目的：探讨舒尼替尼通过NF-κB信号通路诱导肝癌HepG2细胞表达自然杀伤细胞2族成员D配体（natural killer group 2 member D ligands, NKG2DLs)的分子机制。方法： 常规体外培养HepG2细胞，单细胞凝胶电泳检测1 μmol/L舒尼替尼处理HepG2细胞24 h前后DNA损伤情况,实时荧光定量PCR检测药物处理前后细胞DNA损伤修复分子mRNA的表达，Western blotting 检测分别以NF-κB激动剂和抑制剂处理HepG2细胞前后NKG2DLs蛋白表达及IKKα和IκBα表达情况。结果： 舒尼替尼药物处理后，HepG2细胞均发生不同程度DNA损伤；且AP-1、ATM、ATR mRNA表达水平明显升高，而CHK1、CHK2、GSK3β mRNA表达水平明显降低；不同处理组间DNA损伤修复相关信号分子mRNA表达有显著差异（F=61.242，P=0.000）。NF-κB转录活性抑制剂JSH-23可降低HepG2细胞NKG2DLs蛋白表达量，而NF-κB转录活性激动剂TNF-α、PMA均可增加HepG2细胞NKG2DLs蛋白表达量（F=15.043，P=0000）；舒尼替尼处理肿瘤细胞后NF-κB的抑制分子IKKα被抑制，而激活分子IκBα被激活。结论： 舒尼替尼可通过DNA损伤修复分子激活NF-κB旁路途径诱导肿瘤细胞表达NKG2DLs。

Objective:To investigate molecular mechanism of sunitinib-induced expressions of natural killer group 2 member D ligands (NKG2DLs) in human hepatocellular carcinoma cell HepG2.Methods: HepG2 cells were cultivated by routine method. DNA damage in HepG2 cells was detected by single cell gel electrophoresis (SCGE) assay. The expressions of DNA damage-related molecule were quantitated by RT-qPCR. The levels of NKG2DLs, IKKα and IkBα were determined by immunoblotting. Results: Single cell gel electrophoresis revealed that HepG2 cells have various degree of DNA damages after exposed to sunitinib. RT-qPCR analysis showed that the expressions of AP-1, ATM, and ATR mRNA in HepG2 cells treated with sunitinib were significantly increased, whereas the levels of CHK1，CHK2，GSK3β mRNA were markedly lower. While the NF-κB inhibitor JSH-23 decreased the expressions of NKG2DLs in HepG2 cells, the agonist of NF-κB increased the expressions of these molecules. Furthermore, in HepG2 treated with sunitinib, IKKα was phosphorylated, and IkBα was activated. Conclusion: The data indicated that sunitinib induces the upregulated expressions of NKG2DLs in carcinoma cells by DNA damage-related molecules with activate the NF-κB pathway.

国家自然科学青年基金资助项目（No. 81302372,81300431）；南方医科大学珠江医院优秀中青年人才资助项目(No.201207008)