Progression of Metastasis of Oral Tongue Cancer

描述（由申请人提供）：目前的肿瘤进展理论假设正常上皮细胞需要获得逃避细胞凋亡的机制，并获得其他一些必要的生物学特性，才能进展为侵袭性和转移性肿瘤。细胞脱离基膜后的程序性死亡被称为细胞脱落。本应用的中心假设是，漏诊与局部肿瘤进展和口腔鳞状细胞癌（SCCOC）的转移有关。四个相互关联的假设将在四个单独的目标中进行评估。第一种假设是，抗嗜酸药对肿瘤进展是必要的，但不是充分的，包括局部生长以及区域和远处转移。为了证明抗气味剂与疾病进展相关，我们将在我们开发的原位裸鼠模型中研究人类SCCOC细胞系的生长。体外选择的人类SCCOC细胞系和anoikis抗性细胞将被注射到裸鼠的舌头中，并检查这些小鼠的肿瘤生长，存活时间以及区域淋巴结和远处转移的存在。相反，我们将通过对悬浮培养中细胞存活率的定量分析，证明我们从局部和远处转移的体内原位选择中获得的细胞系对anoikis的抗性。指导本应用的下一个目标的第二个假设是，介导anoikis抗性的促生存信号不是组成的，而是由迄今为止的细胞脱离过程诱导的，这表明这是由于诱导下游的细胞凋亡抑制，因为我们已经发现细胞脱离通过外在和内在途径诱导对多种形式的细胞凋亡诱导的抵抗。第三种假说认为，BiR中含有的蛋白c-IAP-2、survivin和XIAP在面对多种形式的凋亡信号传导时介导了脱离诱导的存活。这一假设将通过在5个细胞系中增加或减少这两种调节分子的表达或活性来验证，并检验由此产生的细胞系在体外的抗氧化性以及在体内的致瘤性和转移潜力。最后一个假设是，下游凋亡调节分子c-IAP-2、survivin和XIAP的表达与人类SCCOC和口腔癌原位模型的肿瘤进展和转移有关。为了证明这些凋亡调节分子在人类SCCOT肿瘤中表达，并且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来评估存档的人类SCCOT肿瘤中这些凋亡调节分子的表达。预计在临床病理结果较差的患者的肿瘤标本中，凋亡抑制分子的高表达将为IAPsas的预后指标和/或治疗靶点的潜在作用提供见解。在SCCOT原位转移模型中，这些分子的表达增加导致肿瘤生长和转移更具侵袭性，这将进一步支持IAPs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.