重组人P53腺病毒液对人卵巢癌细胞株生长抑制及凋亡的作用研究
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摘要
在卵巢癌中,P53基因的突变是最常见的基因改变之一。30%～79%卵巢癌中出现了P53基因的突变。目前,野生型P53基因替代疗法是肿瘤基因治疗的研究热点,而重组人P53腺病毒液则是最常用的治疗方式。有研究表明,导入野生型P53基因,能明显抑制肿瘤细胞的生长,促进细胞凋亡,增加肿瘤细胞对化疗的敏感性。因此,本课题利用人卵巢癌细胞株SKOV-3及CAOV-3,研究重组人P53腺病毒液对卵巢癌细胞的生长抑制及凋亡诱导的作用。
目的与背景:探讨研究重组人P53基因腺病毒液(rAd-P53)对卵巢腺癌细胞株的生长及化疗敏感性的影响。
方法:将rAd-P53携带的外源性P53基因导入卵巢癌细胞株SKOV-3及CAOV-3,并联合应用化疗药物DDP,通过Western blot法分析P53基因在肿瘤细胞中的表达,通过MTT法、流式细胞方法观察肿瘤细胞生长、凋亡及细胞周期的影响。
1.利用MTT方法比较不同时间、不同滴度rAd-P53对人卵巢癌细胞株CAOV-3、SKOV-3的体外生长抑制率,比较rAd-P53、DDP以及两者联合用药对卵巢肿瘤细胞的体外生长抑制率,比较联合用药时不同用药顺序对卵巢肿瘤细胞的体外生长抑制率的影响;
2.利用流式细胞方法比较rAd-P53、DDP以及两者联合用药作用于卵巢肿瘤细胞72h后肿瘤细胞的细胞周期及细胞凋亡率;
3.利用Western blot方法检测rAd-P53作用于卵巢肿瘤细胞72h后P53蛋白的表达。
结果:1.rAd-P53对人卵巢癌细胞株CAOV-3及SKOV-3的体外生长抑制作用均呈现明显的时间依赖性、剂量依赖性关系。联合用药与rAd-P53、DDP单药用药相比,对卵巢肿瘤细胞的体外生长抑制作用更显著(CAOV-3:联合用药73.57±0.43%,DDP 52.05±2.13%,rAd-P53 45.53±0.95%,P＜0.01;SKOV-3:联合用药74.70±0.86%,DDP 59.46±1.17%,rAd-P53 50.72±2.04%,P＜0.01)。联合用药时不同的用药顺序对肿瘤细胞体外生长抑制作用无明显差异(P＞0.6)。
2.联合用药72h后,肿瘤细胞的细胞周期阻滞于G0/G1期CAOV-3:67.47±1.03%,SKOV-3:67.26±2.37%,与对照组、DDP组比较有统计学差异(CAOV-3:对照组41.58±1.72%,DDP组36.87±6.53%;SKOV-3:对照组41.68±3.51%,DDP组48.36±9.34%,P＜0.01),S期比例明显减少,CAOV-3:32.51±1.06%,SKOV-3:32.53±2.32%,与对照组、DDP组、rAd-P53组比较均有统计学差异(CAOV-3:对照组54.65±2.79%,DDP组45.98±8.38%,rAd-P53组36.10±3.67%;SKOV-3:对照组54.94±0.65%,DDP组44.88±6.11%,rAd-P53组37.11±1.20%,P＜0.01),且细胞凋亡率CAOV-3达到19.39±1.67%,SKOV-3达到38.12±1.83%,与对照组、DDP组、rAd-P53组比较均有统计学差异(CAOV-3:对照组0.19±0.13%,DDP组6.61±0.49%,rAd-P53组2.61±0.20%;SKOV-3:对照组0.12±0.85%,DDP组8.52±0.47%,rAd-P53组5.70±0.47%,P＜0.01)。
3.Western blot检测rAd-P53作用于卵巢肿瘤细胞CAOV-3及SKOV-3 72h后,比较未用药物的肿瘤细胞,有明显的P53蛋白表达。
结论:rAd-P53能将外源性野生型P53基因导入肿瘤细胞的基因组,使肿瘤细胞表达P53蛋白,从而使肿瘤细胞细胞周期阻滞于G0/G1期,抑制肿瘤细胞体外生长,促进细胞凋亡。rAd-P53与常规化疗药物DDP联合应用能显著增加肿瘤细胞对DDP化疗的敏感性,其抗肿瘤效应较DDP、rAd-P53单药应用更加显著。
P53 gene is the most commonly mutated gene in ovarian tumors. There is P53 gene mutated among 30%～79% ovarian tumors. At present, wide-type P53 gene replacement therapy is among focus of tumor gene therapy, of which, recombinant adenovirus-P53 is the most commonly used. It has been reported that adenovirus-mediated wide-type P53 gene transfection into tumor cells can inhibit cell growth, promote cell apoptosis, and enhance chemotherapy sensitivity. This research is to evaluate the effects of domestic recombinant adenovirus-P53 (rAd-P53,Gendicine) on growth inhibition and apoptosis of human ovarian adenocarcinoma cell lines SKOV-3 and CAOV-3.
Objective: The aim of the study is to evaluate the effects of domestic recombinant adenovirus-P53 (rAd-P53, Gendicine) on growth and chemosensitivity of human ovarian adenocarcinoma cell lines.
Methods: Human ovarian adenocarcinoma cell lines CAOV-3 and SKOV-3 (mutant P53) were treated with rAd-P53, cisplatin (DDP) and rAd-P53 + DDP respectively. P53 expression was dectected by Western blot. The cell growth inhibition was assessed by MTT, and cell cycle and apoptosis were dectected by flow cytometry.
1. use MTT to compare CAOV-3 and SKOV-3 cell growth inhibition rate when they were treated with rAd-P53, cisplatin (DDP) and rAd-P53 + DDP respectively of different time and dose. Compare tumor cell growth inhibition of different sequence of combination treatment.
2. use flow cytometry to compare cell cycle and apoptosis of CAOV-3 and SKOV-3 when treated with rAd-P53 (100MOI), DDP (20uM) and rAd-P53 (100MOI) + DDP DDP (20uM) respectively after 72 hours.
3. use Western blot to detect P53 expression of CAOV-3 and SKOV-3 after treated with rAd-P53 (100MOI) 72 hours later.
Results: 1. There was a dose-dependent and time-dependent inhibition of cell proliferation by rAd-P53. After combined treatment with rAd-P53 (100MOI) and DDP (20uM) for 72 hours, the growth inhibition rate of CAOV-3 cells was 73.57±0.43%,which was significant higher than that in rAd-P53 group (45.53±0.95%, P<0.01) and DDP group (52.05±2.13%, P<0.01) . The growth inhibition rate of SKOV-3 cells was 74.70±0.86%, which was significant higher than that in rAd-P53 group ( 50.72±2.04%, P<0.01) and DDP group (59.46±1.17%, P<0.01) .The sequence of combined treatment is of no difference to the cellgrowth inhibition of SKOV-3 and CAOV-3.(P>0.6)
2. Combined administration of rAd-P53 and DDP remarkably arrested CAOV-3 and SKOV-3 in G0/G1 (CAOV-3: 67.47±1.03 % , SKOV-3: 67.26±2.37%), which was significantly higher than that in control group (CAOV-3: 41.58±1.72%, SKOV-3 : 41.68±3.51 %, P<0.01) and DDP group (CAOV-3: 36.87±6.53%, SKOV-3: 48.36±9.34%, P<0.01).
And cells in S phase significantly decreased (CAOV-3: 32.51±1.06%, SKOV-3: 32.53±2.32%), which was significantly lower than that in that in control group (CAOV-3: 54.65±2.79%, SKOV-3 : 54.94±0.65%, P<0.01), rAd-P53 group (CAOV-3: 36.10±3.67%, SKOV-3 : 37.11±1.20%, P<0.01) and DDP group (CAOV-3: 45.98±8.38%, SKOV-3 : 44.88±6.11%, P<0.01).
Meanwhile the apototic rate of CAOV-3 cells was 19.39±1.67% in rAd-P53 + DDP group, which was significantly higher than that in rAd-P53 group (2.61±0.20%, P<0.01) and DDP group (6.61±0.49%, P<0.01). The apototic rate of SKOV-3 cells was 38.12±1.83% in rAd-P53 + DDP group, which was significantly higher than that in rAd-P53 group (5.70±0.47%, P<0.01) and DDP group (8.52±0.47%, P<0.01).
3. High level P53 expression was dectected in CAOV-3 and SKOV-3 cells by Western blot after administeration of rAd-P53 (100MOI) 72 hours later.
Conclusion: rAd-P53 can tranfect wide-type P53 into the genome group of human ovarian tumor cells, arrest tumor cells at G0/G1, inhibit the growth of human ovarian tumor cells, promote tumor cells apoptosis. Its combibation with DDP may significantly enhance the chemosensitivity of human ovarian tumor cells to DDP.
目的与背景:探讨研究重组人P53基因腺病毒液(rAd-P53)对卵巢腺癌细胞株的生长及化疗敏感性的影响。
方法:将rAd-P53携带的外源性P53基因导入卵巢癌细胞株SKOV-3及CAOV-3,并联合应用化疗药物DDP,通过Western blot法分析P53基因在肿瘤细胞中的表达,通过MTT法、流式细胞方法观察肿瘤细胞生长、凋亡及细胞周期的影响。
1.利用MTT方法比较不同时间、不同滴度rAd-P53对人卵巢癌细胞株CAOV-3、SKOV-3的体外生长抑制率,比较rAd-P53、DDP以及两者联合用药对卵巢肿瘤细胞的体外生长抑制率,比较联合用药时不同用药顺序对卵巢肿瘤细胞的体外生长抑制率的影响;
2.利用流式细胞方法比较rAd-P53、DDP以及两者联合用药作用于卵巢肿瘤细胞72h后肿瘤细胞的细胞周期及细胞凋亡率;
3.利用Western blot方法检测rAd-P53作用于卵巢肿瘤细胞72h后P53蛋白的表达。
结果:1.rAd-P53对人卵巢癌细胞株CAOV-3及SKOV-3的体外生长抑制作用均呈现明显的时间依赖性、剂量依赖性关系。联合用药与rAd-P53、DDP单药用药相比,对卵巢肿瘤细胞的体外生长抑制作用更显著(CAOV-3:联合用药73.57±0.43%,DDP 52.05±2.13%,rAd-P53 45.53±0.95%,P＜0.01;SKOV-3:联合用药74.70±0.86%,DDP 59.46±1.17%,rAd-P53 50.72±2.04%,P＜0.01)。联合用药时不同的用药顺序对肿瘤细胞体外生长抑制作用无明显差异(P＞0.6)。
2.联合用药72h后,肿瘤细胞的细胞周期阻滞于G0/G1期CAOV-3:67.47±1.03%,SKOV-3:67.26±2.37%,与对照组、DDP组比较有统计学差异(CAOV-3:对照组41.58±1.72%,DDP组36.87±6.53%;SKOV-3:对照组41.68±3.51%,DDP组48.36±9.34%,P＜0.01),S期比例明显减少,CAOV-3:32.51±1.06%,SKOV-3:32.53±2.32%,与对照组、DDP组、rAd-P53组比较均有统计学差异(CAOV-3:对照组54.65±2.79%,DDP组45.98±8.38%,rAd-P53组36.10±3.67%;SKOV-3:对照组54.94±0.65%,DDP组44.88±6.11%,rAd-P53组37.11±1.20%,P＜0.01),且细胞凋亡率CAOV-3达到19.39±1.67%,SKOV-3达到38.12±1.83%,与对照组、DDP组、rAd-P53组比较均有统计学差异(CAOV-3:对照组0.19±0.13%,DDP组6.61±0.49%,rAd-P53组2.61±0.20%;SKOV-3:对照组0.12±0.85%,DDP组8.52±0.47%,rAd-P53组5.70±0.47%,P＜0.01)。
3.Western blot检测rAd-P53作用于卵巢肿瘤细胞CAOV-3及SKOV-3 72h后,比较未用药物的肿瘤细胞,有明显的P53蛋白表达。
结论:rAd-P53能将外源性野生型P53基因导入肿瘤细胞的基因组,使肿瘤细胞表达P53蛋白,从而使肿瘤细胞细胞周期阻滞于G0/G1期,抑制肿瘤细胞体外生长,促进细胞凋亡。rAd-P53与常规化疗药物DDP联合应用能显著增加肿瘤细胞对DDP化疗的敏感性,其抗肿瘤效应较DDP、rAd-P53单药应用更加显著。
P53 gene is the most commonly mutated gene in ovarian tumors. There is P53 gene mutated among 30%～79% ovarian tumors. At present, wide-type P53 gene replacement therapy is among focus of tumor gene therapy, of which, recombinant adenovirus-P53 is the most commonly used. It has been reported that adenovirus-mediated wide-type P53 gene transfection into tumor cells can inhibit cell growth, promote cell apoptosis, and enhance chemotherapy sensitivity. This research is to evaluate the effects of domestic recombinant adenovirus-P53 (rAd-P53,Gendicine) on growth inhibition and apoptosis of human ovarian adenocarcinoma cell lines SKOV-3 and CAOV-3.
Objective: The aim of the study is to evaluate the effects of domestic recombinant adenovirus-P53 (rAd-P53, Gendicine) on growth and chemosensitivity of human ovarian adenocarcinoma cell lines.
Methods: Human ovarian adenocarcinoma cell lines CAOV-3 and SKOV-3 (mutant P53) were treated with rAd-P53, cisplatin (DDP) and rAd-P53 + DDP respectively. P53 expression was dectected by Western blot. The cell growth inhibition was assessed by MTT, and cell cycle and apoptosis were dectected by flow cytometry.
1. use MTT to compare CAOV-3 and SKOV-3 cell growth inhibition rate when they were treated with rAd-P53, cisplatin (DDP) and rAd-P53 + DDP respectively of different time and dose. Compare tumor cell growth inhibition of different sequence of combination treatment.
2. use flow cytometry to compare cell cycle and apoptosis of CAOV-3 and SKOV-3 when treated with rAd-P53 (100MOI), DDP (20uM) and rAd-P53 (100MOI) + DDP DDP (20uM) respectively after 72 hours.
3. use Western blot to detect P53 expression of CAOV-3 and SKOV-3 after treated with rAd-P53 (100MOI) 72 hours later.
Results: 1. There was a dose-dependent and time-dependent inhibition of cell proliferation by rAd-P53. After combined treatment with rAd-P53 (100MOI) and DDP (20uM) for 72 hours, the growth inhibition rate of CAOV-3 cells was 73.57±0.43%,which was significant higher than that in rAd-P53 group (45.53±0.95%, P<0.01) and DDP group (52.05±2.13%, P<0.01) . The growth inhibition rate of SKOV-3 cells was 74.70±0.86%, which was significant higher than that in rAd-P53 group ( 50.72±2.04%, P<0.01) and DDP group (59.46±1.17%, P<0.01) .The sequence of combined treatment is of no difference to the cellgrowth inhibition of SKOV-3 and CAOV-3.(P>0.6)
2. Combined administration of rAd-P53 and DDP remarkably arrested CAOV-3 and SKOV-3 in G0/G1 (CAOV-3: 67.47±1.03 % , SKOV-3: 67.26±2.37%), which was significantly higher than that in control group (CAOV-3: 41.58±1.72%, SKOV-3 : 41.68±3.51 %, P<0.01) and DDP group (CAOV-3: 36.87±6.53%, SKOV-3: 48.36±9.34%, P<0.01).
And cells in S phase significantly decreased (CAOV-3: 32.51±1.06%, SKOV-3: 32.53±2.32%), which was significantly lower than that in that in control group (CAOV-3: 54.65±2.79%, SKOV-3 : 54.94±0.65%, P<0.01), rAd-P53 group (CAOV-3: 36.10±3.67%, SKOV-3 : 37.11±1.20%, P<0.01) and DDP group (CAOV-3: 45.98±8.38%, SKOV-3 : 44.88±6.11%, P<0.01).
Meanwhile the apototic rate of CAOV-3 cells was 19.39±1.67% in rAd-P53 + DDP group, which was significantly higher than that in rAd-P53 group (2.61±0.20%, P<0.01) and DDP group (6.61±0.49%, P<0.01). The apototic rate of SKOV-3 cells was 38.12±1.83% in rAd-P53 + DDP group, which was significantly higher than that in rAd-P53 group (5.70±0.47%, P<0.01) and DDP group (8.52±0.47%, P<0.01).
3. High level P53 expression was dectected in CAOV-3 and SKOV-3 cells by Western blot after administeration of rAd-P53 (100MOI) 72 hours later.
Conclusion: rAd-P53 can tranfect wide-type P53 into the genome group of human ovarian tumor cells, arrest tumor cells at G0/G1, inhibit the growth of human ovarian tumor cells, promote tumor cells apoptosis. Its combibation with DDP may significantly enhance the chemosensitivity of human ovarian tumor cells to DDP.
引文
[1]Mack N.Barnes,William E.Grizzle,Clinton J.Grnbbs,et al.Paradigms for Primary Prevention of Ovarian Carcinoma[J].Cancer J Clin.2002,52:216-225.
[2]Molly Brewer,Urs Utzinger,William Satterfield,et al.Biomarker Modulation in a Nonhuman Rhesus Primate Model for Ovarian Cancer Chemoprevention [J].Cancer Epidemiology,Biomarkers & Prevention.August 2001,10:889-893.
[3]Vasey PA,Shulman LN,Campos S,et al.Phase I trial ofintraperitoneal injection of the E1B-55-kd-gene-deleted adenovirus ONYX-015(dl1520)given on days 1 through 5 every 3 weeks in patients with recurrent /refractory epithelial ovarian cancer[J].J Clin Oncol,2002,20(6):1562-1569.
[4]Buller RE,Shahin MS,Horowitz JA,et al.Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500[J].CancerGeneTher,2002,9(7):567-572.
[5]Wen SF,Mahavni V,Quijano E,et al.Assessment of p53 gene transfer biological activities in a clinical study of adenovirus p53 gene therapy for recurrent ovarian cancer[J].Cancer Gene Ther,2003,10(3):224-238.
[6]Zeimet AG,Marth C.Why did p53 gene therapy fail in ovarian cancer[J].Lancet Oncol,2003,4:415-422.
[7]Zhan Q,An tinore Ⅱ,Wang XW,Carrier F,Smith ML,Harris CC Fomace AJ Jr.Association with Cdc2 and inhibition of Cdc2 / Cyclin B1 kinase activity by the p53-regulated protein Gadd45[J].Oncogene 1999,18:2892-2900.
[8]Lawler J,Miao WM,Duquette M,et al.Thrombospondin-1 gene expression affects survival and tumor spectrumof p53-deficient mice.Am J Pathol [J].2001,159:1949-1956.
[9]Frank DK,Frederick MJ,Liu TJ,et al.Bystander effect in the adenovirus-mediated wild-type p53 gene therapy model of human squamous cell carcinoma of the head and neck[J].Clin Cancer Res,1998,4:2521-2528.
[10]Wen SF,XI e I,McDonald M,et al.Development and validation of sensitive assays to quantitate gene expression after p53 gene therapy and paclitaxel chemotherapy using in vivo dosing in tumor xenograft models [J].Cancer Gene Ther,2000,7:1469-1480.
[11]Introgen's Advexin therapy halts tumor growth in advanced esophageal cancer patients[J].Introgen Therapeutics,Inc.News Release,2003,206-212.
[12]MINETA H,BONG A,DICTORM,et al.P53 mutation,but not P53expression,correlates with survival in head and neek squarous cell carcinmou[J].Br cancer,1998,78(8):1084-1090.
[13]Lorne JH,Perwez H,Curtis CH.p53:25 years after its discovery[J].Trends in Pharmacol Sci,2004,25(4):177-181.
[14]韩德民,黄志刚,张伟,等.重组人p53腺病毒注射液治疗喉癌的Ⅰ期临床试验及追踪观察[J].中华医学杂志,2003,83:2026-2029.
[15]张珊文,肖绍文,刘长清,等.重组人p53腺病毒注射液联合放射线治疗头颈鳞癌的Ⅱ期临床试验[J].中华医学杂志,2003,83:2020-2025.
[16]陈传本,潘建基,徐鹭英.重组人p53腺病毒注射液结合放疗治疗鼻咽癌Ⅱ期临床试验观察[J].中华医学杂志,2003,83:2029-2031.
[17]Peng Z.Current status of gendicine in China;recombinant human Ad-p53agent for treatment of cancers[J].Hum Gene Ther.2005,16(9):1016-1027.
[18]张毅,李叶,孟庆伟,等.野生型p53基因对卵巢癌细胞株生长及凋亡的影响[J].中国实 用妇科与产科杂志,2002,18(4):228
[19]Tin G,Zhang Y,Liu S,et al.Effects of mtDNA deletion associated with abnormal expression in rat cochlear with presbycusis[J].Clin J Biologicals,2002,15(4):225-228.
[20]Buller RE,Runnebaum IB,Karlan BY,et al.A phase Ⅰ/Ⅱ trial of rAd-p53(SCH58500)gene replacement in recurrent ovarian cancer [J].Cancer Gene Ther,2002,9(7):553-566.
[21]Vasey PA,Shulman LN,Campos S,et al.Phase I trial of intraperitoneal injection of the E1B-55-kd-gene-deleted adenovirus ONYX-015(dl1520)given on days 1 through 5 every 3 weeks in patients with recurrent/refractory epithelial ovarian cancer[J].J Clin Oncol,2002,20(6):1562-1569.
[22]Buller RE,Shahin MS,Horowitz JA,et al.Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500[J].CancerGeneTher,2002,9(7):567-572.
[23]Balachandran R,Welsh MJ,Day BW.Altered levels and regulation of stathmin in paclitaxel-resistant ovarian cancer cells[J].Oncogene,2003,22(55):8924-8930.
[24]Wolf JK,Mills GB,Bazzet L,et al.Adenovirus mediated-P53 growth inhibition of ovarian cancer cells is independent of endogenous p53 status [J].Gynecol Oncol,1999,75(2):261-266.
[25]张毅,李叶,孟庆伟,等.野生型p53基因对卵巢癌细胞株生长及凋亡的影响[J].中国实 用妇科与产科杂志,2002,18(4):228
[26]Tin G,Zhang Y,Liu S,et al.Effects of mtDNA deletion associated with abnormal expression in rat cochlear with presbycusis[J].Clin J Biologicals,2002,15(4):225-228.
[27]Kigawa J,Terakawa N.A denovirus2mediated transfer of a p53 gene in ovarian cancer[J].Adv Exp Med Biol.2000,465:207.
[28]Song K,Cowan K H,Sinha B K.In vivo studies of adenovirus-mediated p53gene therapy for cisplatinum-resistant human ovarian tumor xenografts[J].Oncol Res,1999,11(3):153.
[29]Grace MJ,Xie L,Musco ML,et al.The use of laser scanning cytometry to assess depth of penetration of adenovirus p53 gene therapy in human xenogrft biopsies[J].Am J Pathol,1999,155(6):1869-1878.
[30]Song K,Cowan KH,Sinha BK.in vivo studies ofadenovirus-mediated p53gene therapy for cisplatinum resistant human ovarian tumor xenografts [J].Oncol Res,1999,11(3):153-159.
[31]Tong XW,Kieback DG,Ramesh R,et al.Molecular aspect s of ovarian cancer.Is gene therapy the solution ?[J].ttematol Oncol Clin North Am,1999,13(1):109-133,ⅷ.
[32]Buller RE,Shahin MS,Horowitz JA,et al.Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500[J].CancerGeneTher,2002,9(7):567-572.
[1]Kato S,Han SY,Liu W,et al.Understanding the function-structure an function-mutation relationships of p53 tumor suppressor protein by high-resolution missense mutation analysis[J].PNAS,2003,100:8424-8429.
[2]Swisher SG,Roth JA.Clinical update of Ad2p53 gene therapy for lung cancer [J].Surg Oncol Clin N Am,2002,11:5212535.
[3]PrivesC,Hall PA.The p53 pathway[J].J Pathol 1999;187:111-126
[4]Zhan Q,An tinore-Ⅱ,Wang XW,Carrier F,Smith ML,Harris CC Fomace AJ Jr.Association with Cdc2 and inhibition of Cdc2/Cyclin B1 kinase activity by the p53-regulated protein Gadd45[J].Oncogene 1999:18:2892.2900
[5]Baudino TA,Cleveland JL.Th e Max network gone mad[J].Molecular Cellular Biof 2001;21:691-702
[6]Lawler J,Miao WM,Duquette M,et al.Thrombospondin-1 gene expression affects survival and tumor spectrumof p53-deficient mice[J].Am J Pathol,2001,159:1949 2 1956.
[7]M1NEYA H,BONG A,DICTORM,et al.P53 mutation,but not P53expression,correlates with survival in head and neek squarous cell carcinmou[J].Br cancer,1998,78(8):1084-1090.
[8]张爱菊.P53基因与人类肿瘤[J].医学综述,1998,4(8):405-407.
[9]Nakai H et al.Cancer Res,1992,52:6588.
[10]Levine AJ.p53,the cellular gatekeeper for growth and division[J].Cell 1997;88:323-331
[11]Komarov PG,Komarova EA,Kondratov RV,Christov-Tselkov K,Coon JS,ChemovMV,GudkovAV.A chemical inhibitor of p53 that protects mice from the side effects of cancer therapy[J].Science 1999;285:1733-1737
[12]Frank DK,Frederick MJ,Liu TJ,et al.Bystander effect in the adenovirus-mediated wild-type p53 gene therapy model of human squamous cell carcinoma of the head and neck[J].Clin Cancer Res,1998,4:252122528.
[13]Huh JJ,W olf JK,Fightmaster DI,et al.Transduction of adenovirus-mediated wild-type p53 after radiotherapy in human cervical cancer cells [J].Gynecol Oncol,2003,89:243-250.
[14]Wen SF,XI e I,McDonald M,et al.Development and validation of sensitive assays to quantitate gene expression after p53 gene therapy and paclitaxel chemotherapy using in vivo dosing in tumor xenograft models [J].Cancer Gene Ther,2000,7:1469-1480.
[15]Introgen's Advexin therapy halts tumor growth in advanced esophageal cancer patients[J].Introgen Therapeutics,Inc.News Release,2003206202.
[16]BOOKSTEIN R,DEMERS W,GREGORY R,et al.P53 gene therapy in vivo of hepatocellulur and liver metastatic colorectal cancer[J].Semin onced,1996,26:66-67.
[17]Lonardo F et al.Cancer,1997,79181:1541-1546.
[18]Schuler M,Herrlnann R,De Greve JL,et al.Adenovirus-mediated wild-type p53 gene transfer in patients receiving chemotherapy for advanced non-small-cell lung cancer:results of a multicenter phase Ⅱ study[J].J Clin Oncol.2001.19:1750-175
[19]Jenks S.Gene therapy death:everyone has to share in the guilt[J].J Natl Cancer Inst.2000.92:98-100.
[20]Bischoff JR et al.Science,1996,274:373.
[21]许少峰,付丽.p53研究的新进展[J].中华病理学杂志,2004,33:359-362.
[22]Skilling JS et al.Gynecologic Oncology,1996,60:72-78.
[23]Issaeva N,Friedler A,Bozko P,et al.Rescue of mutants of the tumor suppressor p53 in cancer cells by a designed peptide[J].Proc Natl Acad Sci USA,2003,100:13303-13307
[24]张阿丽,卢运萍,王世宣,等.卵巢癌自杀基因治疗的旁观者效应及其与间隙连接蛋白43表达的关系[J].中华妇产科杂志,2001,36(9):542-545.
[25]程月鹃,潘凌亚,张毅.重组IFN2 α 2a与HSV2tk/GCV系统协同作用增强对于卵巢癌细胞系SKOV3杀伤作用的体外研究[J].中国肿瘤生物治疗杂志,2001,8(1):13-16.
[26]Gilligan MG,Knox P,Weedon S,et al.Adenoviral delivery of B7-1(CD80)increases the immunogenicity of human ovarian and cervical carcinoma cells[J].Gene Ther,1998,5(7):965-974.
[27]Hart X,Wilbanks GD,Devaja D,et al.IL22 enhances standard IFN2 γ/L PS activation of macrophase cytotoxicity to human ovarian carcinoma in vitro:A potential for adoptive celluler immunotherapy[J].Gynecol Oncol,1999,75(2):198-210.
[28]程蓓,卵巢癌细胞因子基因治疗[J],浙江医学,2000,22(11):699
[29]Hernando JJ,Park TW,Kubler K,et al.Vaccination with autologous tumour antigen-pulsed dendritic cells in advanced gynaecological malignancies:clinical and immunological evaluation of a phase I trial[J].Cancer Immunol Immunother,2002,51(1):45-52.
[30]Buschenfelde CM,Hermann C,Schmidt B,et al.Anti human epidermal growth factor receptor 2(HER2) mono-clonal antibody trastuzumab enhances cytolytic activity of class I-restricted HER2-specific T lymphocytes against HER2-overexp ressing tumor cells[J].Cancer Research,2002,62(8):2244-7.
[31]JANICEK M F,SEVIN B U,NGU YEN H N,et al.Combination antigene therapy targeting c - myc and P53 in ovarian cancer cell lines[J]. Gynecol Oncol,1995,59:87
[32]Nakashima T,Nozawa A,Ito T,et al.Development of a new medium useful for the recovery of dermatophytes from clinical specimens by minimizing the carryover effect of antifungal agents[J].Head Neck,2002,22(5):483-488.
[33]严瑞兰,钱新华,辛晓燕,等.抗VEGF发央状核酶基因抑制VEGF表达及卵巢癌细胞增殖的实验研究[J].癌症,2002,21(1):39-44.
[34]杨红,陈志南,郑维国,等.CD147反义RNA表达质粒的构建及克隆鉴定[J].肿瘤,2002,22(3):194-196.
[35]Sood AK,Buller RE.Drug resistance in ovarian cancer from the laboratory to the clinic[J].Obstet Gynecol,1998,92:312-319.
[36]Frank TS et al.Mol Pathol,1994,7(1):2-8
[37]Teneriello MG et al.Cancer Res.1993,53(13),3103-3108
[38]Kupryjanczyk J et al.Proc Natl Acad Sci USA,1993;90:4961-4965
[39]Henriksen R et al.Gynecol Oncol,1994,53:301-306
[40]Levesque MA et al.Cancer,1995,75:1327-1338
[41]张毅,李叶,孟庆伟,等.野生型p53基因对卵巢癌细胞株生长及凋亡的影响[J].中国实用妇科与产科杂志,2002,18(4):228
[42]Balachandran R,Welsh MJ,Day BW.Altered levels and regulation of stathmin in paclitaxel-resistant ovarian cancer cells[J].Oncogene,2003,22(55):8924-8930
[43]Wolf JK,Mills GB,Bazzet L,et al.Adenovirus mediated-P53 growth inhibition of ovarian cancer cells is independent of endogenous p53 status [J].Gynecol Oncol,1999,75(2):261-266.
[44]Tin G,Zhang Y,Liu S,et al.Effects of mtDNA deletion associated with abnormal expression in rat cochlear with presbycusis[J].Clin J Biologicals,2002,15(4):225-228.
[45]Kigawa J,Terakawa N.A denovirus2mediated transfer of a p53 gene in ovarian cancer[J].Adv Exp M ed Biol.2000,465:207
[46]Song K,Cowan K H,Sinha B K.In vivo studies of adenovirus-mediated p53gene therapy for cisplatinum-resistant human ovarian tumor xenografts[J].Oncol Res.1999,11(3):153
[47]Wu Q,Chen C,Yu L,et al.Human neuronal calcium sensor-1 shows the highest expression level in cerebral cortex[J].Cancer Res Clin Oncol,2002, 126(3): 139-144.
[48] Kim J , Hwang ES , Kim J S , et al. Intraperitoneal gene therapy with adenoviral-mediated 53 tumor suppressor gene for ovarian cancer model in nude mouse [J] . Cancer Gene Ther, 1999 , 6 (2): 172-178.
[49] Buller RE, Runnebaum IB, Karlan BY, etal. A phase I / II trial of rAd-p53(SCH 8500)gene replacement in recurrent ovarian cancer[J]. Cancer Gene Ther, 2002, 9(7): 553-566.
[50] Tong XW, Kieback DG, Ramesh R, et al. Molecular aspect s of ovarian cancer. Is gene therapy the solution ? [ J ] . Hematol Oncol Clin North Am , 1999,13(1): 1092133, viii.
[51] Grace MJ, Xie L, Musco ML, et al. The use of laser scanning cytometry to assess depth of penetration of adenovirus p53 gene therapy in human xenogrft biopsies [J]. Am J Pathol, 1999, 155(6): 1869-1878.
[52] Song K, Cowan KH, Sinha BK. in vivo studies of adenovirus-mediated p53 gene therapy for cisplatinum resistant human ovarian tumor xenografts [J]. Oncol Res, 1999, 11(3): 153-159.
[53 ] Wen SF, Xie L, McDonald M, et al. Development and validation of sensitive assays to quantitate gene expression after p53 gene therapy and paclitaxel chemotherapy using in vivo dosing in tumor xenograft models [J]. Cancer Gene Ther, 2000, 7(11): 1469-1480.
[54] Shimada M, Kigawa J, Kanamori Y, et al. Mechanism of the combination effect of wild-type TP53 gene transfection and cisplatin treatment for ovarian cancer xenografts [J]. Eur J Cancer, 2000, 36(14): 1869-1875.
[55] Buller RE, Runnebaum IB, Karlan BY, et al. A phase l / II trial of rAd-p53(SCH58500)gene replacement in recurrent ovarian cancer [J]. Cancer Gene Ther, 2002, 9(7): 553-566.
[56] Vasey PA, Shulman LN, Campos S, et al. Phase I trial of intraperitoneal injection of the E1B-55-kd-gene-deleted adenovirus ONYX-015(dl1520) given on days 1 through 5 every 3 weeks in patients with recurrent / refractory epithelial ovarian cancer [J]. J Clin Oncol, 2002, 20(6): 1562-1569.
[57] Buller RE, Shahin MS, Horowitz JA, et al. Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500 [J]. CancerGeneTher, 2002, 9(7): 567-572.
[58]Wen SF,Mahavni V,Quijano E,et al.Assessment of p53 gene transfer biological activities in a clinical study of adenovirus p53 gene therapy for recurrent ovarian cancer[J].Cancer Gene Ther,2003,10(3):224-238.
[59]Zeimet AG,Marth C.Why did p53 gene therapy fail in ovarian cancer?[J].Lancet Oncol.2003,4:415-422.
[60]Tango Y,Ta M,Shirakiya Y,et al.Late resistance to adenoviral p53-mediated apoptosis caused by decreased xpression of Coxsackie-adenovirus reeeptors in human lung cancer cells[J].Cancer Sci,2004,95:459-463
[61]Zeimet A G,R iha K,Berger J,et al.New insights into p53 regulation and gene therapy for cancer[J].Biochem Pharmacol.2000,60(8):1153
[61]Bauerschm itz G J,L am J T,Kanerva A,et al.Treatment of ovarian cancer with a tropism modified oncolytic adenovirus[J].Cancer Res,2002,62:1266
[63]YoungME,Patil S,Ying J,et al.Calcitonin gene-related peptide is not essential for the development of pressure overload-induced hypertrophy in vivo[J].Bull Med Res,2001,30(3):38-40.
[64]刘春生,宋莉,孔北华,等.以非复制型HIV为载体进行目的基因转移的实验研究[J].中国医学科学院学报,2001,23(6):585-589.
[65]Hernando JJ,Park TW,Kubler K,et al.Vaccination with autologous tumour antigen-pulsed dendritic cells in advanced gynaecological malignancies:clinical and immunological evaluation of a phase I trial[J].Cancer Immunol Immunother,2002,51(1):45-52.
[2]Molly Brewer,Urs Utzinger,William Satterfield,et al.Biomarker Modulation in a Nonhuman Rhesus Primate Model for Ovarian Cancer Chemoprevention [J].Cancer Epidemiology,Biomarkers & Prevention.August 2001,10:889-893.
[3]Vasey PA,Shulman LN,Campos S,et al.Phase I trial ofintraperitoneal injection of the E1B-55-kd-gene-deleted adenovirus ONYX-015(dl1520)given on days 1 through 5 every 3 weeks in patients with recurrent /refractory epithelial ovarian cancer[J].J Clin Oncol,2002,20(6):1562-1569.
[4]Buller RE,Shahin MS,Horowitz JA,et al.Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500[J].CancerGeneTher,2002,9(7):567-572.
[5]Wen SF,Mahavni V,Quijano E,et al.Assessment of p53 gene transfer biological activities in a clinical study of adenovirus p53 gene therapy for recurrent ovarian cancer[J].Cancer Gene Ther,2003,10(3):224-238.
[6]Zeimet AG,Marth C.Why did p53 gene therapy fail in ovarian cancer[J].Lancet Oncol,2003,4:415-422.
[7]Zhan Q,An tinore Ⅱ,Wang XW,Carrier F,Smith ML,Harris CC Fomace AJ Jr.Association with Cdc2 and inhibition of Cdc2 / Cyclin B1 kinase activity by the p53-regulated protein Gadd45[J].Oncogene 1999,18:2892-2900.
[8]Lawler J,Miao WM,Duquette M,et al.Thrombospondin-1 gene expression affects survival and tumor spectrumof p53-deficient mice.Am J Pathol [J].2001,159:1949-1956.
[9]Frank DK,Frederick MJ,Liu TJ,et al.Bystander effect in the adenovirus-mediated wild-type p53 gene therapy model of human squamous cell carcinoma of the head and neck[J].Clin Cancer Res,1998,4:2521-2528.
[10]Wen SF,XI e I,McDonald M,et al.Development and validation of sensitive assays to quantitate gene expression after p53 gene therapy and paclitaxel chemotherapy using in vivo dosing in tumor xenograft models [J].Cancer Gene Ther,2000,7:1469-1480.
[11]Introgen's Advexin therapy halts tumor growth in advanced esophageal cancer patients[J].Introgen Therapeutics,Inc.News Release,2003,206-212.
[12]MINETA H,BONG A,DICTORM,et al.P53 mutation,but not P53expression,correlates with survival in head and neek squarous cell carcinmou[J].Br cancer,1998,78(8):1084-1090.
[13]Lorne JH,Perwez H,Curtis CH.p53:25 years after its discovery[J].Trends in Pharmacol Sci,2004,25(4):177-181.
[14]韩德民,黄志刚,张伟,等.重组人p53腺病毒注射液治疗喉癌的Ⅰ期临床试验及追踪观察[J].中华医学杂志,2003,83:2026-2029.
[15]张珊文,肖绍文,刘长清,等.重组人p53腺病毒注射液联合放射线治疗头颈鳞癌的Ⅱ期临床试验[J].中华医学杂志,2003,83:2020-2025.
[16]陈传本,潘建基,徐鹭英.重组人p53腺病毒注射液结合放疗治疗鼻咽癌Ⅱ期临床试验观察[J].中华医学杂志,2003,83:2029-2031.
[17]Peng Z.Current status of gendicine in China;recombinant human Ad-p53agent for treatment of cancers[J].Hum Gene Ther.2005,16(9):1016-1027.
[18]张毅,李叶,孟庆伟,等.野生型p53基因对卵巢癌细胞株生长及凋亡的影响[J].中国实 用妇科与产科杂志,2002,18(4):228
[19]Tin G,Zhang Y,Liu S,et al.Effects of mtDNA deletion associated with abnormal expression in rat cochlear with presbycusis[J].Clin J Biologicals,2002,15(4):225-228.
[20]Buller RE,Runnebaum IB,Karlan BY,et al.A phase Ⅰ/Ⅱ trial of rAd-p53(SCH58500)gene replacement in recurrent ovarian cancer [J].Cancer Gene Ther,2002,9(7):553-566.
[21]Vasey PA,Shulman LN,Campos S,et al.Phase I trial of intraperitoneal injection of the E1B-55-kd-gene-deleted adenovirus ONYX-015(dl1520)given on days 1 through 5 every 3 weeks in patients with recurrent/refractory epithelial ovarian cancer[J].J Clin Oncol,2002,20(6):1562-1569.
[22]Buller RE,Shahin MS,Horowitz JA,et al.Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500[J].CancerGeneTher,2002,9(7):567-572.
[23]Balachandran R,Welsh MJ,Day BW.Altered levels and regulation of stathmin in paclitaxel-resistant ovarian cancer cells[J].Oncogene,2003,22(55):8924-8930.
[24]Wolf JK,Mills GB,Bazzet L,et al.Adenovirus mediated-P53 growth inhibition of ovarian cancer cells is independent of endogenous p53 status [J].Gynecol Oncol,1999,75(2):261-266.
[25]张毅,李叶,孟庆伟,等.野生型p53基因对卵巢癌细胞株生长及凋亡的影响[J].中国实 用妇科与产科杂志,2002,18(4):228
[26]Tin G,Zhang Y,Liu S,et al.Effects of mtDNA deletion associated with abnormal expression in rat cochlear with presbycusis[J].Clin J Biologicals,2002,15(4):225-228.
[27]Kigawa J,Terakawa N.A denovirus2mediated transfer of a p53 gene in ovarian cancer[J].Adv Exp Med Biol.2000,465:207.
[28]Song K,Cowan K H,Sinha B K.In vivo studies of adenovirus-mediated p53gene therapy for cisplatinum-resistant human ovarian tumor xenografts[J].Oncol Res,1999,11(3):153.
[29]Grace MJ,Xie L,Musco ML,et al.The use of laser scanning cytometry to assess depth of penetration of adenovirus p53 gene therapy in human xenogrft biopsies[J].Am J Pathol,1999,155(6):1869-1878.
[30]Song K,Cowan KH,Sinha BK.in vivo studies ofadenovirus-mediated p53gene therapy for cisplatinum resistant human ovarian tumor xenografts [J].Oncol Res,1999,11(3):153-159.
[31]Tong XW,Kieback DG,Ramesh R,et al.Molecular aspect s of ovarian cancer.Is gene therapy the solution ?[J].ttematol Oncol Clin North Am,1999,13(1):109-133,ⅷ.
[32]Buller RE,Shahin MS,Horowitz JA,et al.Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500[J].CancerGeneTher,2002,9(7):567-572.
[1]Kato S,Han SY,Liu W,et al.Understanding the function-structure an function-mutation relationships of p53 tumor suppressor protein by high-resolution missense mutation analysis[J].PNAS,2003,100:8424-8429.
[2]Swisher SG,Roth JA.Clinical update of Ad2p53 gene therapy for lung cancer [J].Surg Oncol Clin N Am,2002,11:5212535.
[3]PrivesC,Hall PA.The p53 pathway[J].J Pathol 1999;187:111-126
[4]Zhan Q,An tinore-Ⅱ,Wang XW,Carrier F,Smith ML,Harris CC Fomace AJ Jr.Association with Cdc2 and inhibition of Cdc2/Cyclin B1 kinase activity by the p53-regulated protein Gadd45[J].Oncogene 1999:18:2892.2900
[5]Baudino TA,Cleveland JL.Th e Max network gone mad[J].Molecular Cellular Biof 2001;21:691-702
[6]Lawler J,Miao WM,Duquette M,et al.Thrombospondin-1 gene expression affects survival and tumor spectrumof p53-deficient mice[J].Am J Pathol,2001,159:1949 2 1956.
[7]M1NEYA H,BONG A,DICTORM,et al.P53 mutation,but not P53expression,correlates with survival in head and neek squarous cell carcinmou[J].Br cancer,1998,78(8):1084-1090.
[8]张爱菊.P53基因与人类肿瘤[J].医学综述,1998,4(8):405-407.
[9]Nakai H et al.Cancer Res,1992,52:6588.
[10]Levine AJ.p53,the cellular gatekeeper for growth and division[J].Cell 1997;88:323-331
[11]Komarov PG,Komarova EA,Kondratov RV,Christov-Tselkov K,Coon JS,ChemovMV,GudkovAV.A chemical inhibitor of p53 that protects mice from the side effects of cancer therapy[J].Science 1999;285:1733-1737
[12]Frank DK,Frederick MJ,Liu TJ,et al.Bystander effect in the adenovirus-mediated wild-type p53 gene therapy model of human squamous cell carcinoma of the head and neck[J].Clin Cancer Res,1998,4:252122528.
[13]Huh JJ,W olf JK,Fightmaster DI,et al.Transduction of adenovirus-mediated wild-type p53 after radiotherapy in human cervical cancer cells [J].Gynecol Oncol,2003,89:243-250.
[14]Wen SF,XI e I,McDonald M,et al.Development and validation of sensitive assays to quantitate gene expression after p53 gene therapy and paclitaxel chemotherapy using in vivo dosing in tumor xenograft models [J].Cancer Gene Ther,2000,7:1469-1480.
[15]Introgen's Advexin therapy halts tumor growth in advanced esophageal cancer patients[J].Introgen Therapeutics,Inc.News Release,2003206202.
[16]BOOKSTEIN R,DEMERS W,GREGORY R,et al.P53 gene therapy in vivo of hepatocellulur and liver metastatic colorectal cancer[J].Semin onced,1996,26:66-67.
[17]Lonardo F et al.Cancer,1997,79181:1541-1546.
[18]Schuler M,Herrlnann R,De Greve JL,et al.Adenovirus-mediated wild-type p53 gene transfer in patients receiving chemotherapy for advanced non-small-cell lung cancer:results of a multicenter phase Ⅱ study[J].J Clin Oncol.2001.19:1750-175
[19]Jenks S.Gene therapy death:everyone has to share in the guilt[J].J Natl Cancer Inst.2000.92:98-100.
[20]Bischoff JR et al.Science,1996,274:373.
[21]许少峰,付丽.p53研究的新进展[J].中华病理学杂志,2004,33:359-362.
[22]Skilling JS et al.Gynecologic Oncology,1996,60:72-78.
[23]Issaeva N,Friedler A,Bozko P,et al.Rescue of mutants of the tumor suppressor p53 in cancer cells by a designed peptide[J].Proc Natl Acad Sci USA,2003,100:13303-13307
[24]张阿丽,卢运萍,王世宣,等.卵巢癌自杀基因治疗的旁观者效应及其与间隙连接蛋白43表达的关系[J].中华妇产科杂志,2001,36(9):542-545.
[25]程月鹃,潘凌亚,张毅.重组IFN2 α 2a与HSV2tk/GCV系统协同作用增强对于卵巢癌细胞系SKOV3杀伤作用的体外研究[J].中国肿瘤生物治疗杂志,2001,8(1):13-16.
[26]Gilligan MG,Knox P,Weedon S,et al.Adenoviral delivery of B7-1(CD80)increases the immunogenicity of human ovarian and cervical carcinoma cells[J].Gene Ther,1998,5(7):965-974.
[27]Hart X,Wilbanks GD,Devaja D,et al.IL22 enhances standard IFN2 γ/L PS activation of macrophase cytotoxicity to human ovarian carcinoma in vitro:A potential for adoptive celluler immunotherapy[J].Gynecol Oncol,1999,75(2):198-210.
[28]程蓓,卵巢癌细胞因子基因治疗[J],浙江医学,2000,22(11):699
[29]Hernando JJ,Park TW,Kubler K,et al.Vaccination with autologous tumour antigen-pulsed dendritic cells in advanced gynaecological malignancies:clinical and immunological evaluation of a phase I trial[J].Cancer Immunol Immunother,2002,51(1):45-52.
[30]Buschenfelde CM,Hermann C,Schmidt B,et al.Anti human epidermal growth factor receptor 2(HER2) mono-clonal antibody trastuzumab enhances cytolytic activity of class I-restricted HER2-specific T lymphocytes against HER2-overexp ressing tumor cells[J].Cancer Research,2002,62(8):2244-7.
[31]JANICEK M F,SEVIN B U,NGU YEN H N,et al.Combination antigene therapy targeting c - myc and P53 in ovarian cancer cell lines[J]. Gynecol Oncol,1995,59:87
[32]Nakashima T,Nozawa A,Ito T,et al.Development of a new medium useful for the recovery of dermatophytes from clinical specimens by minimizing the carryover effect of antifungal agents[J].Head Neck,2002,22(5):483-488.
[33]严瑞兰,钱新华,辛晓燕,等.抗VEGF发央状核酶基因抑制VEGF表达及卵巢癌细胞增殖的实验研究[J].癌症,2002,21(1):39-44.
[34]杨红,陈志南,郑维国,等.CD147反义RNA表达质粒的构建及克隆鉴定[J].肿瘤,2002,22(3):194-196.
[35]Sood AK,Buller RE.Drug resistance in ovarian cancer from the laboratory to the clinic[J].Obstet Gynecol,1998,92:312-319.
[36]Frank TS et al.Mol Pathol,1994,7(1):2-8
[37]Teneriello MG et al.Cancer Res.1993,53(13),3103-3108
[38]Kupryjanczyk J et al.Proc Natl Acad Sci USA,1993;90:4961-4965
[39]Henriksen R et al.Gynecol Oncol,1994,53:301-306
[40]Levesque MA et al.Cancer,1995,75:1327-1338
[41]张毅,李叶,孟庆伟,等.野生型p53基因对卵巢癌细胞株生长及凋亡的影响[J].中国实用妇科与产科杂志,2002,18(4):228
[42]Balachandran R,Welsh MJ,Day BW.Altered levels and regulation of stathmin in paclitaxel-resistant ovarian cancer cells[J].Oncogene,2003,22(55):8924-8930
[43]Wolf JK,Mills GB,Bazzet L,et al.Adenovirus mediated-P53 growth inhibition of ovarian cancer cells is independent of endogenous p53 status [J].Gynecol Oncol,1999,75(2):261-266.
[44]Tin G,Zhang Y,Liu S,et al.Effects of mtDNA deletion associated with abnormal expression in rat cochlear with presbycusis[J].Clin J Biologicals,2002,15(4):225-228.
[45]Kigawa J,Terakawa N.A denovirus2mediated transfer of a p53 gene in ovarian cancer[J].Adv Exp M ed Biol.2000,465:207
[46]Song K,Cowan K H,Sinha B K.In vivo studies of adenovirus-mediated p53gene therapy for cisplatinum-resistant human ovarian tumor xenografts[J].Oncol Res.1999,11(3):153
[47]Wu Q,Chen C,Yu L,et al.Human neuronal calcium sensor-1 shows the highest expression level in cerebral cortex[J].Cancer Res Clin Oncol,2002, 126(3): 139-144.
[48] Kim J , Hwang ES , Kim J S , et al. Intraperitoneal gene therapy with adenoviral-mediated 53 tumor suppressor gene for ovarian cancer model in nude mouse [J] . Cancer Gene Ther, 1999 , 6 (2): 172-178.
[49] Buller RE, Runnebaum IB, Karlan BY, etal. A phase I / II trial of rAd-p53(SCH 8500)gene replacement in recurrent ovarian cancer[J]. Cancer Gene Ther, 2002, 9(7): 553-566.
[50] Tong XW, Kieback DG, Ramesh R, et al. Molecular aspect s of ovarian cancer. Is gene therapy the solution ? [ J ] . Hematol Oncol Clin North Am , 1999,13(1): 1092133, viii.
[51] Grace MJ, Xie L, Musco ML, et al. The use of laser scanning cytometry to assess depth of penetration of adenovirus p53 gene therapy in human xenogrft biopsies [J]. Am J Pathol, 1999, 155(6): 1869-1878.
[52] Song K, Cowan KH, Sinha BK. in vivo studies of adenovirus-mediated p53 gene therapy for cisplatinum resistant human ovarian tumor xenografts [J]. Oncol Res, 1999, 11(3): 153-159.
[53 ] Wen SF, Xie L, McDonald M, et al. Development and validation of sensitive assays to quantitate gene expression after p53 gene therapy and paclitaxel chemotherapy using in vivo dosing in tumor xenograft models [J]. Cancer Gene Ther, 2000, 7(11): 1469-1480.
[54] Shimada M, Kigawa J, Kanamori Y, et al. Mechanism of the combination effect of wild-type TP53 gene transfection and cisplatin treatment for ovarian cancer xenografts [J]. Eur J Cancer, 2000, 36(14): 1869-1875.
[55] Buller RE, Runnebaum IB, Karlan BY, et al. A phase l / II trial of rAd-p53(SCH58500)gene replacement in recurrent ovarian cancer [J]. Cancer Gene Ther, 2002, 9(7): 553-566.
[56] Vasey PA, Shulman LN, Campos S, et al. Phase I trial of intraperitoneal injection of the E1B-55-kd-gene-deleted adenovirus ONYX-015(dl1520) given on days 1 through 5 every 3 weeks in patients with recurrent / refractory epithelial ovarian cancer [J]. J Clin Oncol, 2002, 20(6): 1562-1569.
[57] Buller RE, Shahin MS, Horowitz JA, et al. Long term follow-up of patients with recurrent ovarian cancer after Ad-p53 gene replacement with SCH 58500 [J]. CancerGeneTher, 2002, 9(7): 567-572.
[58]Wen SF,Mahavni V,Quijano E,et al.Assessment of p53 gene transfer biological activities in a clinical study of adenovirus p53 gene therapy for recurrent ovarian cancer[J].Cancer Gene Ther,2003,10(3):224-238.
[59]Zeimet AG,Marth C.Why did p53 gene therapy fail in ovarian cancer?[J].Lancet Oncol.2003,4:415-422.
[60]Tango Y,Ta M,Shirakiya Y,et al.Late resistance to adenoviral p53-mediated apoptosis caused by decreased xpression of Coxsackie-adenovirus reeeptors in human lung cancer cells[J].Cancer Sci,2004,95:459-463
[61]Zeimet A G,R iha K,Berger J,et al.New insights into p53 regulation and gene therapy for cancer[J].Biochem Pharmacol.2000,60(8):1153
[61]Bauerschm itz G J,L am J T,Kanerva A,et al.Treatment of ovarian cancer with a tropism modified oncolytic adenovirus[J].Cancer Res,2002,62:1266
[63]YoungME,Patil S,Ying J,et al.Calcitonin gene-related peptide is not essential for the development of pressure overload-induced hypertrophy in vivo[J].Bull Med Res,2001,30(3):38-40.
[64]刘春生,宋莉,孔北华,等.以非复制型HIV为载体进行目的基因转移的实验研究[J].中国医学科学院学报,2001,23(6):585-589.
[65]Hernando JJ,Park TW,Kubler K,et al.Vaccination with autologous tumour antigen-pulsed dendritic cells in advanced gynaecological malignancies:clinical and immunological evaluation of a phase I trial[J].Cancer Immunol Immunother,2002,51(1):45-52.