超声靶向微泡破坏介导JNK信号通路对肝癌淋巴道转移作用和机制的研究

The most important difference between malignant and benign tumors is metastasis. To epithelial malignant tumors, the early metastatic mode is lymphatic metastasis which is an important factor which affects prognosis of the patients. Therefore, it is very important to investigate the molecular mechanism of lymphatic metastasis and find new approaches of intervention. Primary hepatocellular carcinoma (HCC) is one of the most common malignant tumor with high incidence of a disease in our country. Although a mass of researches have been done, its mechanism is not clear. Hca-P and Hca-F is a pair of syngenetic mouse hepatocarcinoma ascites cell lines which has the different rates of lymphatic metastasis. Hca-F is the cell lines whose metastatic rate is more than 70% and Hca-P less than 30%. They are the ideal models for the researches of mechanism of lymphatic metastasis . Our experimental group has engaged in the research work for the mechanism of lymphatic metastasis. We have screened out the lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines by using suppressive subtractive hybridization and gene chip assays respectively and obtained the lymphatic metastasis-associated proteins by using quantitative proteomics technique. The expressing level of JNK was much higher in Hca-F than that in Hca-P cell lines in both gene and protein levels which showed that JNK1 maybe play an important role in lymphatic metastasis of mouse hepatocarcinoma. The aim of this study is to investigate the different biochemical behaviors of Hca-P and Hca-F cell lines by different expression levels of JNK1 by down and up regulations mediated by ultrasound microbubbles. Therefore, we can understand the influence on proliferation, migration and invasion of mouse hepatocellular carcinoma Hca-P and Hca-F celllines after up-regulation and down-regulation of of JNK1 expression. We probably know the key target gene point of HCC. In the same time, the safety and effectiveness of ultrasound microbubbles will be researched. It will provide us a theoretical basis for inhibition treatment for lymphatic metastasis of HCC.

上皮来源恶性肿瘤的早期转移方式是淋巴道转移，其发生机制是肿瘤学面临的难题。本研究已采用基因芯片、定量蛋白质组学等技术，对来源于同一亲本细胞、具有高低淋巴道转移能力的小鼠肝癌细胞株中筛选出的基因JNK1进行研究。在前期工作的基础上，拟应用超声靶向微泡破坏技术（UTMD）介导基因转染分别上调及下调JNK1在高及低淋巴道转移力小鼠肝癌细胞株中的表达，以细胞增殖、粘附、迁移、侵袭等体外实验及动物体内实验对其上调及下调后生物学行为的变化进行研究。应用UTMD技术联合JNK1-shRNA抑制肝癌淋巴道转移。同时观察JNK信号通路下游的重要基因CD44v6和paxillin，与淋巴结转移相关的基因VEGF-C和VEGF-D的变化。通过上述工作对JNK信号通路可能促进肝癌淋巴道转移的作用和机制进行深入研究，为肝癌淋巴道转移的治疗寻找新靶点，为超声微泡介入肝癌淋巴道转移的基因靶向治疗提供理论依据和新的方法。

上皮来源恶性肿瘤的早期转移方式是淋巴道转移，其发生机制是肿瘤学面临的难题。本研究已采用基因芯片、定量蛋白质组学等技术，对来源于同一亲本细胞、具有高低淋巴道转移能力的小鼠肝癌细胞株中筛选出的基因JNK1进行研究。在前期工作的基础上，拟应用超声靶向微泡破坏技术（UTMD）介导基因转染上调JNK1在低淋巴道转移力小鼠肝癌细胞株（Hca-P细胞）中的表达，以细胞增殖、粘附、迁移、侵袭等体外实验及动物体内实验对其上调后生物学行为的变化进行研究。同时观察JNK信号通路下游的重要基因CD44v6和paxillin，与淋巴结转移相关的基因VEGF-C和VEGF-D的变化。通过上述工作对JNK信号通路可能促进肝癌淋巴道转移的作用和机制进行深入研究，为肝癌淋巴道转移的治疗寻找新靶点，为超声微泡介入肝癌淋巴道转移的基因靶向治疗提供理论依据和新的方法。