Molecular Analysis of Resistance to p53 Gene Therapy in Human Lung Cancer
Molecular Analysis of Resistance to p53 Gene Therapy in Human Lung Cancer
批准号:
13671390
负责人:
FUJIWARA Toshiyoshi
金额:
$2.62万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (C)
财政年份:
2001
资助国家:
日本
项目状态:
已结题
起止时间:
2001 至 2002
中文摘要
为了分析在体内表达p53肿瘤抑制因子(Ad-p53)的腺病毒载体的抗肿瘤作用机制,我们定量评估了p53靶向基因在经Ad-p53处理的裸鼠肿瘤中的表达,并可视化了p53的转录活性。裸鼠建立的人肺癌(H1299)异种移植物瘤内给药Ad-p53治疗。实时逆转录聚合酶链反应(RT-PCR)检测外源性p53及p53靶基因p21、MDM2、Noxa、p53AIP1的表达水平,并于治疗后第1、2、3、7、14天组织化学观察细胞凋亡诱导情况。除p53AIP1外,外源p53和p53靶基因mrna表达在Ad-p53治疗后1天达到最大值,随后迅速下降;治疗后2 ~ 3天细胞凋亡明显。我们开发了一种非侵入性和简单的方法来监测裸鼠体内Ad-p53的转录活性。我们建立了在p53-responsive p21 promoter(即p53R-GFP报告基因系统)控制下表达绿色荧光蛋白(GFP)报告基因的H1299细胞。将这些细胞移植到裸鼠体内,瘤内注射Ad-p53,通过3-CCD_camera实时观察外源p53的转录活性和瘤内GFP的表达。GFP在治疗后第3天表达量最大,在治疗后第7天显著降低。我们证明了Ad-p53治疗可以快速诱导p53靶向基因和肿瘤细胞凋亡。我们还成功地可视化了p53在体内的转录活性。利用实时定量RT-PCR技术定量分析p53靶向基因表达,利用p53R-GFP报告系统可视化新鲜异种移植物中p53转录活性,可能有助于评估Ad-pS3抗肿瘤作用的机制和新的治疗方法。少
英文摘要
To analyze the mechanism of the antitumor effect of an adenoviral vector expressing the p53 tumor suppressor (Ad-p53) in vivo, we quantitatively assessed p53-targeted gene expression and visualized transcriptional activity of p53 in tumors in nude mice treated with Ad-p53. Human lung cancer (H1299) xenografts established in nude mice were treated by intratumoral administration of Ad-p53. The levels of expression of exogenous p53 and p53-targeted genes p21, MDM2, Noxa, and p53AIP1 were quantified by real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and induction of apoptosis was observed histochemically on days 1, 2, 3, 7 and 14 after treatment. Expression of mRNAs of exogenous p53 and p53-targeted genes (except p53AIP1) was at its maximum 1 day after Ad-p53 treatment, then decreased rapidly ; apoptosis was evident in situ 2-3 days after treatment. We developed a noninvasive and simple method for monitoring the transcriptional activity of exogenous p53 following intratu … More moral administration of Ad-p53 in nude mice. We established H1299 cells that express the green fluorescent protein (GFP) reporter gene under the control of p53-responsive p21 promotes (i.e., the p53R-GFP reporter system). Xenografts of these cells in nude mice were treated by intratumoral administration of Ad-p53, and the transcriptional activity of exogenous p53 could be visualized as intratumoral GFP expression in real time by 3-CCD_camera. Expression of GFP was maximal 3 days after treatment, and it decreased remarkably by 7 days after treatment. We demonstrated that Ad-p53 treatment rapidly induced p53-targeted genes and apoptosis in tumors. We also succeeded in visualizing p53 transcriptional activity in vivo. Quantitative analysis of p53-targeted gene expression by real-time quantitative RT-PCR and visualization of p53 transcriptional activity in fresh xenografts by using the p53R-GFP reporter system may be useful in assessing the mechanisms of the antitumor effects of Ad-pS3 and novel therapeutic approaches. Less
期刊论文(5)
专著(0)
科研奖励(0)
会议论文
登录
查看更多内容
Shao, J., Fujiwara, T., et al.: "p53 inhibits adriamycin-induced down-regulation of cyclin D1 expression in human cancer cells"Biochem.Bioph.Res.Co.. 290. 1101-1107 (2002)
Shao,J.,Fujiwara,T.,等人:“p53 抑制阿霉素诱导的人癌细胞中细胞周期蛋白 D1 表达的下调”Biochem.Bioph.Res.Co.. 290. 1101-1107 (2002)
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Teraishi, F., Fujiwara, T., et al.: "Ectopic p21^<sdi1> gene transfer induces retinoic acid receptor beta expression and sensitizes human cancer cells to retinoid treatment"Int.J.Cancer. 103. 833-839 (2003)
Teraishi, F.、Fujiwara, T. 等人:“异位 p21^<sdi1> 基因转移诱导视黄酸受体 β 表达并使人类癌细胞对类视黄醇治疗敏感”Int.J.Cancer。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Ohtani S, Fujiwara T, et al.: "Quantitative analysis of p53-targeted gene expression and visualization of p53 trans-criptional activity following intratumoral administration of adenoviral p53 in vivo."Mol.Cancer Ther.. 3. 93-100 (2004)
Ohtani S、Fujiwara T 等人:“体内腺病毒 p53 瘤内给药后 p53 靶向基因表达的定量分析和 p53 转录活性的可视化。”Mol.Cancer Ther.. 3. 93-100 (2004
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Ohtani, S., Kagawa, S., Tnago, Y., Umeoka, T., Tokunaga, N., Tsunemitsu, Y., Roth, J.A., Taya, Y.S., Tanaka, N., Fujiwara, T.: "Quantitative analysis of p53-targeted gene expression and visualization of p53 transcriptional activity following intratumoral
Ohtani, S.、Kakawa, S.、Tnago, Y.、Umeoka, T.、Tokunaga, N.、Tsunemitsu, Y.、Roth, J.A.、Taya, Y.S.、Tanaka, N.、Fujiwara, T.:“定量
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
藤原俊義, 田中紀章: "遺伝子治療「TECHNICAL TERM緩和医療」"先端医学社. 172-173 (2002)
Toshiyoshi Fujiwara、Kisho Tanaka:“基因疗法‘技术术语姑息医学’”Senshin Igakusha 172-173 (2002)。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
Development of the Dual fluorescent cytology with tumor-specific viruses for gastrointestinal cancer precision medicine
-
批准号:16H05416
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$11.32万
-
财政年份:2016
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
Fluorescent virus-guided capturing of epithelial-mesenchymal transition (EMT)-induced circulating tumor cells (CTCs) for genetic analysis
-
批准号:25670580
-
项目类别:Grant-in-Aid for Challenging Exploratory Research
-
资助金额:$2.41万
-
财政年份:2013
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
"Cell-in-cell" activity-based tumor-specific delivery of biologics
-
批准号:25293283
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$11.73万
-
财政年份:2013
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
Molecular therapy for gastric cancer using HER2-extracellular domain-expressing adenovirus
-
批准号:22390256
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$12.4万
-
财政年份:2010
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
Telomerase-specific virotherapy for malignant mesothelioma
-
批准号:19390365
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$9.98万
-
财政年份:2007
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
Telomerase-specific fluorescent navigation system for lung cancer surgery
-
批准号:17390381
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$9.86万
-
财政年份:2005
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
Molecular Fluorescent Imaging of Metastatic Tumors with Conditionally Replication-Selective Adenovirus and GFP Gene
-
批准号:15591475
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$2.24万
-
财政年份:2003
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
Development of Gene Therapy for Lung Cancer using a Novel Antiangiogenic Factor BAI1
-
批准号:11671325
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$2.24万
-
财政年份:1999
-
负责人:FUJIWARA Toshiyoshi
-
依托单位:
海外基金