Suppression of Salivary Gland Apoptosis by AKt

描述：(申请人提供)诱导细胞凋亡是一项重要的 哺乳动物发育中的事件；然而，唾液腺细胞的凋亡 由头颈部照射、化疗药物或自身免疫引起的 疾病最终会降低这些患者的生活质量。目标是 这个项目的目的是定义唾液中激活的信号通路 并确定这些事件是否可以通过不同的凋亡刺激 被压制了。特异靶1识别抗细胞凋亡的激活 生长因子对C5腮腺和C6颌下细胞蛋白激酶Akt的影响 LINES和AKT在细胞凋亡损伤后激活的保护作用 (依托泊苷、紫杉醇、照射和FasL)。之后将对细胞凋亡进行量化 用流式细胞仪检测每种刺激因子的亚组分、半胱氨酸天冬氨酸蛋白酶3 Akt底物的活化、TUNEL试验和免疫印迹。转基因小鼠 已经被产生来表达一种符合宪法规定的支持生存的形式 唾液腺中的Akt分子(Myr-Akt)。《特定目标2》将评估 Myr-Akt小鼠原代唾液腺腺泡细胞的能力 抑制细胞凋亡。特异靶3检查细胞的凋亡反应 转基因小鼠对各种刺激包括：异丙肾上腺素、辐射和 法斯勒。每次刺激后，将测量唾液腺流率和 唾液腺组织将通过标准组织学、TUNEL染色、 Akt底物的免疫印迹和唾液蛋白的变化 浓度。这些研究将为我们更好地理解 并有可能为临床治疗由癌症引起的 唾液腺中的细胞凋亡。

DESCRIPTION: (provided by applicant) The induction of apoptosis is an important event in mammalian development; however, apoptosis of salivary gland cells resulting from head and neck irradiation, chemotherapeutic drugs, or autoimmune diseases ultimately decreases the quality of life for these patients. The goal of this project is to define the signaling pathways activated in the salivary glands by various apoptotic stimuli and determine if these events can be suppressed. Specific aim 1 identifies the activation of the anti-apoptotic protein kinase Akt by growth factors in C5 parotid and C6 submandibular cell lines and the protection activated Akt confers after an apoptotic insult (etoposide, taxol, irradiation, and FasL). Apoptosis will be quantitated after each stimulus by flow cytometry to measure Sub G1 fraction, caspase 3 activation, TUNEL assay, and immunoblotting for Akt substrates. Transgenic mice have been generated to express a constitutively active form of the pro-survival molecule Akt (Myr-Akt) in the salivary glands. Specific aim 2 will assess the ability of the primary murine salivary gland acinar cells from the Myr-Akt mice to suppress apoptosis. Specific aim 3 examines the apoptotic response of the transgenic mice to various stimuli including: isoproterenol, irradiation, and FasL. After each stimulus, salivary gland flow rate will be measured and salivary gland tissues will be analyzed by standard histology, TUNEL staining, and immunoblotting for Akt substrates and changes in salivary protein concentrations. These studies will provide greater insight to the understanding of this tissue and potentially provide clinical treatment of cancer-induced apoptosis in salivary glands.