Dihydrodiol dehydrogenase mediates cisplatin resistance

二氢二醇脱氢酶介导顺铂耐药

• 批准号: 7230959
• 负责人: HENRY SIMPKINS
• 金额: $ 22.29万
• 依托单位: FEINSTEIN INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7230959
• 关键词: A549; Androgens; Antibodies; Antineoplastic Agents; Apoptotic; Biological Assay; Carboplatin; Carboplatin/Cisplatin; Cell Line; Cells; Cervical; Cervix carcinoma; Cisplatin; Clinic; Clinical; Cytoskeletal Proteins; Detoxification Process; Development; Disease; Drug resistance; Enzymes; Event; Exhibits; Family; Fibroblasts; Genes; Germ cell tumor; Glucose Dehydrogenases; Glucosephosphate Dehydrogenase; Human; Immunohistochemistry; Intervention; Investigation; Length; Light; Lung; Lung Adenocarcinoma; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Messenger RNA; Microarray Analysis; Modeling; Monitor; Monoclonal Antibodies; Organ; Ovarian; Ovarian Carcinoma; Oxidoreductase; Paclitaxel; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Platinum; Population; Prostatic Neoplasms; Protein Overexpression; Proteins; Reactive Oxygen Species; Relapse; Research Personnel; Resistance; Resistance development; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Signal Transduction; Squamous cell carcinoma; Staging; Surrogate Markers; Techniques; Testing; Tissues; Transcriptional Regulation; Transfection; Treatment Protocols; Tropomyosin; Tumor Tissue; United States; Up-Regulation; Woman; Work; base; carbonyl reductase (NADPH); chemotherapy; clinically relevant; dihydrodiol dehydrogenases; enzyme activity; glucose dehydrogenase; lung Carcinoma; mRNA Expression; mortality; neoplastic cell; programs; protein expression; research study; sulfated glycoprotein 2; trans-1,2-dihydrobenzene-1,2-diol dehydrogenase; tumor

DESCRIPTION (provided by applicant): Cisplatin is one of the most effective forms of chemotherapy for advanced stage ovarian cancer although it is generally used as a carboplatin taxol combination. However some patients relapse due to the development of a resistant cell population. In order to identify clinically relevant markers for cisplatin/carboplatin responsiveness, a stable cisplatin-resistant human ovarian carcinoma cell line (2008/C13*) derived from the parental human ovarian carcinoma cell line (2008) was analyzed by microarray analysis followed by semiquantitative RT-PCR. Four genes were found to be upregulated in the cisplatin-resistant ceils: tropomyosin, apolipoprotein J (clusterin), glucose dehydrogenase and dihydro dehydrogenase. All four were upregulated by cisplatin treatment of the 2008 cells. Transfection experiments showed that forced overexpression of only DDH1 induced cisplatin/carboplatin resistance in the parental cell lines. DDH mRNA expression posttransfection was monitored by RT-PCR, and protein expression by enzyme activity assays and immunohistochemistry. Transfection of DDH1 into two other human ovarian carcinoma cell lines, 2780 and SKOV3 resulted in cisplatin carboplatin resistance, but not to other anticancer drugs. Analysis of a human lung squamous cell carcinoma cell line (A549) which has a very high levels of endogenous DDH2 mRNA exhibited a degree of cisplatin resistance greater than that of the 2008/C13* cell line. We propose to continue this work by (a) transfection of non-ovarian cell lines, including Tera-2 (germ cell tumors), A431(cervical carcinoma cells), HTB56 (lung adenocarcinoma ceils) and H69 (small cell lung carcinoma cell), to show that DDH1/2 expression produces cisplatin carboplatin resistance in a variety of cell lines derived from cancers from different organs. (b) We will also attempt to determine the transcriptional control mechanisms responsible for DDH1 upregulation as well as the (c) mechanisms by which DDH1/2 produces resistance to cisplatin (e.g. possible involvement of reactive oxygen species and apoptotic pathways involved). (d) Finally, we will study human ovarian and lung carcinoma tissues prior to and post-chemotherapy (with platinum containing drugs) utilizing monoclonal antibodies to DDH1/2 to determine if DDH expression can be used as a surrogate marker to identify whether, or not a tumor will respond to platinum-based chemotherapy.

描述（由申请人提供）： 顺铂是治疗晚期卵巢癌最有效的化疗形式之一，尽管它通常用作卡铂紫杉醇组合。然而，一些患者由于耐药细胞群的发展而复发。为了确定顺铂/卡铂反应性的临床相关标志物，通过微阵列分析和半定量 RT-PCR 分析了源自亲代人卵巢癌细胞系 (2008) 的稳定顺铂耐药人卵巢癌细胞系 (2008/C13*)。发现四个基因在顺铂耐药细胞中上调：原肌球蛋白、载脂蛋白 J（簇蛋白）、葡萄糖脱氢酶和二氢脱氢酶。 2008 细胞的顺铂处理后所有四种蛋白均上调。转染实验表明，仅强制过度表达 DDH1 可诱导亲本细胞系产生顺铂/卡铂耐药性。通过 RT-PCR 监测转染后的 DDH mRNA 表达，并通过酶活性测定和免疫组织化学监测蛋白质表达。将 DDH1 转染到另外两种人卵巢癌细胞系 2780 和 SKOV3 中会导致顺铂卡铂耐药，但不会对其他抗癌药物产生耐药性。对具有非常高水平的内源性 DDH2 mRNA 的人肺鳞状细胞癌细胞系 (A549) 的分析显示出比 2008/C13* 细胞系更高的顺铂耐药性程度。我们建议通过 (a) 转染非卵巢细胞系，包括 Tera-2（生殖细胞肿瘤）、A431（宫颈癌细胞）、HTB56（肺腺癌细胞）和 H69（小细胞肺癌细胞）来继续这项工作，以 表明 DDH1/2 表达在源自不同器官的癌症的多种细胞系中产生顺铂卡铂耐药性。 (b) 我们还将尝试确定导致 DDH1 上调的转录控制机制以及 (c) DDH1/2 产生顺铂耐药性的机制（例如可能涉及活性氧和涉及的细胞凋亡途径）。 (d) 最后，我们将利用 DDH1/2 单克隆抗体研究化疗（使用含铂药物）之前和之后的人类卵巢癌和肺癌组织，以确定 DDH 表达是否可以用作替代标记物，以确定肿瘤是否会对基于铂的化疗产生反应。

项目成果

The CCAAT box binding transcription factor, nuclear factor-Y (NF-Y) regulates transcription of human aldo-keto reductase 1C1 (AKR1C1) gene.

CCAAT 盒结合转录因子核因子 Y (NF-Y) 调节人醛酮还原酶 1C1 (AKR1C1) 基因的转录。

• DOI: 10.1016/j.gene.2010.03.006
• 发表时间: 2010
• 期刊: Gene
• 影响因子: 3.5
• 作者: Pallai,Rajash;Simpkins,Henry;Chen,Jianli;Parekh,HemantK
• 通讯作者: Parekh,HemantK

Resistance to platinum-based chemotherapy in lung cancer cell lines.

• DOI: 10.1007/s00280-010-1268-2
• 发表时间: 2010-11
• 期刊: CANCER CHEMOTHERAPY AND PHARMACOLOGY
• 影响因子: 3
• 作者: Chen, Jianli;Emara, Nashwa;Solomides, Charalambos;Parekh, Hemant;Simpkins, Henry
• 通讯作者: Simpkins, Henry