目的：探讨舒尼替尼诱导人肝癌HepG2细胞表达的自然杀伤细胞2族成员D配体（natural killer group 2 member D ligands，NKG2DLs）与核因子kappa B（nuclear factor B kappa, NF-κB）信号通路之间的相互作用。方法：常规体外培养HepG2细胞，利用实时荧光定量PCR检测1 μmol/L舒尼替尼处理HepG2细胞24 h前后NF-κB基因家族成员 mRNA的表达情况，利用计算机辅助设计并合成NF-κB1、NF-κB2和RelB siRNA引物，通过脂质体法将目的基因siRNA转染于HepG2细胞，荧光显微镜观察转染情况，实时荧光定量PCR检测干扰效率，Western blotting检测siRNA转染前后HeG2细胞内NF-κB1、NF-κB2和RelB蛋白表达，流式细胞术检测siRNA前后HepG2细胞NKG2DLs表达率。结果： 实时荧光定量PCR结果显示，舒尼替尼处理HepG2细胞后，NF-κB1、NF-κB2和RelB mRNA表达水平升高，主要以NF-κB2 mRNA和RelB mRNA升高为主。荧光显微镜观测siRNA转染后HepG2细胞红色荧光表达率约为60%，干扰效率达95%以上。siRNA转染后HepG2细胞内NF-κB1、NF-κB2和RelB蛋白表达水平明显下降，siRNA转染+药物组NKG2DLs表达率明显低于药物处理组（P<0.05）。结论： 首次揭示了舒尼替尼通过NF-κB旁路途径诱导肿瘤细胞表达NKG2DLs。

Objective: To investigate the interaction between the natural killer group 2 member D ligands (NKG2DLs) expressions and NF-κB signaling pathway in HepG2 cells treated with sunitinib.Methods: HepG2 cells were cultivated by routine method. The mRNAs of NF-κB gene family members in HepG2 cells were quantitated by RT-qPCR. The siRNAs specific for NF-κB1, NF-κB2 and RelB were designed using a computer soft and synthesized. They were transfected into HepG2 cells using liposome. The transfection and interference efficacy were evaluated by fluorescence microscopy, and real-time quantification PCR respectively. The expressions of NF-κB1, NF-κB2 and RelB were determined by immunoblotting, and the expressions of NKG2DLs were assessed by Flow cytometry.Results: The mRNAs of NF-κB1, and especially NF-κB2 and RelB were significantly increased in HepG2 cells treated with sunitinib. In siRNA transfection experiments, the red fluorescence was present in about 60% of HepG2 cells under fluorescence microscope, and the interference efficiency was 95%. Immunoblotting revealed that NF-κB1, NF-κB2 and RelB were decreased significantly in HepG2 cells after the transfection. Flow cytometry analysis found that the expressions of NKG2DLs in the siRNA transfected group was significantly lower than that in drug treatment group (P<005). Conclusion: Sunitinib induces the expressions of NKG2DLs on hepatocellular carcinoma HepG2 cells by the activating the alternative 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项目资助（No.201207008)