA adducts,et al.
DNA-repair systems,as the molecular basis of defending against environmental damage to cellular DNA,play an important role in protecting the genomic stabilization and integrity.However,an elevated DNA repair capacity in tumor cells leads to drug resistance and severely limits the efficacy of platinum.The human apurinic/apyrimidinic endonuclease (APE1),is abundant in human cells and accounts for nearly all of the abasic site cleavage activity observed in cellular extracts.In addition to its DNA repair functions,APE1 is also a multifunctional protein that participates in other crucial cellular processes,including the response to oxidative stress and regulation of transcription factors.The transcription factors are associated with chemoresistance.Therefore,targeting inhibition of APE1 display an emerging cancer therapeutic opportunity.In this study,we first investigate the expression of APE1 and its correlation with sensitivity of platinum chemotherapy in NSCLC patients. Then,we investigated the effect of adenoviral vector Ad5/F35 carrying human APE1 siRNA on the sensitivity of cisplatin in human NSCLC cells.
Objective
1.To research the expression of APE1 protein in NSCLC cells,analyze the relationship between APE1 expression and NSCLC clinical effect,and determinate that APE1 is an effective therapeutic target gene of NSCLC.
2.To investigate the perspective of clinical application of gene therapy targeting APE1 gene enhancing the sensitivity of NSCLC cells to DDP.
Materials and Methods
1.The expression of APE1 protein in NSCLC cells and its significance
NSCLC patients and their medical records were collected.The expression of APE1 in NSCLC patients and NSCLC cell lines(A549) were detected by immunocytochemical staining and Western blot,then analyzed relationship between expression of APE1 and clinical classification,clinical effect of NSCLC.
2.Study of Ad5/F35-APE1 siRNA enhancing sensitivity of human NSCLC cells to DDP
Cells were treated with DDP at various concentrations 48 hours after Ad5/F35-APE1 siRNA or Ad5/F35-EGFP was transfered into A549 cells,and the cellular proliferation capacity was observed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.Cell apoptosis was determined by TUNEL.The expression of human APE1 protein was detected by western blot analysis.
Results
1.The expression of APE1 protein in NSCLC cells and its significance
Protein expression of APE1 was noticed in all non-small cell lung carcinomas and normal lung tissue.The APE1 expression of normal lung tissue mainly located in the nucleus,while APE1 expression of non-small cell lung carcinomas tissue mainly located in cytoplasm.In addition,APE1 expression was not related with the age and gender of patients,tumor size,lymphonode metastasis,nor the pathologic classification.It was not statistical significance that the patient one-year survival rate between APE1- high expression group and the low expression group(x~2=53.03,P＞0.05),but in the 2 and 3-years survival rates of APE1 low expression higher than the high expression(P＜0.05).In the 12 examples platinum drug fast group,APE1- high expression was 10 examples(83.33%),and in the 24 examples platinum sensitiveness group,APE1- high expression was 2 examples (8.33%),they were also notable statistical slgnlficance(x~2=9.06,P＜0.01).To detect gene APE1 will be conduced to judge the therapeutic effect and prognostic of NSCLC.
2.Study of Ad5/F35-APE1 siRNA enhancing sensitivity of human NSCLC cells to DDP
The protein expression of APE1 in A549 cells was induced by DDP in a dosedependent manner.Infection of A549 cells with Ad5/F35-APE1 siRNA resulted in a dose-dependent suppression of APE1 protein in vitro.The result of MTT showed that Ad5/F35-APE1 siRNA enhanced sensitivity of A549 cells to DDP.The 50%inhibitory concentration(IC50) value for A549 cells pretreatmented with Ad5/F35-APE1 siRNA and Ad5/35-EGFP at 72 h after DDP treatment was 0.26μg/ml and 1.54μg/ml,respectively. Ad5/F35-APE1 siRNA also increased cell apoptosis induction by DDP.
Conclusion
1.The shifts of APE1 from nucleus to cytoplasm might play a pivotal role in the carcinogenesis,progression and metastasis of NSCLC.High expression of APE1 protein may indicate poor prognosis,and correlated with the DDP drug resistance.It suggests that APE1 is a potential molecular therapeutic target of NSCLC.
2.The protein expression of APE1 in NSCLC cells was induced by DDP in a dose-dependent manner,which suggests that elevated DNA repair capacity may partially contribute to the resistance of DDP in NSCLC cells.
3.Inhibiting APE1 expression could improve sensitivityof DDP remarkably and induce apoptosis in NSCLC cells.Therefore,gene therapy targeting APE1 gene shows a promising approach in enhancing the sensitivity of NSCLC to chemotherapy.
引文
1. Evans AR, Limp-Foster M,Kelley MR. Going Apel over ref-1. Mutat Res. 2000; 461: 104.
2. Moore DH, Michael H, Tritt R, et al. Alterationsnin the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. ClinCancer Res. 2000; 6 (2):607.
3. Kelley MR, Cheng L, Foster R, et al. Elevated and altered expression of the multifunctional DNA base excision repair and redox enzyme Apel/ref-1 in prostate cancer. Clin Cancer Res. 2001;7 (4):828
4. Puglisi F, Barbone F, Tell G, et al. Prognostic role of Ape/Ref-1 subcellar expression in stage I -III breast carcinomas. Oncol Rep. 2002;9 (1):11.
5. Dong Wang,Meihua Luo, Mark Kelley. Human apurinic endonucleasa 1 (APE1) expression and prognostic significance in osteosarcoma: Enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004;3 (6): 682.
6. Mallery DL, Tanganelli B, Colella S, et al. Molecular analysis of mutations in the CSB (ERCC6) gene in patients with Cockayne syndrome. Am J Hum Genet. 1998;62(1):80.
7. Shen H,Sturgis EM,Khan SG,et al. An intronic poly (AT) polymorphism of the DNA repair gene XPC and risk of squamous cell carcinoma of the head and neck: a case-control study. Cancer Res. 2001; 15;61(8):3324.
8. Xing DY, Tan W,Song N,et al. Ser326Cys polymorphism in hOGG1 gene and risk of esophageal cancer in a Chinese population. Int J Cancer. 2001; 95 (3): 142.
9. Winsey SL, Haldar NA, Marsh HP, et al. A variant within the DNA repair gene XRCC3 is associated with the development of melanoma skin cancer. Cancer Res. 2000;60(20):5614.
10. Peltomaki P, Vasen HF. Mutations predisposing to hereditary nonpolyposis colorectal cancer: database and results of a collaborative study. The International Collaborative Group on Hereditary Nonpolyposis Colorectal Cancer. Gastroenterology. 1997; 113 (4): 1150.
11. Ramana CV, Boldogh I, Izumi T, et al. Activation of apurinic/apyrimidinic endonuclease in human cells by reactive oxygen species and its correlation with their adaptive response to genotoxicity of free radicals. Proc Natl Acad Sci U S A. 1998; 95 (9):5064.
12. Xanthoudakis S, Curran T. Identification and characterization of APE/Ref-1, a nuclear protein that facilitates AP-1 DNA-binding activity. EMBOJ, 1992; 11(2):662.
13. Okazaki T, Chung U, Nishita T, et al. A redox factor protein, ref-1, is involved in negative gene regulation by extra cellular calcium. J Biol Chem. 1994; 269 (45):27860.
14. Ueno M, Masutant H, Arai RJ, et al. Thioredoxix independent redox regulation of p53 mediated p21 activation. J Biol Chem. 1999; 274(50):35813.
15. Kakolyris S, Giatrmanolaki A, Kackourakis M, et al. Nuclear location of human AP endonuclease 1(APE1/Ref-1) associates with prognosis nearly operable non-small cell lung cancer(NSCLC). J Pathol. 1999;189(3):356.
16. Xanthoudakis S, Miao G, Wang F, et al. Redox activation of Fos-Jun DNA binding activity is mediated by a DNA repair enzyme. EMBOJ. 1992; 11(9):3330.
17. Takiguchi Y, Chen DJ. Genomic structure of the mouse apurinic/apyrimidinic endonuclease gene. Mamm Genome.1994;5(11):719.
18. Seki S, Ikeda S, Watanabe S, et al. A mouse DNA repair enzyme (APEX nuclease) having exonuclease and apurinic/apyrimidinic endonuclease activities: purification and characterization. Biochim Biophys Acta, 1991; 1079(1):61.
19. Kakolyrix S, Kakoamanis L, Engels K, et al. Human AP endonucease 1(HAP 1) protein expression in breast cancer or relate with lymphnode status and angiogenesis. Br J Cancer, 1998;77(7): 1171.
20. Kawase M, Fujimura M, Morita-Fujimura Y, et al. Reduction of apurinic/apyrimidnic endonucease expression after transient global cerebral ischemia in rats: implication of the failure of DNA repair in neuronal apoptosis. Stroke, 1999; 30 (2):448.
21. Gillardon F, Bottiger B, Hossmann KA. Expression of nuclear redox factor APE/Ref-1 in the rat hippocampus following global is chemia induced by cardiac arrest. Brain Res Mol Brain Res. 1997; 52(2): 199.
22. Evans AR, Limp Foster M,Kelley MR. Going APE over ref-1[J]. Mutat Res, 2000, 461(2):89.
23. Angkeow P, Deshpande SS, Qi B, et al . Redox factor-1: an extranuclearrole in the regulation of endothelial oxidative stress and apoptosis [J].Cell Death Differ, 2002, 9 (7) :720.
24. Puglisi F, Barbone F, Tell G, et al. Prognostic role of Ape1/Ref-1 subcellar expression in stage I - III breast carcinomas [J ] . Oncol Rep, 2002, 9 (1):11.
25. Tanner B, Grimme S, Schiffer I, et al. Nuclear expression of apurinic/ apyrimidinic endonuclease increases with progression of ovarian carcinomas [J]. Gynecologic Oncology, 2004, 92 (2):568 -577.
26. Silber JR, Bobola MS, Blank A, et al. The apurinic/apyrimidinic endonuclease activity of Apel/Ref-1 contributes to human glioma cell resistance to alkylating agents and is elevated by oxidative stress. Clin Cancer Res, 2002; 8: 3014.
27. Tai Y T,Teoh G,Lin B,et al . Ku86 variant expression and function in multiple myeloma cells is associated with increased sentivity to DNA damage [J]. J Immunol, 2000,165(11): 6347-6355.
28. Efferth T,Fabry U,Osieka R.Interleukin26 affects melphalan2induced DNA damage and repair in human multiple myeloma cells [J]. Anticancer Res, 2002, 22 (1A):231-234.
29. Koukourakis MI, Giatromanolaki A, Kakolyris S,et al. Nuclear expression of human apurinic/apyrimidinic endonuclease (HAP1/Ref-1) in head-and-neck cancer is associated with resistance to chemoradiotherapy and poor outcome. Int J Radiat Oncol Biol Phys,2001,50(1):34.
30. Moore DH, Michael H, Tritt R, et al. Alterations in the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. Clin Cancer Res, 2000, 6 (2): 606.
31. Wang D, Luo M, Kelley MR. Human apurinic endonuclease 1 (APE1) expression and prognostic significance in osteosarcoma: enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004, 3(6): 683.
32. Ono Y, Furuta T, Ohmoto T, et al. Stable expression in rat glioma cells of sense and antisense nucleic acids to a human multifunctional DNA repair enzyme APEX nuclease. Mutat Res, 1994, 315(1):61.
33. McNeill DR, Wilson DM 3rd. A dominant-negative form of the major human abasic endonuclease enhances cellular sensitivity to laboratory and clinical DNA-damaging agents. Mol Cancer Res, 2007, 5(1):70.
34. Yang ZZ, Chen XH, Wang D. Experimental study enhancing the chemosensitivity of multiple myeloma to melphalan by using a tissue-specific APE1-silencing RNA expression vector.Clin Lymphoma Myeloma. 2007, 7(4):301.
35. Angkeow P, Deshpande SS, Qi B, et al. Redox factor-1: an extranuclearrole in the regulation of endothelial oxidative stress and apoptosis [J] .Cell Death Differ, 2002, 9 (7): 721.
36. Hall JL, Wang X, Van Adamson V, et al. Overexpression of Ref-1 inhibits hypoxia and tumor necrosis factor induced endothelial cell apoptosis through nuclear factor kappab independent and dependent pathways [J]. Circ Res, 2001, 88 (12): 1250.
37. Evans AR, Limp Foster M, Kelley MR. Going APE over ref-1 [J]. Mutat Res, 2000, 461 (2): 100.
38. Gaiddon C, Moorthy NC, Prives C. Ref-1 regulates the transactivation and proapoptotic functions of p53 in vivo [J]. EMBOJ, 1999, 18 (20):5617.
39. Shaulian E, Karin M. AP21 as a regulator of cell life and death [J]. Nat Cell Biol, 2002, 4 (5): 134.
40. Wang N, Stemerman MB. Ref-1 and transcriptional control of endothelial apoptosis[J]. Circ Res, 2001, 88(12): 1223-1225.
41. Wang D, Luo M, Kelley MR. Human apurinic endonuclease 1 (APE1) expression and prognostic significance in osteosarcoma: enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004, 3(6): 679-686.
42. Koukourakis MI, Giatromanolaki A, Kakolyris S, et al. Nuclear expression of human apurinic/apyrimidinic endonuclease (HAP1/Ref-1) in head-and-neck cancer is associated with resistance to chemoradiotherapy and poor outcome. Int J Radiat Oncol Biol Phys, 2001, 50(1):27-36.
1.Barzilay G,Hickson ID.Structure and function of apurinic/apyrimidinic endonucleases.Bioessays,1995,17:713-719.
2.Barzilay G,Mol CD,Robson CN,et al.Identification of critical active site residues in the multifunctional human DNA repair enzyme HAP1.Nat Struct Biol,1995,2:561-565.
3.Barzilay G,Walker LJ,Robson CN,etal.Site directed mutagenesis of the human DNA repai enzyme HAP1:identification of residues important for AP endonuclease and Rnase H activity.Nucleic Acids Res,1995,23:1544-1550.
4.Moore DH,Michael H,Tritt R,et al.Alterations in the expression of the DNA repair redox enzyme APE1/ref-1 in epithelial ovarian cancers[J].Clin Cancer Res,2000,6(2):602-609.
5.Tell G,Pellizzari L,Pucillo C,et al.TSH controls Ref21 nuclear translo2cation in thyroid cells[J].J Mol Endocrinol,2000,24(3):383-390.
6.Kelley MR,Cheng L,Foster R,et al.Elevated and altered expression of the multifunctional DNA base excision repair and redox enzyme Ape1Pref21 in prostate cancer [J]. Clin Cancer Res, 2001, 7(4):824-830.
7. Puglisi F, Barbone F, Tell G, et al. Prognostic role of Ape1/Ref-1 subcellar expression in stage I - III breast carcinomas [J]. Oncol Rep, 2002, 9(1) : 11 — 17.
8. Kakolyris S, Giatromanolaki A, Koukourakis M, et al. Nuclear location ofhuman AP endonuclease1 (APE1/Ref-1) associates with prognosis in earlyoperable non2small cell lung cancer (NSCLC) [J] . J Pathol, 1999, 189 (3) :351-357.
9. Hadi MZ, Coleman MA, Fidelis K, et al. Functional characterization of Ape1 variants identified in the human population [J]. Nucleic Acids Res, 2000, 28 (20):3871-3879.
10. Shinmura K, Yamaguchi S, Saitoh T, et al. Somatic mutations and single nucleotide polymorphisms of base excision repair genes involved in the repair of hydroxyguanine in damaged DNA [J] . Cancer Lett, 2001, 166(1):65-69.
11. Pieretti M, Khattar NH, Smith SA. Common polymorphisms and somaticmutations in human base excision repair genes in ovarian and endometrial cancers [J]. Mutat Res, 2001, 432 (324) :53-59.
12. Hu JJ, Smith TR, Miller MS, et al. Amino acid substitution variants of APE1 and XRCC1 genes associated with ionizing radiation sensitivity[J ] . Carcinogenesis, 2001, 22 (6) :917-922.
13. Misra RR, Ratnasinghe D, Tangrea JA, et al. Polymorphisms in the DNA repair genes XPD, XRCC1, XRCC3, and APE1/ref-1, and the risk of lung cancer among male smokers in Finland [J]. Cancer Lett, 2003, 191(2): 171-178.
14. Angkeow P, Deshpande SS, Qi B, et al. Redox factor-1: an extranuclearrole in the regulation of endothelial oxidative stress and apoptosis [J] .Cell Death Differ, 2002, 9 (7): 717-725.
15. Hall JL, Wang X,Van Adamson V, et al. Overexpression of Ref-1 inhibits hypoxia and tumor necrosis factor induced endothelial cell apoptosis through nuclear factor kappab independent and dependent pathways [J]. Circ Res, 2001, 88 (12) : 1247-1253.
16. Evans AR, Limp Foster M, Kelley MR. Going APE over ref-1 [J] . Mutat Res, 2000,461 (2):83-108.
17. Gaiddon C, Moorthy NC, Prives C. Ref-1 regulates the transactivation and proapoptotic functions of p53 in vivo [J]. EMBOJ, 1999, 18(20) :5609-5621.
18. Shaulian E, Karin M. APE1 as a regulator of cell life and death [J]. Nat Cell Biol, 2002, 4(5) : E131-136.
19. Wang N, Stemerman MB. Ref-1 and transcriptional control of endothelial apoptosis[J]. Circ Res, 2001, 88(12): 1223-1225.
20. Wang D, Luo M, Kelley MR. Human apurinic endonuclease 1 (APE1) expression and prognostic significance in osteosarcoma: enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004, 3(6): 679-686.
21. Koukourakis MI, Giatromanolaki A, Kakolyris S, et al. Nuclear expression of human apurinic/apyrimidinic endonuclease (HAP1/Ref-1) in head-and-neck cancer is associated with resistance to chemoradiotherapy and poor outcome. Int J Radiat Oncol Biol Phys, 2001, 50(1):27-36.
22. Moore DH, Michael H, Tritt R, et al. Alterations in the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. Clin Cancer Res, 2000, 6(2):602-609.
23. Reuven Agami. RNAi and related mechanisms and their potential use for therapy. Current Opinion in Chemical Biology, 2002, 6: 829-834.
24. Elbashir SM, Harborth J, Lendeckel W, et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature, 2001, 411(6836):494-498
25. Ghosh SS, Gopinath P, Ramesh A. Adenoviral vectors: a promising tool for gene therapy. Appl Biochem Biotechnol, 2006, 133(1): 9-29.
2. Moore DH, Michael H, Tritt R, et al. Alterationsnin the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. ClinCancer Res. 2000; 6 (2):607.
3. Kelley MR, Cheng L, Foster R, et al. Elevated and altered expression of the multifunctional DNA base excision repair and redox enzyme Apel/ref-1 in prostate cancer. Clin Cancer Res. 2001;7 (4):828
4. Puglisi F, Barbone F, Tell G, et al. Prognostic role of Ape/Ref-1 subcellar expression in stage I -III breast carcinomas. Oncol Rep. 2002;9 (1):11.
5. Dong Wang,Meihua Luo, Mark Kelley. Human apurinic endonucleasa 1 (APE1) expression and prognostic significance in osteosarcoma: Enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004;3 (6): 682.
6. Mallery DL, Tanganelli B, Colella S, et al. Molecular analysis of mutations in the CSB (ERCC6) gene in patients with Cockayne syndrome. Am J Hum Genet. 1998;62(1):80.
7. Shen H,Sturgis EM,Khan SG,et al. An intronic poly (AT) polymorphism of the DNA repair gene XPC and risk of squamous cell carcinoma of the head and neck: a case-control study. Cancer Res. 2001; 15;61(8):3324.
8. Xing DY, Tan W,Song N,et al. Ser326Cys polymorphism in hOGG1 gene and risk of esophageal cancer in a Chinese population. Int J Cancer. 2001; 95 (3): 142.
9. Winsey SL, Haldar NA, Marsh HP, et al. A variant within the DNA repair gene XRCC3 is associated with the development of melanoma skin cancer. Cancer Res. 2000;60(20):5614.
10. Peltomaki P, Vasen HF. Mutations predisposing to hereditary nonpolyposis colorectal cancer: database and results of a collaborative study. The International Collaborative Group on Hereditary Nonpolyposis Colorectal Cancer. Gastroenterology. 1997; 113 (4): 1150.
11. Ramana CV, Boldogh I, Izumi T, et al. Activation of apurinic/apyrimidinic endonuclease in human cells by reactive oxygen species and its correlation with their adaptive response to genotoxicity of free radicals. Proc Natl Acad Sci U S A. 1998; 95 (9):5064.
12. Xanthoudakis S, Curran T. Identification and characterization of APE/Ref-1, a nuclear protein that facilitates AP-1 DNA-binding activity. EMBOJ, 1992; 11(2):662.
13. Okazaki T, Chung U, Nishita T, et al. A redox factor protein, ref-1, is involved in negative gene regulation by extra cellular calcium. J Biol Chem. 1994; 269 (45):27860.
14. Ueno M, Masutant H, Arai RJ, et al. Thioredoxix independent redox regulation of p53 mediated p21 activation. J Biol Chem. 1999; 274(50):35813.
15. Kakolyris S, Giatrmanolaki A, Kackourakis M, et al. Nuclear location of human AP endonuclease 1(APE1/Ref-1) associates with prognosis nearly operable non-small cell lung cancer(NSCLC). J Pathol. 1999;189(3):356.
16. Xanthoudakis S, Miao G, Wang F, et al. Redox activation of Fos-Jun DNA binding activity is mediated by a DNA repair enzyme. EMBOJ. 1992; 11(9):3330.
17. Takiguchi Y, Chen DJ. Genomic structure of the mouse apurinic/apyrimidinic endonuclease gene. Mamm Genome.1994;5(11):719.
18. Seki S, Ikeda S, Watanabe S, et al. A mouse DNA repair enzyme (APEX nuclease) having exonuclease and apurinic/apyrimidinic endonuclease activities: purification and characterization. Biochim Biophys Acta, 1991; 1079(1):61.
19. Kakolyrix S, Kakoamanis L, Engels K, et al. Human AP endonucease 1(HAP 1) protein expression in breast cancer or relate with lymphnode status and angiogenesis. Br J Cancer, 1998;77(7): 1171.
20. Kawase M, Fujimura M, Morita-Fujimura Y, et al. Reduction of apurinic/apyrimidnic endonucease expression after transient global cerebral ischemia in rats: implication of the failure of DNA repair in neuronal apoptosis. Stroke, 1999; 30 (2):448.
21. Gillardon F, Bottiger B, Hossmann KA. Expression of nuclear redox factor APE/Ref-1 in the rat hippocampus following global is chemia induced by cardiac arrest. Brain Res Mol Brain Res. 1997; 52(2): 199.
22. Evans AR, Limp Foster M,Kelley MR. Going APE over ref-1[J]. Mutat Res, 2000, 461(2):89.
23. Angkeow P, Deshpande SS, Qi B, et al . Redox factor-1: an extranuclearrole in the regulation of endothelial oxidative stress and apoptosis [J].Cell Death Differ, 2002, 9 (7) :720.
24. Puglisi F, Barbone F, Tell G, et al. Prognostic role of Ape1/Ref-1 subcellar expression in stage I - III breast carcinomas [J ] . Oncol Rep, 2002, 9 (1):11.
25. Tanner B, Grimme S, Schiffer I, et al. Nuclear expression of apurinic/ apyrimidinic endonuclease increases with progression of ovarian carcinomas [J]. Gynecologic Oncology, 2004, 92 (2):568 -577.
26. Silber JR, Bobola MS, Blank A, et al. The apurinic/apyrimidinic endonuclease activity of Apel/Ref-1 contributes to human glioma cell resistance to alkylating agents and is elevated by oxidative stress. Clin Cancer Res, 2002; 8: 3014.
27. Tai Y T,Teoh G,Lin B,et al . Ku86 variant expression and function in multiple myeloma cells is associated with increased sentivity to DNA damage [J]. J Immunol, 2000,165(11): 6347-6355.
28. Efferth T,Fabry U,Osieka R.Interleukin26 affects melphalan2induced DNA damage and repair in human multiple myeloma cells [J]. Anticancer Res, 2002, 22 (1A):231-234.
29. Koukourakis MI, Giatromanolaki A, Kakolyris S,et al. Nuclear expression of human apurinic/apyrimidinic endonuclease (HAP1/Ref-1) in head-and-neck cancer is associated with resistance to chemoradiotherapy and poor outcome. Int J Radiat Oncol Biol Phys,2001,50(1):34.
30. Moore DH, Michael H, Tritt R, et al. Alterations in the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. Clin Cancer Res, 2000, 6 (2): 606.
31. Wang D, Luo M, Kelley MR. Human apurinic endonuclease 1 (APE1) expression and prognostic significance in osteosarcoma: enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004, 3(6): 683.
32. Ono Y, Furuta T, Ohmoto T, et al. Stable expression in rat glioma cells of sense and antisense nucleic acids to a human multifunctional DNA repair enzyme APEX nuclease. Mutat Res, 1994, 315(1):61.
33. McNeill DR, Wilson DM 3rd. A dominant-negative form of the major human abasic endonuclease enhances cellular sensitivity to laboratory and clinical DNA-damaging agents. Mol Cancer Res, 2007, 5(1):70.
34. Yang ZZ, Chen XH, Wang D. Experimental study enhancing the chemosensitivity of multiple myeloma to melphalan by using a tissue-specific APE1-silencing RNA expression vector.Clin Lymphoma Myeloma. 2007, 7(4):301.
35. Angkeow P, Deshpande SS, Qi B, et al. Redox factor-1: an extranuclearrole in the regulation of endothelial oxidative stress and apoptosis [J] .Cell Death Differ, 2002, 9 (7): 721.
36. Hall JL, Wang X, Van Adamson V, et al. Overexpression of Ref-1 inhibits hypoxia and tumor necrosis factor induced endothelial cell apoptosis through nuclear factor kappab independent and dependent pathways [J]. Circ Res, 2001, 88 (12): 1250.
37. Evans AR, Limp Foster M, Kelley MR. Going APE over ref-1 [J]. Mutat Res, 2000, 461 (2): 100.
38. Gaiddon C, Moorthy NC, Prives C. Ref-1 regulates the transactivation and proapoptotic functions of p53 in vivo [J]. EMBOJ, 1999, 18 (20):5617.
39. Shaulian E, Karin M. AP21 as a regulator of cell life and death [J]. Nat Cell Biol, 2002, 4 (5): 134.
40. Wang N, Stemerman MB. Ref-1 and transcriptional control of endothelial apoptosis[J]. Circ Res, 2001, 88(12): 1223-1225.
41. Wang D, Luo M, Kelley MR. Human apurinic endonuclease 1 (APE1) expression and prognostic significance in osteosarcoma: enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004, 3(6): 679-686.
42. Koukourakis MI, Giatromanolaki A, Kakolyris S, et al. Nuclear expression of human apurinic/apyrimidinic endonuclease (HAP1/Ref-1) in head-and-neck cancer is associated with resistance to chemoradiotherapy and poor outcome. Int J Radiat Oncol Biol Phys, 2001, 50(1):27-36.
1.Barzilay G,Hickson ID.Structure and function of apurinic/apyrimidinic endonucleases.Bioessays,1995,17:713-719.
2.Barzilay G,Mol CD,Robson CN,et al.Identification of critical active site residues in the multifunctional human DNA repair enzyme HAP1.Nat Struct Biol,1995,2:561-565.
3.Barzilay G,Walker LJ,Robson CN,etal.Site directed mutagenesis of the human DNA repai enzyme HAP1:identification of residues important for AP endonuclease and Rnase H activity.Nucleic Acids Res,1995,23:1544-1550.
4.Moore DH,Michael H,Tritt R,et al.Alterations in the expression of the DNA repair redox enzyme APE1/ref-1 in epithelial ovarian cancers[J].Clin Cancer Res,2000,6(2):602-609.
5.Tell G,Pellizzari L,Pucillo C,et al.TSH controls Ref21 nuclear translo2cation in thyroid cells[J].J Mol Endocrinol,2000,24(3):383-390.
6.Kelley MR,Cheng L,Foster R,et al.Elevated and altered expression of the multifunctional DNA base excision repair and redox enzyme Ape1Pref21 in prostate cancer [J]. Clin Cancer Res, 2001, 7(4):824-830.
7. Puglisi F, Barbone F, Tell G, et al. Prognostic role of Ape1/Ref-1 subcellar expression in stage I - III breast carcinomas [J]. Oncol Rep, 2002, 9(1) : 11 — 17.
8. Kakolyris S, Giatromanolaki A, Koukourakis M, et al. Nuclear location ofhuman AP endonuclease1 (APE1/Ref-1) associates with prognosis in earlyoperable non2small cell lung cancer (NSCLC) [J] . J Pathol, 1999, 189 (3) :351-357.
9. Hadi MZ, Coleman MA, Fidelis K, et al. Functional characterization of Ape1 variants identified in the human population [J]. Nucleic Acids Res, 2000, 28 (20):3871-3879.
10. Shinmura K, Yamaguchi S, Saitoh T, et al. Somatic mutations and single nucleotide polymorphisms of base excision repair genes involved in the repair of hydroxyguanine in damaged DNA [J] . Cancer Lett, 2001, 166(1):65-69.
11. Pieretti M, Khattar NH, Smith SA. Common polymorphisms and somaticmutations in human base excision repair genes in ovarian and endometrial cancers [J]. Mutat Res, 2001, 432 (324) :53-59.
12. Hu JJ, Smith TR, Miller MS, et al. Amino acid substitution variants of APE1 and XRCC1 genes associated with ionizing radiation sensitivity[J ] . Carcinogenesis, 2001, 22 (6) :917-922.
13. Misra RR, Ratnasinghe D, Tangrea JA, et al. Polymorphisms in the DNA repair genes XPD, XRCC1, XRCC3, and APE1/ref-1, and the risk of lung cancer among male smokers in Finland [J]. Cancer Lett, 2003, 191(2): 171-178.
14. Angkeow P, Deshpande SS, Qi B, et al. Redox factor-1: an extranuclearrole in the regulation of endothelial oxidative stress and apoptosis [J] .Cell Death Differ, 2002, 9 (7): 717-725.
15. Hall JL, Wang X,Van Adamson V, et al. Overexpression of Ref-1 inhibits hypoxia and tumor necrosis factor induced endothelial cell apoptosis through nuclear factor kappab independent and dependent pathways [J]. Circ Res, 2001, 88 (12) : 1247-1253.
16. Evans AR, Limp Foster M, Kelley MR. Going APE over ref-1 [J] . Mutat Res, 2000,461 (2):83-108.
17. Gaiddon C, Moorthy NC, Prives C. Ref-1 regulates the transactivation and proapoptotic functions of p53 in vivo [J]. EMBOJ, 1999, 18(20) :5609-5621.
18. Shaulian E, Karin M. APE1 as a regulator of cell life and death [J]. Nat Cell Biol, 2002, 4(5) : E131-136.
19. Wang N, Stemerman MB. Ref-1 and transcriptional control of endothelial apoptosis[J]. Circ Res, 2001, 88(12): 1223-1225.
20. Wang D, Luo M, Kelley MR. Human apurinic endonuclease 1 (APE1) expression and prognostic significance in osteosarcoma: enhanced sensitivity of osteosarcoma to DNA damaging agents using silencing RNA APE1 expression inhibition. Mol Cancer Ther, 2004, 3(6): 679-686.
21. Koukourakis MI, Giatromanolaki A, Kakolyris S, et al. Nuclear expression of human apurinic/apyrimidinic endonuclease (HAP1/Ref-1) in head-and-neck cancer is associated with resistance to chemoradiotherapy and poor outcome. Int J Radiat Oncol Biol Phys, 2001, 50(1):27-36.
22. Moore DH, Michael H, Tritt R, et al. Alterations in the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers. Clin Cancer Res, 2000, 6(2):602-609.
23. Reuven Agami. RNAi and related mechanisms and their potential use for therapy. Current Opinion in Chemical Biology, 2002, 6: 829-834.
24. Elbashir SM, Harborth J, Lendeckel W, et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature, 2001, 411(6836):494-498
25. Ghosh SS, Gopinath P, Ramesh A. Adenoviral vectors: a promising tool for gene therapy. Appl Biochem Biotechnol, 2006, 133(1): 9-29.