Molecular Targeting of 15-Lipoxygenase-1 in Colon Cancer

15-脂氧合酶-1 在结肠癌中的分子靶向

• 批准号: 7103666
• 负责人: Imad Shureiqi
• 金额: $ 23.22万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7103666
• 关键词: Adenoviridae; BCL2 gene /protein; apoptosis; athymic mouse; biotechnology; carcinogenesis; cell line; cell proliferation; colorectal neoplasms; enzyme linked immunosorbent assay; gene expression; gene therapy; genetic promoter element; linoleate; lipoxygenase; metastasis; neoplasm /cancer therapy; neoplasm /cancer transplantation; neoplastic cell; neoplastic growth; telomerase; therapy design /development; transfection /expression vector; xenotransplantation

DESCRIPTION (provided by applicant): Apoptosis is being targeted to develop better therapies for colorectal cancer. We have found that (a) linoleic acid's product through 15-1ipoxygenase-1 (15-LOX-1), 13-S-hydroxyoctadecadienoic acid (13-S-HODE), restores apoptosis in colorectal cancer cells, (b) 13-S-HODE and 15-LOX-1 are downregulated in human colorectal cancer, and (c) NSAIDs restore 15-LOX-1 expression in human colorectal cancer cells to induce apoptosis and inhibit tumorigenesis. Human telomerase reverse transcriptase (hTERT) promoter can selectively express genes in cancer cells. The proposed research tests the following hypothesis: Selective restoration of 15-LOX-1 expression under the control of hTERT promoter in colorectal cancer cells via an adenoviral delivery system will reestablish apoptosis and inhibit tumorigenesis in colorectal cancer cells. Specific Aims: Aim 1: To determine whether hTERT-driven 15-LOX-1 expression via an adenoviral vector reestablishes apoptosis through the expression of 15-LOX-1 in colorectal cancer cells, we will transfect colorectal cancer cell lines with an adenoviral vector (Ad-15-LOX-1) that expresses 15-LOX-1 under the control of the hTERT promoter and will measure 15-LOX-1 expression, 13-S-HODE levels, and apoptosis rates in vitro. Aim 2: To assess whether 15-LOX-1 expression driven by hTERT promoter through an adenoviral transfection system suppresses colorectal tumorigenesis in vivo, we will transfect Ad-15-LOX-1 by intratumoral injections into subcutaneous xenografts of human colorectal cancer cells in nude mice and assess the effects of 15-LOX-1 expression on tumorigenesis and apoptosis rates. Aim 3: To determine whether the systemic administration of hTERT-15-LOX-1 adenovirus selectively expresses 15-LOX-1 in tumor cells suppressing tumor growth, we will administer Ad-15-LOX-1 intravenously to nude mice carrying human colon cancer cell liver metastases and evaluate the 15-LOX-1 expression effects on tumor growth. Aim 4: To determine whether 15-LOX-1 downregulates Bcl-2 to induce apoptosis in colorectal cancer cells, we will examine (a) the effects of transfecting Ad-15-LOX-1 into colorectal cancer cells on Bcl-2 expression and (b) if overexpressing Bcl-2 inhibits 15-LOX-1 induced apoptosis. Confirming the feasibility of tumor selective targeting of 15-LOX-1 to inhibit tumorigenesis will pave the way for future preclinical and clinical development of therapeutic strategies based on molecularly targeting 15-LOX-1

描述（由申请人提供）：靶向细胞凋亡以开发更好的结直肠癌治疗方法。我们已经发现（a）亚油酸通过15-脂氧合酶-1（15-LOX-1）的产物13-S-羟基十八碳二烯酸（13-S-HODE）恢复结肠直肠癌细胞中的细胞凋亡，（B）13-S-HODE和15-LOX-1在人结肠直肠癌中下调，和（c）NSAID恢复人结肠直肠癌细胞中的15-LOX-1表达以诱导细胞凋亡和抑制肿瘤发生。人端粒酶逆转录酶（Human telomerase reverse transcriptase，hTERT）启动子可在肿瘤细胞中选择性表达基因。该研究验证了以下假设：通过腺病毒递送系统在结肠直肠癌细胞中选择性恢复hTERT启动子控制下的15-LOX-1表达将重建结肠直肠癌细胞的凋亡并抑制肿瘤发生。具体目标：目标1：为了确定通过腺病毒载体hTERT驱动的15-LOX-1表达是否通过15-LOX-1在结肠直肠癌细胞中的表达重建凋亡，我们将用腺病毒载体转染结肠直肠癌细胞系，（Ad-15-LOX-1），其在hTERT启动子的控制下表达15-LOX-1，并将测量15-LOX-1表达，13-S-HODE水平，和细胞凋亡率。目标二：为了评估由hTERT启动子通过腺病毒转染系统驱动的15-LOX-1表达是否抑制体内结直肠肿瘤发生，我们将通过瘤内注射到裸鼠皮下移植的人结直肠癌细胞中来检测Ad-15-LOX-1，并评估15-LOX-1表达对肿瘤发生和凋亡率的影响。目标三：为了确定hTERT-15-LOX-1腺病毒的全身给药是否在抑制肿瘤生长的肿瘤细胞中选择性地表达15-LOX-1，我们将向携带人结肠癌细胞肝转移的裸鼠静脉内给药Ad-15-LOX-1，并评估15-LOX-1表达对肿瘤生长的影响。目标4：为了确定15-LOX-1是否下调Bcl-2以诱导结肠直肠癌细胞中的凋亡，我们将检测（a）将Ad-15-LOX-1转染到结肠直肠癌细胞中对Bcl-2表达的影响和（B）过表达Bcl-2是否抑制15-LOX-1诱导的凋亡。阐明肿瘤选择性靶向15-LOX-1抑制肿瘤发生的可行性将为未来基于分子靶向15-LOX-1的治疗策略的临床前和临床开发铺平道路