ACTIVATION SPECIFIC C ERBB 2 DETECTION IN BREAST CANCER

大约50%的乳腺癌患者有淋巴 结节阴性肿瘤。尽管这类患者的总体预后是 显著好于那些有正向结节的人，30%的人仍然 死于转移性疾病。结节阳性疾病的治疗体会 这表明这个假阴性的患者队列可能会衍生出显著的 生存受益于辅助性系统治疗。高-高的识别 因此，风险结节阴性的患者是一个临床高度重视的领域。 利息。 C-erbB-2(neu，HER-2)受体的表达被认为是一种 预测人类乳腺癌自然病程的标准，但 这种受体在早期疾病中的预测性是一个问题 争论很多。我和我的同事最近的观察可能有助于确定 C-erbB-2的真正致病和预后意义 表型。我们已经证明c-erbB-2受体可能存在于 可相互转换的形式--一种激活的175 kDa酪氨酸磷酸化形式 (P175)和无活性的185 kDa酪氨酸去磷酸化形式(P185)。这个 这两种形式的存在(这两种形式在传统上是无法区分的 C-erb B-2检测技术)提供了可能的解释 上面提到的临床争论。我建议澄清一下临床上 C-erbB-2基因在乳腺癌组织中的表达及意义 并使用一种新的实验方法量化体内受体的激活 接近。该项目有三个目标： 第一个目标是制备抗p175和p185的多克隆抗血清 利用含有起始激活和失活的c-erbB-2多肽 分别选址； 第二个目标是产生针对完整的 P175和p185分子通过表达突变型全长人c- 杆状病毒中的ErbB-2基因； 第三个目标是确定p175和p175的相对表达。 P185在正常、侵袭前和浸润性乳腺组织中的表达，从而 确定这些c-erb B-2的生物学和预后意义 人类乳腺癌中的受体亚型。 考虑到我目前的临床情况，这些目标尤其可行 与研究所内的外科和病理部门保持联系， 用于同位素的超灵敏荧光成像仪的实验室可用性 研究表明，无限量供应高亲和力磷酸酪氨酸抗体 受体活性控制，以及我和我的同事的经验 在生长因子/酪氨酸激酶领域。实现这些目标 目标将为更雄心勃勃的拨款申请铺平道路 我建议评估抗p175和/或抗p185的临床价值 一项对接受辅助系统治疗的患者进行的前瞻性研究 用于结节阴性的乳腺癌。

Approximately 50% of patients presenting with breast cancer have lymph node-negative tumors. Although the overall prognosis for such patients is substantially better than for those with positive nodes, 30% will still succumb to metastatic disease. Experience with node-positive disease suggests that this 'false-negative' patient cohort may derive significant survival benefit from adjuvant systemic therapy. Identification of high- risk node-negative patients is therefore an area of intense clinical interest. Expression of the c-erbB-2 (neu, HER-2) receptor has been proposed as one criterion for predicting the natural history of human breast cancer, but the predictivity of this receptor in early-stage disease is an issue of much debate. Recent observations by my colleagues and I may help establish the true pathogenetic and prognostic significance of the c-erbB-2 phenotype. We have shown that c-erbB-2 receptors may exist in two interconvertible forms - an activated 175kDa tyrosine-phosphorylated form (p175) and an inactive 185kDa tyrosine-dephosphorylated form (p185). The existence of these two forms (which are indistinguishable by conventional c-erbB-2 detection techniques) provides a possible explanation for the clinical polemic mentioned above. I propose to clarify the clinical significance of c-erbB-2 expression in human breast cancer by identifying and quantifying receptor activation in vivo using a novel experimental approach. The project has three objectives: The first objective is to prepare polyclonal antisera against p175 and p185 using c-erbB-2 peptides containing the prime activation and inactivation sites respectively; The second objective is to generate monoclonal antibodies to the intact p175 and p185 molecules obtained by expressing mutant full-length human c- erbB-2 cDNAs in baculovirus; The third objective is to determine the relative expression of p175 and p185 in normal, preinvasive and invasive breast tissue, and thus to establish the biologic and prognostic significance of these c-erbB-2 receptor isoforms in human breast cancer. These objectives are particularly feasible given my current clinical liaison with surgical and pathology services within the Institute, the laboratory availability of an ultrasensitive PhosphorImager for isotope studies, an unlimited supply of high-affinity phosphotyrosine antibody for receptor activity controls, and the experience of myself and my colleagues in the growth factor/tyrosine kinase field. Achievement of these objectives will prepare the way for a more ambitious grant application in which I propose assessing the clinical value of anti-p175 and/or anti-p185 in a prospective study of patients undergoing adjuvant systemic treatment for node-negative breast cancer.