TM4SF1通过受体型酪氨酸激酶DDR1调节胰腺癌侵袭转移的分子机制研究

Pancreatic carcinoma is a kind of malignant tumor with very poor prognosis. One of the major hallmarks of pancreatic cancer is its early systemic dissemination and its extraordinary local tumor progression. In our previous experiments, the results showed that TM4SF1 was highly expressed in pancreatic cancer tissue and almost all pancreatic cancer cell lins.The migration and invasion of pancreatic cancer cell lines were significantly decreased when TM4SF1 was silenced in comparison with their respective controls. In contrast, TM4SF1 silencing had no effect on cell proliferation.The result of IP showed discoidin domain receptor 1（DDR1） was combined with TM4SF1. Therefore, our hypothesis is TM4SF1 promotes pancreatic cancer migration and invasion, and TM4SF1 effected through DDR1. Our study is aimed to detcect the mRNA and protein expression of TM4SF1 in clinical samples so that we could analyze the correlation between the expression of TM4SF1 and clinical characteristics especially invasion and metastasis. The invasion and matastasis of pancreatic cancer will be investigated after TM4SF1 silenced in vitro and in the orthotopic pancreatic cacer in nude mice. We will detect the expression and activities of DDR1 after TM4SF1 silenced.We will prove the combination of TM4SF1 and DDR1 by cell transfection and pull-down assay. The migtation and invasion will be investgated when TM4SF1 silenced as well as DDR1 activated, so that we can prove TM4SF1 was combined DDR1 to promte the migration and invasion in pancreatic cancer. The aim is to provide new ideas and experimental basis for research of invasion and metastasis in pancreatic cancer.

胰腺癌的高侵袭性及早期发生转移是胰腺癌恶性程度高、预后差的重要原因。前期实验发现，四次跨膜蛋白L6家族成员TM4SF1在胰腺癌组织及多数胰腺癌细胞株中高表达。体外实验提示TM4SF1与胰腺癌细胞的迁移、侵袭相关，而与细胞增殖无明显相关。免疫共沉淀提示TM4SF1与盘状结构域受体酪氨酸激酶1（DDR1）相互作用。因此，我们认为TM4SF1促进胰腺癌的侵袭与转移，并且可能通过DDR1发挥作用。本研究旨在通过对临床标本的检测，分析TM4SF1与临床指标尤其是侵袭与转移的相关性；并通过体内、体外实验观察TM4SF1对胰腺癌细胞的影响；观察TM4SF1与DDR1表达及活化的关系；应用细胞转染及pull-down技术进一步证实TM4SF1与DDR1的相互作用；沉默TM4SF1同时过表达DDR1后是否可以恢复细胞的迁移、侵袭能力。我们的课题为胰腺癌侵袭与转移的研究提供了新思路和实验基础。

胰腺癌的高侵袭性及早期发生转移是胰腺癌恶性程度高、预后差的重要原因。前期实验发现，四次跨膜蛋白L6家族成员TM4SF1在胰腺癌组织及多数胰腺癌细胞株中高表达。体外实验提示TM4SF1与胰腺癌细胞的迁移、侵袭相关，而与细胞增殖无明显相关。免疫共沉淀提示TM4SF1与盘状结构域受体酪氨酸激酶1（DDR1）相互作用。因此，我们认为TM4SF1促进胰腺癌的侵袭与转移，并且可能通过DDR1发挥作用。本研究旨在通过对临床标本的检测，分析TM4SF1与临床指标尤其是侵袭与转移的相关性；并通过体内、体外实验观察TM4SF1对胰腺癌细胞的影响；观察TM4SF1与DDR1表达及活化的关系；应用细胞转染及pull-down技术进一步证实TM4SF1与DDR1的相互作用；沉默TM4SF1同时过表达DDR1后是否可以恢复细胞的迁移、侵袭能力。在实验研究中，课题组发现TM4SF1在胰腺癌组织和细胞系中高表达，并且与吉西他滨耐药及肿瘤血管形成相关。TM4SF1通过DDR1调节MMP2和MMP9的表达，降解细胞外基质，从而介导癌细胞的迁移、侵袭。课题组并依据实验结果，撰写并发表了2篇直接相关的SCI文章、2篇相关SCI文章、2篇核心期刊和1篇会议论文。为进一步探索胰腺癌侵袭、转移提供新思路和实验基础。