MODULATION OF CELL-CELL COMMUNICATION IN BREAST CANCER

细胞之间通过缝隙连接的直接通信代表着 向形成高度分化的方向迈出的重要一步 多细胞生物。在转化或癌变的细胞中有一种 细胞间通讯异常。缝隙接头的重新安装 通过调制剂进行的通信已被演示在 包括我们在内的调查人员的细胞。使用二甲基亚砜和环状 在体外治疗中，我们的实验室能够将非传染性， 荷尔蒙非依赖性小鼠乳腺肿瘤细胞对肿瘤的逆转作用 他莫昔芬治疗和变得可传染，如染料所示 转移技术。这项建议的具体目的是延长 在以下三个领域进行了实验： (1)间隙结的电导已被证明是由 有几个因素，其中之一是细胞内钙离子水平。这个 建议的研究是为了调节人类细胞间的通讯 影响细胞内钙离子浓度的药物对乳腺癌细胞的作用 通过三个独立但相互作用的系统，即循环-AMP， 钙调蛋白和蛋白激酶C系统，以及(B)通过使用 抗缝隙连接蛋白的单抗。路西法黄 刮载、微量注射和荧光法转移染料 光漂白后再分配(FRAP)技术将用于 确定主要测压元件和测压元件之间的传输速率 相邻细胞，细胞内钙离子水平将是 被监视着。 (2)也有越来越多的人认识到间质 乳腺成分在生长发育中起着重要的作用 对乳腺癌的监管。因此，传播学也将 在乳腺癌细胞和正常间质细胞之间进行实验，以及 乳腺癌和转化的间质细胞之间的关系 共文化体系。调节剂对这种相互作用的影响 并对胶原酶和透明质酸的产量进行了评估。 (3)这项提案还将研究雌激素之间的相互作用。 受体(ER)阳性和阴性乳腺癌细胞，并评估 这种调节将改变这两个细胞群体的生长平衡 在共同文化体系中。 通过改变细胞间的交流程度和细胞行为， 这些调节剂可能会延缓肿瘤细胞的生长，最终导致 可以延长乳腺癌患者的生存期。

Direct communication between cells via gap junctions represents an important evolutionary step towards formation of highly differentiated multicellular organisms. In transformed or cancerous cells there is an aberration of intercellular communication. Re-installment of gap junctional communication by modulating agents has been demonstrated in transformed cells by investigators including us. Using dimethyl sulfoxide and cyclic AMP in vitro treatment, our laboratory was able to convert noncommunicable, hormone-independent mouse mammary tumor cells to tumors which regressed on tamoxifen therapy and become communicable as illustrated by the dye transfer technique. The specific aims of this proposal are to extend the experiments to the following three fields: (1) The conductance of the gap junction has been shown to be gated by several factors, one of which is the intracellular calcium ion level. The proposed studies are to modulate the cell-cell communication of human breast cancer cells (a) by agents which affect the intracellular calcium level through three separate but interacted systems, namely the cyclic-AMP, the calmodulin and the protein kinase C systems, and (b) by the use of monoclonal antibodies against the gap junctional proteins. Lucifer yellow dye transfer by the scrape-load, the microinjection and the fluorescence redistribution after photobleaching (FRAP) techniques will be used to determine the rate of transfer between the primary loading cells and neighboring cells, and the intracellular calcium ion level will be monitored. (2) There is also growing appreciation of the concept that the stromal component of the mammary gland plays an important role in the growth regulation of breast cancer. Therefore, the communication study will also be performed between breast cancer cells and normal stromal cells, and between breast cancer and transformed stromal cells in an in vitro coculture system. The influence of modulating agents on such interaction and on the collagenase and hyaluronate production will be assessed. (3) This proposal will also examine the interaction between estrogen receptor (ER) positive and negative breast cancer cells and assess whether such modulation will alter the growth balance of these two cell populations in the coculture system. By changing the degree of cell-cell communication and cellular behavior, these modulating agents may retard tumor cell growth and ultimately result in a prolongation of survival in patients with breast cancer.