MECHANISMS OF TUMOR CELL INVASION

肿瘤细胞通过基底膜和间质的侵袭 组织允许肿瘤转移并在体内新的位置定居。 然而，细胞侵袭的调节方式是完全 未知。最近的研究表明，酪氨酸蛋白的磷酸化 膜结合蛋白在卵巢癌转移侵袭中起着至关重要的作用 细胞。这些研究表明，酪氨酸激酶及其膜结合 底物高度升高与不定足有关， 侵袭细胞前沿的细胞突起。形成 金雀异黄素可阻断内翻足细胞和随后的细胞入侵。 酪氨酸激酶的抑制剂。附着体和脱落的膜泡 人恶性黑色素瘤细胞系LOX及其膜泡 从实验模型中生长的LOX中提取的是丰富的来源 与入侵相关的抗原。制备抗原的起始原料是 已可用于分离侵袭相关的酪氨酸磷酸化 蛋白质和几种酪氨酸磷酸化蛋白已被鉴定 在使用现有抗体的分离的介形虫膜组分中。这 拟议的工作将：1)分离和微测序主要的未鉴定的 食虫草和落膜中的酪氨酸磷酸化多肽 LOX细胞的囊泡和LOX肿瘤的膜 磷酸酪氨酸单抗，以及2)识别现有的和 产生新的抗这些侵袭相关的单抗 酪氨酸磷酸化蛋白。蛋白质测序与单抗 在这里获得的抗体将使我们能够识别酪氨酸磷酸化 仅定位于侵袭性褶皱的蛋白质，其 与不定足类的关联可能形成了对照的分子基础 肿瘤细胞的侵袭和转移。

Invasion of tumor cells through basement membranes and interstitial tissues allows tumors to metastasize and colonize new sites in the body. However, the manner in which cell invasion is regulated is completely unknown. Recent studies indicate that tyrosine phosphorylation of membrane-bound proteins is critically important in metastatic invasion of cells. These studies show that tyrosine kinases and their membrane-bound substrates are highly elevated in association with invadopodia, the cellular protrusions at the leading edge of invasive cells. Formation of invadopodia and subsequent cell invasion were blocked by genistein, an inhibitor of tyrosine kinases. Invadopodia and shed membrane vesicles of the human malignant melanoma cell line LOX, as well as membrane vesicles derived from LOX grown in experimental models are a rich source of invasion-related antigens. Starting material for antigen preparation is already available to isolate invasion related-tyrosine phosphorylated proteins and several tyrosine phosphorylated proteins have been identified in isolated invadopodia membrane fractions using existing antibodies. This proposed work will: 1) isolate and microsequence the major unidentified tyrosine phosphorylated polypeptides from invadopodia and shed membrane vesicles of LOX cells, and membranes of LOX tumors using anti- phosphotyrosine monoclonal antibodies, and 2) identify existing and generate new monoclonal antibodies against these invasion-associated tyrosine phosphorylated proteins. Protein sequencing and monoclonal antibodies obtained here will allow us to identity tyrosine phosphorylated proteins that localize exclusively to invasive ruffles and whose association with invadopodia may form the molecular basis for the control of tumor cell invasion and metastasis.