舒尼替尼上调耐药鼻咽癌细胞NKG2D配体表达增强NK细胞抗肿瘤作用的分子机制研究

As for the drug-resistant relapse nasopharyngeal carcinoma (NPC)，Combining usage of sunitinib malate and NK cells immunotherapy may result in synergistic antitumor effect. The effect reached is based on the sunitinib up-regulate NKG2DL ligands on tumor cells, resulting in enhanced immune eradication of NK cells. But Its is unknown which the molecular mechanism of sunitinib possessed inducing effects on NKG2DL ligands expression of tumor cells. The previous research proves that sunitinib treatment activated expressions of signaling transduction molecules and apoptosis molecule correlated with DNA damage-repair response and cell apoptosis, at as same time also induced NF-κB expression on tumor cells. According to these results, it is speculated that NF-κB gene may be a key gene for up-regulate NKG2DL ligands on tumor cells. In the thesis, we will research interaction relationship with DNA damage-repair response including AP-1/ATR/CHK1/CHK2 and cell apoptosis including P53/Bax/PUMA/NOXA and function of NF-κB by useing molecular biological and immunological technology on the drug-resistant relapse nasopharyngeal carcinoma cells with or without sunitinib treatment. In the following, we will research molecular path that NF-κB gene could be up-regulated NKG2D ligands expression of tumor cells. We will first discovered mechanisms of sunitinib inducing effects of NKG2D ligands on tumor cells. Moreover, we provide theoretical and experimental bases for establishment of effective regiment against drug-resistant NPC with combining usage of molecular targeted agents and NK cells immunotherapy.

舒尼替尼联合NK细胞治疗耐药鼻咽癌可实现协同抗肿瘤作用，其依据是药物上调靶细胞NKG2D配体的表达，增强NK细胞杀伤活性，但药物上调靶细胞表达NKG2D配体的分子机制仍未清楚。前期研究发现舒尼替尼作用肿瘤细胞后，阻断靶细胞特异性靶点，活化DNA损伤修复分子及细胞凋亡分子，导致NF-κB高表达，因此，我们推测NF-κB可能是上调靶细胞表达NKG2D配体的关键基因。项目拟利用分子生物学和免疫学技术在体内外研究舒尼替尼处理鼻咽癌细胞前后，靶细胞DNA损伤修复分子（AP-1/ATR/CHK1/CHK2）、凋亡分子（P53、Bax、PUMA、NOXA）与NF-κB基因之间的相互作用关系，以及上调靶细胞表达NKG2D配体的分子路径，以阐明舒尼替尼上调靶细胞表达NKG2D配体的分子机制，为分子靶向药物联合NK细胞治疗耐药鼻咽癌提供理论依据。

舒尼替尼联合NK细胞治疗耐药肿瘤可实现协同抗肿瘤作用，其依据是药物上调靶细胞NKG2D配体（natural killer group 2 member D ligand NKG2DL）的表达，增强NK细胞杀伤活性，但药物上调靶细胞表达NKG2D配体的分子机制仍未清楚。我们推测NF-κB可能是上调靶细胞表达NKG2D配体的关键基因。项目拟利用分子生物学和免疫学技术在体内外研究舒尼替尼处理鼻咽癌细胞前后，靶细胞DNA损伤修复分子（AP-1/ATR/CHK1/CHK2）、凋亡分子（P53、Bax、PUMA、NOXA）与NF-κB基因之间的相互作用关系，以及上调靶细胞表达NKG2D配体的分子路径，以阐明舒尼替尼上调靶细胞表达NKG2D配体的分子机制。我们研究结果发现，一方面，舒尼替尼能阻断肿瘤细胞增殖和血管生成信号通路促进细胞凋亡，另一方面，舒尼替尼诱导肿瘤细胞表达NK细胞活化性配体NKG2DL，促进肿瘤细胞对NK细胞杀伤敏感性。舒尼替尼诱导肿瘤细胞表达NKG2DL，主要通过DNA损伤修复反应分子和细胞凋亡通路与转录因子NF-κB之间相互作用，活化由NF-κB2和RelB组成的旁路途径调节NKG2DL的转录和表达。舒尼替尼通过NF-κB旁路途径诱导肿瘤表达NKG2D配体的分子机制为多靶点酪氨酸激酶抑制剂联合NK细胞治疗肿瘤提供实验理论依据。