胞内氯离子通道蛋白1（CLIC1）在肺腺癌侵袭转移中的作用及机制

Earlier invasion and metastasis are the most significant biological behaviors of lung adenocarcinoma. Recently，it was proved that intracellular chloride channel protein 1 ( CLIC1 ) plays important roles in both physiological procedures and tumor genesis. However，whether CLIC1 is associated with lung adenocarcinoma is not yet reported. In our previous works, we primarily found that intracellular chloride channel protein 1 ( CLIC1 ) is obvious over-expression in lung adenocarcinoma, and it is correlated with both the T stage and the prognosis. We also found that the invasion and the metastasis of lung adenocarcinoma will be obviously depressed when the CLIC1 expression was depressed. We hypothesize that CLIC1 and its protein-protein interaction network play important roles in the invasion and metastasis of lung adenocarcinoma. In this research, we will detect the role of CLIC1 in lung adenocarcinoma invasion and metastasis by using over-expression and knocked-down CLIC1 in vitro and vivo respectively.Then, SILAC technique in combination with LTQ-Orbitrap MS is used to explore the CLIC1 interacting proteins. Through the above research, we want to further elucidate the mechanism of lung adenocarcinoma invasion metastasis,and to find new molecular markers for anti-invasion and anti- metastasis of lung adenocarcinoma.

较早出现侵袭转移是肺腺癌最显著的生物学特征，也是导致患者死亡的主要原因。胞内氯离子通道1（CLIC1)近年来被发现在肿瘤的发生发展中起着重要作用。但目前有关CLIC1与肺腺癌的关系尚无报道。前期研究中，我们首次发现CLIC1在肺腺癌中过表达，与肿瘤的T分期及预后相关，在肺腺癌细胞中CLIC1被沉默后，细胞的侵袭转移能力明显下降。我们推测，CLIC1及其上下游的相互作用蛋白网络在肺腺癌的侵袭转移中起着重要的作用。本课题拟首先通过抑制和过表达CLIC1后在体、内外检测其在肺腺癌侵袭转移中的作用机制；再利用稳定同位素标记细胞培养（SILAC）和质谱技术筛选出与肺腺癌侵袭转移相关的CLIC1相互作用蛋白，并在体内、外检测这些蛋白在肺腺癌侵袭转移中的作用机制。通过上述研究，旨在进一步阐明肺腺癌侵袭转移的分子机制，为肺腺癌发生侵袭转移的预测提供新的分子标记物，为肺腺癌的抗侵袭转移提供新的分子靶标。

较早出现侵袭转移是肺腺癌最显著的生物学特征，也是导致患者死亡的主要原因。胞内氯离子通道1（CLIC1)近年来被发现在肿瘤的发生发展中起着重要作用。但目前有关CLIC1与肺腺癌的关系尚无报道。前期研究中，我们首次发现CLIC1在肺腺癌中过表达，与肿瘤的T分期及预后相关，在肺腺癌细胞中CLIC1被沉默后，细胞的侵袭转移能力明显下降。我们推测，CLIC1及其上下游的相互作用蛋白网络在肺腺癌的侵袭转移中起着重要的作用。.1）通过构建细胞株，细胞实验、裸鼠皮下接种实验，证明了CLIC1对肿瘤具有抑癌作用，主要调节肿瘤的侵袭力。.2）通过蛋白组学及转录组学测序获得一批CLIC1影响的下游基因列表，筛选并确定与CLIC1相互作用且与肺腺癌侵袭转移密切相关的关键蛋白及其网络： .3）生物信息学分析CLIC1作用网络中的关键蛋白,进一步证明CLIC1作用的关键通路，阐明这些关键蛋白DTYMK在肺腺癌侵袭和转移中的作用及其分子机制，为肺腺癌转移复发和预后的预测提供分子靶标。.4）通过分子对接虚拟计算获得靶向CLIC1靶点的候选小分子化合物，为下一步的临床应用研究打下了工作基础。