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）：凋亡是针对结直肠癌的更好疗法的。 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凋亡和抑制肿瘤发生。人端粒酶逆转录酶（HTERT）启动子可以选择性地表达癌细胞中的基因。提出的研究检验以下假设：在结直肠癌细胞中通过腺病毒递送系统控制HTERT启动子下15-LOX-1表达的选择性恢复将重新凋亡并抑制结直肠癌细胞中的肿瘤发生。 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 tr​​ansfect 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水平和体外凋亡率。目的2：为了评估通过腺病毒转染系统驱动的15-LOX-1表达是否抑制了体内的结直肠瘤肿瘤发生，我们将通过肿瘤内注射到裸小鼠和15-15-效应的人类大肠癌细胞的皮下注射型细胞的皮下注射型肿瘤和皮下型皮下的肿瘤内膜。目的3：为了确定HTERT-15-15-LOX-1腺病毒的全身给药能否选择性地表达抑制肿瘤生长的肿瘤细胞中15-LOX-1，我们将对静脉内携带人类结肠癌细胞肝转移的裸小鼠进行AD-15-LOX-1，并评估15-Lox-Lox-1表达对滴虫生长的影响。目标4：为了确定15-LOX-1下调BCL-2是否诱导结直肠癌细胞中的凋亡，我们将检查（a）将AD-15-LOX-1转染到结直肠癌细胞对BCL-2表达的影响以及（如果过表达BCL-2）ripsits BCL-2诱导15 lox-1诱导的凋亡。确认肿瘤选择性靶向15-LOX-1以抑制肿瘤发生的可行性将为未来的临床前和临床发展铺平治疗策略的道路，基于分子靶向15-lox-1