Molecular Cytogenetics of Solid Tumors

实体瘤的分子细胞遗传学

基本信息

批准号：
7732973
负责人：
NICOLAE POPESCU
金额：
$ 98.51万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7732973
关键词：
Actins Adenovirus Vector Adenoviruses Age Aging Aging-Related Process Androgens Apoptosis Apoptosis Promoter Breast Cancer Etiology Cancer Prognosis Cell Aging Cell Cycle Arrest Cell Death Cell Migration Inhibition function Cell Migration Inhibition measurement Cell Nucleus Cell Proliferation Cells Cellular Morphology Chemicals Clinical Trials Complex Cytoplasm Cytoskeleton Cytoskeleton Alteration DLC1 gene DNA Damage DNA Double Strand Break DNA Repair Development Diagnosis Disease Distant Double Strand Break Repair Down-Regulation Epigenetic Process Event Exhibits Fibroblasts Gelatinase B Gene Expression Gene Transfer Genes Genome Stability Genomic Instability Genomics Goals Growth Histone Deacetylase Inhibitor Histones Human Immigration In Vitro Incidence Individual Induction of Apoptosis Lead Link Liver Liver neoplasms Lung Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of liver Malignant neoplasm of lung Malignant neoplasm of prostate Mediating Metastasis Suppressor Genes Metastatic Prostate Cancer Molecular Molecular Biology Molecular Cytogenetics Mutation Nature Neoplasm Metastasis Normal Cell Nuclear Translocation Nude Mice Organ Patients Play Premature aging syndrome Prevention Primary Neoplasm Primary carcinoma of the liver cells Process Prognostic Marker Prostatic Neoplasms Proteins Range Rate Recurrence Research Resistance Role Screening for Prostate Cancer Signal Pathway Site Solid Neoplasm Stress Fibers Testing Therapeutic Intervention Tumor Suppressor Genes Tumor Suppressor Proteins Tumorigenicity Werner Syndrome age related base cancer cell cancer therapy dietary constituent drug development gene function gene therapy human DLC1 protein human H2AX protein in vivo inhibitor/antagonist lung Carcinoma mortality neoplastic cell novel osteopontin outcome forecast pathological aging prevent programs protein expression response restoration senescence tumor tumor progression

项目摘要

In the past year significant progress has been made in understanding the role of DNA damage in cellular aging and senescence and in the understanding of the Deleted in Liver Cancer 1, (DLC1), genes tumor suppressive function and its potential use for therapeutic interventions. Accumulation of DNA damage may play an essential role not only in the process of aging but also in induction of cellular senescence. The ability of cells to sense and repair DNA damage declines with age. However, the underlying molecular mechanism for this age-dependent decline is still elusive. To understand quantitative and qualitative changes in the DNA damage response during human aging, the nature and dynamics of phosphorylated histone H2AX (gamma-H2AX) foci, which occur specifically at sites of DNA double-strand breaks (DSB), were examined in cells from normal individuals of different age and from patients with Werner syndrome (WS), a disorder associated with premature aging, genomic instability and increased incidence of cancer. This study showed that the incidence of gamma-H2AX foci generally increases with donors age, although the cells derived from patients with Werner syndrome (WS) exhibited considerably higher incidence of gamma-H2AX foci than those taken from normal donors of comparable age. Further increases in gamma-H2AX focal incidence occurred in culture as both normal and WS fibroblasts progressed toward senescence. The rates of recruitment of DSB repair proteins to gamma-H2AX foci correlated inversely with age for both normal and WS donors, perhaps due in part to the slower growth of gamma-H2AX foci in older donors. Since genomic stability may depend on the efficient processing of DSBs where the rapid formation of gamma-H2AX foci and the rapid accumulation of DSB repair proteins may be instrumental, our findings suggest that increasing inefficiency in these processes may contribute to genome instability associated with normal and pathological aging. The DLC1 gene, isolated and characterized by our group, has recently been independently confirmed as a potent bona fide tumor suppressor gene. Silencing or underexpression of this gene plays an important role in several common human cancers such as liver, breast, lung and prostate cancer and, most significantly, signaling pathways modulated by DLC1 provide wide range of targets for therapeutic interventions. Induction of apoptosis is a major attribute of tumor suppressor gene. In two independent studies with lung and prostate tumor cells, we identified cellular and molecular alterations indicative of the induction of programmed cell death by DLC1. DLC1 has a dramatic inhibitory effect on in vitro cell proliferation and prevented in vivo the development of tumors after inoculation of human non-small cell lung carcinoma cells in nude mice. Transduction of DLC1 in lung cancer cells invariably induced changes in cell morphology corroborative of the tumor suppressor function of the gene. DLC1-mediated actin cytoskeleton-based morphological alterations, and DLC1 protein nuclear translocation occur prior to inhibition of cell migration and before to the induction of apoptosis. These observations led to the conclusion that DLC1 functions in the cytoplasm as an inhibitor of tumor cell proliferation and migration, but in the nucleus as an inducer of apoptosis. Based on our evidence, DLC1 is now considered a metastasis suppressor gene. The process of tumor cell dissemination from the locally growing primary tumors to metastasis in anatomically distant sites is the leading cause of cancer mortality, representing over 90% of cancer casualties. Metastatic disease is a complex process involving various cellular and genetic alterations. We showed that DLC1 inhibits the spread of liver cancer cells to distant organs. The inhibition of liver cancer cell dissemination to distant organs was associated with reduction of RhoA activity, cytoskeleton alterations, down regulation of osteopontin (OPN) and matrix metalloproteinase-9 (MMP9) expression, which are highly up regulated in most primary liver tumors with associated metastases. Over-expression of OPN and MMP-9 and silencing of DLC1 may provide a prognostic marker for unfavorable prognosis of liver cancer. DLC1 gene therapy using adenoviral vector, as the most useful vehicle for gene transfer, is a realistic prospect for prevention of HCC dissemination. Also, searching for dietary constituents and pharmacological agents that up-regulate DLC-1 gene expression could lead to the development of drugs that might be useful for prevention of metastasis, the most harmful event during liver cancer progression. In prostate cancer, we found that recurrent downregulation and inactivation of DLC1 by epigenetic mechanisms may serve as an marker for early detection of prostate cancer and subsequently we showed that adenovirus-mediated DLC1 transfer in androgen-independent aggressive and metastatic prostate tumor cells inhibited cell proliferation in vitro, their tumorigenicity in nude mice, and induced apoptosis. It also induced cell cycle arrest, inhibited the activation of RhoA and the formation of actin stress fibers. In prostate tumor cells that are resistant to histone deacetylase inhibitors-induced cell death, restoration of DLC1 expression induced apoptosis only after chemical inhibition of the antiapoptotic Bcl-2 protein expression. These results suggest that adenovirus-mediated DLC1 transfer, alone or together with other agents such as inhibitors of Bcl-2, or histone deacetylases that were already tested in clinical trials might prove effective in the treatment of aggressive androgene independent and metastatic prostate cancer.

在过去的一年中，在了解DNA损伤在细胞衰老和衰老中的作用以及对肝癌1（DLC1）中删除的理解，基因抑制功能及其在治疗干预措施中的潜在用途方面取得了重大进展。 DNA损伤的积累不仅在衰老过程中，而且在诱导细胞衰老过程中起着至关重要的作用。细胞感测和修复DNA损伤随着年龄的增长而下降的能力。但是，这种依赖年龄依赖性下降的基本分子机制仍然难以捉摸。为了理解人类衰老过程中DNA损伤反应的定量和定性变化，磷酸化组蛋白H2AX（γ-H2AX）的性质和动态（特别发生在DNA双链断裂（DSB）的部位），在不同的患者的细胞中检查了不同的衰老患者的细胞中的细胞，并从癌症综合症中增加了癌症和癌症的相关性，并与癌症稳定相关。这项研究表明，γ-H2AX病灶的发生率通常随着供体年龄的增长而增加，尽管源自Werner综合征（WS）患者的细胞表现出的γ-H2AX病灶的发病率比来自类似年龄的正常捐助者的发病率要高得多。随着正常和WS成纤维细胞向衰老发展，培养物的进一步增加发生在培养中。 DSB修复蛋白募集到γ-H2AX灶的募集速率与正常供体和WS供体的年龄成反比，这可能部分归因于老年捐赠者伽马 - H2AX焦点的生长缓慢。由于基因组稳定性可能取决于DSB的有效处理，在这种情况下，γ-H2AX灶的快速形成以及DSB修复蛋白的快速积累可能具有损害性，因此我们的发现表明，这些过程中提高效率的提高可能导致基因组不稳定性与正常和病理衰老有关。 DLC1基因是由我们组孤立和特征的，最近已被独立证实为有效的善良肿瘤抑制基因。该基因的沉默或过度反应在几种常见的人类癌症（例如肝脏，乳腺癌，肺和前列腺癌）中起着重要作用，最重要的是，由DLC1调节的信号通路为治疗干预提供了广泛的靶标。凋亡的诱导是肿瘤抑制基因的主要属性。在对肺和前列腺肿瘤细胞的两项独立研究中，我们鉴定出表明DLC1诱导程序性细胞死亡的细胞和分子改变。 DLC1对体外细胞增殖具有巨大的抑制作用，并防止在裸鼠中接种人非小细胞肺癌细胞后体内肿瘤的发展。 DLC1在肺癌细胞中的转导总是诱导基因肿瘤抑制剂功能的细胞形态佐证的变化。 DLC1介导的基于肌动蛋白细胞骨架的形态学改变，而DLC1蛋白核转运发生在抑制细胞迁移之前和诱导凋亡之前。这些观察结果得出的结论是，DLC1在细胞质中起作用是肿瘤细胞增殖和迁移的抑制剂，但在细胞核中作为凋亡的诱导剂。根据我们的证据，DLC1现在被认为是转移抑制基因。肿瘤细胞从局部生长的原发性肿瘤传播到解剖远处的转移的过程是癌症死亡率的主要原因，占癌症伤亡的90％以上。转移性疾病是一个复杂的过程，涉及各种细胞和遗传改变。我们表明DLC1抑制了肝癌细胞向远处器官的扩散。肝癌细胞对远处器官的抑制与RhoA活性的降低，细胞骨架改变，骨桥脑蛋白（OPN）（OPN）和基质金属蛋白酶9（MMP9）的表达有关，这些表达在大多数具有相关转移酶的原发性肝肿瘤中都高度加大。 OPN和MMP-9的过度表达以及DLC1的沉默可能为肝癌预后提供预后标记。使用腺病毒载体作为基因转移的最有用的载体的DLC1基因治疗是预防HCC传播的现实前景。同样，寻找上调DLC-1基因表达的饮食成分和药理学剂可能会导致可能有助于预防转移有用的药物，这是肝癌进展过程中最有害的事件。 In prostate cancer, we found that recurrent downregulation and inactivation of DLC1 by epigenetic mechanisms may serve as an marker for early detection of prostate cancer and subsequently we showed that adenovirus-mediated DLC1 transfer in androgen-independent aggressive and metastatic prostate tumor cells inhibited cell proliferation in vitro, their tumorigenicity in nude mice, and induced凋亡。它还诱导细胞周期停滞，抑制RhoA的激活和肌动蛋白应激纤维的形成。在对组蛋白脱乙酰基酶抑制剂诱导的细胞死亡具有抗性的前列腺肿瘤细胞中，仅在化学抑制抗凋亡BCl-2蛋白表达后，DLC1表达的恢复才能诱导凋亡。这些结果表明，腺病毒介导的DLC1转移单独或与其他药物（例如Bcl-2的抑制剂）或已在临床试验中已经进行过测试的组蛋白脱乙酰基酶可能有效治疗侵袭性雄性雄激素独立和转移性前列腺癌。

项目成果

期刊论文数量（27）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

DNA variants of DLC-1, a candidate tumor suppressor gene in human hepatocellular carcinoma.

DOI：
10.3892/ijo.23.1.133
发表时间：
2003-07
期刊：
International journal of oncology
影响因子：
5.2
作者：
Sang-won Park;M. Durkin;S. Thorgeirsson;N. Popescu
通讯作者：
Sang-won Park;M. Durkin;S. Thorgeirsson;N. Popescu

Identification and characterization of a gene encoding a putative mouse Rho GTPase activating protein gene 8, Arhgap8.

编码假定的小鼠 Rho GTPase 激活蛋白基因 8（Arhgap8）的基因的鉴定和表征。

DOI：
10.1016/s0378-1119(02)01143-5
发表时间：
2003
期刊：
Gene
影响因子：
3.5
作者：
Shan,Zhihong;Haaf,Thomas;Popescu,NicholasC
通讯作者：
Popescu,NicholasC

DLC1 suppresses distant dissemination of human hepatocellular carcinoma cells in nude mice through reduction of RhoA GTPase activity, actin cytoskeletal disruption and down-regulation of genes involved in metastasis.

DOI：
10.3892/ijo_32_6_1285
发表时间：
1992-06
期刊：
International journal of oncology
影响因子：
5.2
作者：
Xiaoling Zhou;D. Zimonjic;Sang-won Park;Xuyu Yang;M. Durkin;N. Popescu
通讯作者：
Xiaoling Zhou;D. Zimonjic;Sang-won Park;Xuyu Yang;M. Durkin;N. Popescu

SMAD5 gene expression, rearrangements, copy number, and amplification at fragile site FRA5C in human hepatocellular carcinoma.

人肝细胞癌中 SMAD5 基因表达、重排、拷贝数和脆弱位点 FRA5C 的扩增。

DOI：
10.1016/s1476-5586(03)80041-6
发表时间：
2003
期刊：
Neoplasia (New York, N.Y.)
影响因子：
0
作者：
Zimonjic,DrazenB;Durkin,MarianE;Keck-Waggoner,CatherineL;Park,Sang-Won;Thorgeirsson,SnorriS;Popescu,NicholasC
通讯作者：
Popescu,NicholasC

Fragile sites and cancer genes on the short arm of chromosome 8.

8 号染色体短臂上的脆弱位点和癌症基因。