Progression of Metastasis of Oral Tongue Cancer

描述(申请人提供)：目前的肿瘤进展理论假设，正常的上皮细胞需要获得逃避凋亡的机制，并获得其他几个基本的生物学特性，才能进展为侵袭性和转移性肿瘤。细胞从基底膜脱离后的程序性死亡称为Oiki。这一应用的中心假设是，逃避失巢凋亡与局部肿瘤进展和口腔鳞癌(SCCOC)转移有关。四个独立的相互关联的假设将在四个不同的目标中进行评估。第一个假设是，失巢细胞抵抗对于肿瘤的进展是必要的，但不是充分的，包括局部生长以及区域和远处转移。为了证明失巢凋亡抵抗与疾病进展有关，我们将在我们建立的原位裸鼠模型中研究人类SCCOC细胞系的生长。体外筛选的人SCCOC细胞株和抗失巢蛋白的细胞将被注射到裸鼠的舌内，并将检查这些小鼠的肿瘤生长、存活时间以及是否存在区域结节和远处转移。相反，我们将通过量化细胞系在悬浮培养中的存活率来证明我们通过在体内原位选择区域和远处转移而获得的细胞系对失巢凋亡的抗性。指导本应用下一个目的的第二个假设是，介导失巢凋亡抵抗的亲生存信号不是结构性的，而是由迄今为止的细胞分离数据过程诱导的，这表明这是由于远在下游的凋亡抑制的诱导，因为我们已经发现细胞分离诱导对多种形式的凋亡诱导的抵抗，通过外在和内在的途径。第三种假设认为，在多种形式的凋亡信号面前，含有c-IAP-2、Survivin和XIAP蛋白的BIR介导了脱离诱导的生存。这一假设将通过增加和减少这两个调节分子中的任何一个在5个细胞系中的表达或活性，并检查所产生的细胞系在体外对失巢凋亡的抵抗力以及它们在体内的致瘤性和转移潜力来检验。最后一个假设是，下游的凋亡调节分子c-IAP-2、Survivin和XIAP的表达与人SCCOC和口腔癌原位模型中的肿瘤进展和转移有关。为了证明这些凋亡调节分子在人SCCOT肿瘤中的表达，以及它们的表达与较差的临床病理结果有关，我们将通过原位杂交和免疫组织化学的方法来评估这些凋亡调节分子在人SCCOT肿瘤中的表达。临床病理转归差的患者肿瘤标本中高表达的凋亡抑制分子有望为研究IAPs作为预后指标和/或治疗靶点的潜在作用提供依据。在SCCOT的原位转移模型中，这些分子的表达增加会导致更具侵袭性的肿瘤生长和转移，这将进一步支持IAP的这些作用。

DESCRIPTION (provided by applicant): Current theories of tumor progression postulate that normal epithelial cells need to acquire mechanisms to evade apoptosis and acquire several other essential biologic properties in order to progress to invasive and metastatic tumors. Programmed cell death after detachment from the basement membrane is known as an oikis. The central hypothesis is of this application is that evasion of anoikis is associated with local tumor progression, and metastasis of squamous cell carcinoma of the oral cavity (SCCOC). Four individual inter-related hypotheses would be evaluated in four separate Aims. The first hypothesis is that anoikis-resistance is necessary but not sufficient for tumor progression, including local growth as well as regional and distant metastases. To demonstrate that anoikis-resistance is associated with disease progression, we will investigate the growth of human SCCOC cell lines in an orthotopic, nude mouse model that we have developed. Human SCCOC cell lines and anoikis resistant cells that have been selected in vitro will be injected into the tongues of nude mice, and these mice will be examined for tumor growth, length of survival and the presence of regional nodal and distant metastases. Conversely, we will demonstrate the anoikis resistance of cell lines that we have derived from in vivo orthotopic selection of regional and distant metastases by quantitation of their survival in suspension culture. The second hypothesis guiding the next Aim of this application is that the pro-survival signals mediating anoikis resistance are not constitutive but rather are induced by the process of cell detachment data to date, suggest that this is due to the induction of apoptosis suppression that is far downstream, as we have found that cell detachment induces resistance to multiple forms of apoptosis induction by both the extrinsic and intrinsic pathways. The third hypothesis is that the BiR containing proteins c-IAP-2, survivin, and XIAP are mediating the detachment-induced survival in the face of multiple forms of apoptotic signaling. This hypothesis will be tested by increasing and decreasing expression or activity of either of these two regulatory molecules in5cell lines and examining the resultant cell lines for their anoikis resistance in vitro and their tumorigenicity and metastatic potential in vivo. The final hypothesis is that expression of the downstream apoptotic regulatory molecules c-IAP-2, survivin, and XIAP is associated with tumor progression and metastasis in human SCCOC and in an orthotopic model of oral cancer. To demonstrate that these apoptotic regulatory molecules are expressed in human SCCOT tumors and that their expression is linked with poorer clinic pathologic outcomes, archival human SCCOT tumor will be evaluated for expression of these apoptotic regulatory molecules by in situ hybridization and immunhistochemistry. It is anticipated that high expression of apoptotic inhibitory molecules in tumor specimens from patients with poor clinicopathologic outcomes will provide insight into the potential roles of the IAPsas prognostic indicators and/or therapeutic targets. Further support for these roles of IAPs will result from demonstration that increased expression of these molecules leads to more aggressive tumor growth and metastasis in an orthotopic metastatic model of SCCOT.