钙释放激活钙通道crac对乳腺癌细胞增殖的影响及机制生理学专业论文.docxVIP

中文摘要中文摘要 中文摘要 中文摘要 背景：cE+控制着众多的细胞功能，钙池操纵的Ca：+内流(Store。Operated Calcium Entry,SOCE)参与控制许多重要的Ca2十信号生理过程，SOCE也以电流 形式被记录为钙释放激活钙电流(Ca2+release．activated Ca2+，CRAC)。已有研究 证明CRAC通道是由位于细胞膜上的Orai和位于内质网膜上的stromal interaction molecule(STIM)两种蛋白构成，并在肥大细胞、T淋巴细胞、内皮细胞、平滑 肌等多种细胞中表达，最近一项研究表明CRAC通道参与乳腺癌细胞迁移与转 移，抑制或沉默CRAC通道，乳腺癌细胞的迁移能力和转移灶的形成受阻。 目的：探明CRAC与恶性乳腺癌细胞增殖之间的关系，为肿瘤治疗提供新的 药物靶点。 方法：本实验以小鼠EMT6细胞和人MDA．MB．231细胞两种乳腺癌细胞系为 研究对象，应用Western blotting和RT-PCR方法分别从蛋白水平和mRNA水平检测 STIMl和Orail在肿瘤细胞中的存在性；显微镜下形态学观察以及细胞增殖检测 实验观察细胞经CRAC通道抑制剂．2．APB处理后的增殖情况：应用Western blotting、流式细胞术、基因沉默技术探讨CRAC通道与乳腺癌细胞增殖相关的内 在机制。 结果：两种乳腺癌细胞系上存在由STIMl和Orail组成的CRAC通道；细胞形 态学和增殖实验证明2-APB浓度越高，乳腺癌细胞增殖受阻现象越明显； 1009M2．APB处理48h能够显著降低细胞中STIMl和Orail的表达，乳腺癌细胞大 部分停滞在G0／Gl期、S期细胞比例减少，基因沉默CRAC通道后细胞增殖受阻现 象明显。 关键词：CRAC 乳腺癌 2．APB Orai STn订 AbstractAbstract Abstract Abstract Background：Store—operated calcium entry(SOCE)is essential for Ca2+-mediated physiological processes and functions．SOCE call also be recorded as electrical current named Ca2+release activated Ca2+current(CRAC)．Previous studies established that two classes of proteins，stromal interaction molecule(STIM)and Oral， constitute CRAC channels．and these channel components have been identified in many cell types such as mast cells，T lymphocytes，endothelial cells，vascular smooth muscle cells，etc．A recent study demonstrated the presence of CRAC in breast CanCer cells where it Was associated with cell migration and metastasis． Objective：To study the role of CRAC channel in Pdlncer eell proliferation and to provide new therapeutic strategy intervening the malignancy． Methods：We focused on two breast cancer eell lines，mouse EMT6 and human MDA．MB．23 1．Wbtern blotting and IU’-PCR were used to detect the expression of STIM 1 and Orai l．Cell proliferation was measured by observing cell morphology and cell counting kits．2-aminoethoxydiphenyl borate(2-APB)，a CRAC channel blocker, and siRNA technique were used to inhibit function of CRAC channel．Flow cytometry Was employed to quantitate th